Sample records for proline accumulation parameter

  1. Salt stress encourages proline accumulation by regulating proline biosynthesis and degradation in Jerusalem artichoke plantlets.

    Huang, Zengrong; Zhao, Long; Chen, Dandan; Liang, Mingxiang; Liu, Zhaopu; Shao, Hongbo; Long, Xiaohua


    Proline accumulation is an important mechanism for osmotic regulation under salt stress. In this study, we evaluated proline accumulation profiles in roots, stems and leaves of Jerusalem artichoke (Helianthus tuberosus L.) plantlets under NaCl stress. We also examined HtP5CS, HtOAT and HtPDH enzyme activities and gene expression patterns of putative HtP5CS1, HtP5CS2, HtOAT, HtPDH1, and HtPDH2 genes. The objective of our study was to characterize the proline regulation mechanisms of Jerusalem artichoke, a moderately salt tolerant species, under NaCl stress. Jerusalem artichoke plantlets were observed to accumulate proline in roots, stems and leaves during salt stress. HtP5CS enzyme activities were increased under NaCl stress, while HtOAT and HtPDH activities generally repressed. Transcript levels of HtP5CS2 increased while transcript levels of HtOAT, HtPDH1 and HtPDH2 generally decreased in response to NaCl stress. Our results supports that for Jerusalem artichoke, proline synthesis under salt stress is mainly through the Glu pathway, and HtP5CS2 is predominant in this process while HtOAT plays a less important role. Both HtPDH genes may function in proline degradation.

  2. Effects of Salt Stress on Proline Accumulation in Relation to Osmotic Adjustment in the Seedling of Elymus Dahuricus Turcz



    We investigated the effects of salt stress on proline accumulation in the seedling of Elymus dahuricus Turcz. from two different geographic regions in order to explicit various adaptability in response to salt stress condition. Plants were dealt with in five salt treatments (0%, 0.5%, 1%, 2%, 3%). Significant differences were obvious at the highest salinity treatments for almost each considered parameters. Given that proline concentration is closely related to salt tolerance, we found that G2 seemed to be more tolerant. Furthermore, the proline concentration might be exploited as a dependable measure for evaluating the effect of considering regional factors on salt tolerance.

  3. Hydrophytes lack potential to exhibit cadmium stress induced enhancement in lipid peroxidation and accumulation of proline

    Dhir, Bhupinder; Sharmila, P.; Saradhi, P. Pardha


    Investigations were carried out to evaluate if hydrophytes (viz. Ceratophyllum, Wolffia, and Hydrilla) can be used as markers to assess the level of heavy metal pollution in aquatic bodies. The potential of these hydrophytes for lipid peroxidation and accumulation of proline in response to cadmium (Cd{sup 2+}) pollution was studied. Hydrophytes were raised in artificial pond water (APW) supplemented with various levels of Cd{sup 2+}. Interestingly, unlike mesophytes none of the hydrophytes showed ability to accumulate proline. Infact, in response to Cd{sup 2+} pollution hydrophytes exhibited a decline in proline levels in comparison to controls but mesophytes (viz. Brassica juncea, Vigna radiata and Triticum aestivum) showed progressive increase in the level of proline with increase in the extent of Cd{sup 2+} pollution. Mesophytes showed six to nine-fold increase in the level of proline in response to 1 mM Cd{sup 2+}. The potential of the above hydrophytes for lipid peroxidation was also low under Cd{sup 2+} stress. In contrast, as expected a significant enhancement in the lipid peroxidation was observed in all three mesophytes in response to their exposure to Cd{sup 2+}. About two-fold increase in production of malondialdehyde (a cytotoxic product of lipid peroxidation) was recorded in mesophytes exposed to 1 mM Cd{sup 2+}. However, a decline in chlorophyll (Chl a and Chl b) levels was recorded in response to Cd{sup 2+}pollution both in hydrophytes as well as mesophytes. In summary, hydrophytes neither have potential to accumulate proline nor have ability to accelerate lipid peroxidation under heavy metal stress. This suggests that the adaptive mechanism(s) existing in hydrophytes to tackle heavy metal stress is distinct from that in mesophytes.

  4. Free proline accumulation in leaves of cultivated plant species under water deficit conditions

    Hanna Bandurska


    Full Text Available The effect of water deficit caused by soil drought on the content of free proline as well as the degree of cell membrane damages in the leaves of three cultivated plant species having different farm usefulness and water requirements have been studied. The used pIants were: poinsettia (Euphorbia pulcherrima Willd., 'Regina' and 'Cortez' grown for decorative purposes, a green vegetable of broccoli (Brassica oleracea var. botrytis, subvar. cymosa, 'Colonel' and 'Marathon' and a cereal plant of barley (the wild form Hordeum spontaneumm and Hordeum vulgaree 'Maresi'. The examined species differed in the size of the experienced stress. the Iargest RWC reduction was found iii broccoli leaves, while somewhat smaller - in barley. In poinsettia leaves, the reduction of RWC level was not large or did not occur at all. The accumulation of free proline in the species under study was also variable. The largest amount of this amino acid tended to accumulate in broccoli leaves, whereas the increase of its level took place only at a strong dehydration of tissues. The increase of proline level was smaller in barley leaves than in broccoli, but that was found already at a smalI dehydration of tissues. In poinsettia leaves, a several f`old increase of proline level was found at the early stage of the stress. The level of that amino acid gradually increased at consecutive times and did not depend on tissue dehydration. Damage of cell membranes amounted to 8.5-9.5% in barley leaves, about 3% in brocolli and to 0-2.6% in poinsettia. The role of proline in prevention of leaf dehydration and in alleviation of dehydration effects in the studied species has been discussed.

  5. Effect of salt stress on growth, Na+ accumulation and proline metabolism in potato (Solanum tuberosum) cultivars.

    Jaarsma, Rinse; de Vries, Rozemarijn S M; de Boer, Albertus H


    Potato (Solanum tuberosum) is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na(+) homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na(+) levels in root and stem with the highest leaf Na(+) concentration of all cultivars, resulting in a high Na(+) shoot distribution index (SDI) for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na(+) accumulation was found and the SDI for Na(+) points to a role of stem Na(+) accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH) expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na(+) accumulation in stem tissue, resulting in reduced Na(+) transport to the leaves.

  6. Effect of salt stress on growth, Na+ accumulation and proline metabolism in potato (Solanum tuberosum cultivars.

    Rinse Jaarsma

    Full Text Available Potato (Solanum tuberosum is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na(+ homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na(+ levels in root and stem with the highest leaf Na(+ concentration of all cultivars, resulting in a high Na(+ shoot distribution index (SDI for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na(+ accumulation was found and the SDI for Na(+ points to a role of stem Na(+ accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1 gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na(+ accumulation in stem tissue, resulting in reduced Na(+ transport to the leaves.

  7. Effect of Salt Stress on Growth, Na+ Accumulation and Proline Metabolism in Potato (Solanum tuberosum) Cultivars

    Jaarsma, Rinse; de Vries, Rozemarijn S. M.; de Boer, Albertus H.


    Potato (Solanum tuberosum) is a major crop world-wide and the productivity of currently used cultivars is strongly reduced at high soil salt levels. We compared the response of six potato cultivars to increased root NaCl concentrations. Cuttings were grown hydroponically and treated with 0 mM, 60 mM and 180 mM NaCl for one week. Growth reduction on salt was strongest for the cultivars Mozart and Mona Lisa with a severe senescence response at 180 mM NaCl and Mozart barely survived the treatment. The cultivars Desiree and Russett Burbank were more tolerant showing no senescence after salt treatment. A clear difference in Na+ homeostasis was observed between sensitive and tolerant cultivars. The salt sensitive cultivar Mozart combined low Na+ levels in root and stem with the highest leaf Na+ concentration of all cultivars, resulting in a high Na+ shoot distribution index (SDI) for Mozart as compared to Desiree. Overall, a positive correlation between salt tolerance and stem Na+ accumulation was found and the SDI for Na+ points to a role of stem Na+ accumulation in tolerance. In stem tissue, Mozart accumulated more H2O2 and less proline compared to the tolerant cultivars. Analysis of the expression of proline biosynthesis genes in Mozart and Desiree showed a clear reduction in proline dehydrogenase (PDH) expression in both cultivars and an increase in pyrroline-5-carboxylate synthetase 1 (P5CS1) gene expression in Desiree, but not in Mozart. Taken together, current day commercial cultivars show promising differences in salt tolerance and the results suggest that mechanisms of tolerance reside in the capacity of Na+ accumulation in stem tissue, resulting in reduced Na+ transport to the leaves. PMID:23533673

  8. Temperature stress in accumulation of free proline of pigeonpea seedlings from seeds treated with polyamines

    Jéssica da Silva


    Full Text Available It was studied the effects of seed treatment with polyamines of pigeonpea for proline content of seedling in order to verify that the application of these polyamines attenuates temperature stress on germination and early seedling growth, and see if exogenous putrescine and spermidine induce the accumulation of proline, this amino acid may be biochemical and physiological indicator in seedlings that are under suboptimal temperatures and supraoptmail. The seeds of pigeonpea cv. BRS Mandarin and cv. Caqui, treated with a solution of 0.5 mM of putrescine and spermidine, were subjected to suboptimal temperature (20ºC, 18ºC, 16ºC and 14ºC and supraoptimal (36ºC, 38ºC, 40ºC and 44ºC for 24 hours and 48 hours. After these periods, were subjected to 25°C until day 10, when the percentage of seed germination were observed, the proline content of vegetative parts and the dry mass of the seedlings. A completely randomized design in a factorial arrangement was used 3x4x2+3 (solutions with polyamines, temperature stress by cooling/heating, time of exposure to stress, plus additional factors em25ºC with 0.0 mM and 0.5mM of Put and Spd, with four replicates of 25 seeds. The results were submitted to analysis of variance and means were compared by Tukey test at 5% probability, separately for each cultivar. Both exogenous polyamines attenuate the effects of cooling and heating, contributing to the growth of germinated seeds. Also, putrescine and spermidine exogenous mitigated the adverse effects by inducing proline accumulation, which leads to osmotic adjustment temperatures, although this physiological response has not minimized the negative effects of temperature stress on seedling growth of pigeonpea. Proline can be considered biochemical and physiological indicator in seedlings of both cultivars of pigeonpea treated with polyamines under temperature stress.

  9. Effects of Formulated Fertilizer Synergist on Abscisic Acid Accumulation, Proline Content and Photosynthetic Characteristics of Rice under Drought

    WANG Shao-xian; XIA Shi-tou; PENG Ke-qin; KUANG Feng-chun; CAO Yong; XIAO Lang-tao


    To investigate the effects of formulated fertilizer synergist on the drought tolerance in rice, pot experiment was conducted to analyze the photosynthetic characteristics and the accumulation of abscisic acid (ABA) and proline in middle-season rice variety Peiliangyou 93. The synergist could improve the net photosynthetic rate, and coordination between the water loss and the CO2 absorption as well as reduce the harmful effect on photosynthetic process under drought conditions. Under drought, the ABA accumulated massively both in roots and leaves, while the ABA content in roots was far higher than that in leaves. The results indicate that synergist could increase the ABA accumulation, but reduce the proline accumulation in rice plant under drought.

  10. Exogenous ornithine is an effective precursor and the δ-ornithine amino transferase pathway contributes to proline accumulation under high N recycling in salt-stressed cashew leaves.

    da Rocha, Iza Marineves Almeida; Vitorello, Victor Alexandre; Silva, Jamille Santos; Ferreira-Silva, Sérgio Luiz; Viégas, Ricardo Almeida; Silva, Evandro Nascimento; Silveira, Joaquim Albenisio Gomes


    The role of the δ-ornithine amino transferase (OAT) pathway in proline synthesis is still controversial and was assessed in leaves of cashew plants subjected to salinity. The activities of enzymes and the concentrations of metabolites involved in proline synthesis were examined in parallel with the capacity of exogenous ornithine and glutamate to induce proline accumulation. Proline accumulation was best correlated with OAT activity, which increased 4-fold and was paralleled by NADH oxidation coupled to the activities of OAT and Δ(1)-pyrroline-5-carboxylate reductase (P5CR), demonstrating the potential of proline synthesis via OAT/P5C. Overall, the activities of GS, GOGAT and aminating GDH remained practically unchanged under salinity. The activity of P5CR did not respond to NaCl whereas Δ(1)-pyrroline-5-carboxylate dehydrogenase was sharply repressed by salinity. We suggest that if the export of P5C from the mitochondria to the cytosol is possible, its subsequent conversion to proline by P5CR may be important. In a time-course experiment, proline accumulation was associated with disturbances in amino acid metabolism as indicated by large increases in the concentrations of ammonia, free amino acids, glutamine, arginine and ornithine. Conversely, glutamate concentrations increased moderately and only within the first 24h. Exogenous feeding of ornithine as a precursor was very effective in inducing proline accumulation in intact plants and leaf discs, in which proline concentrations were several times higher than glutamate-fed or salt-treated plants. Our data suggest that proline accumulation might be a consequence of salt-induced increase in N recycling, resulting in increased levels of ornithine and other metabolites involved with proline synthesis and OAT activity. Under these metabolic circumstances the OAT pathway might contribute significantly to proline accumulation in salt-stressed cashew leaves. Copyright © 2011 Elsevier GmbH. All rights reserved.

  11. Effect of salinity on seed germination, accumulation of proline and free amino acid in Pennisetum glaucum (L.) R. Br.

    Sneha, Sonam; Rishi, Anirudha; Dadhich, Amit; Chandra, Subhash


    Salinity is a major threat to agriculture, plants exhibits a variety of responses to salt stress that enable them to tolerate and survive in such conditions. Salinity affects physiological and biochemical processes in plants. A short term salt stress induced physiological and biochemical response were observed in P. glaucum. The experiment was conducted to understand the influence of salinity on seed germination, proline and free amino acid accumulation in P. glaucum. It was observed that as the salt concentration increased the germination percentage decreased as compared to control as well as the root/shoot length also decreased. This suggests that salinity greatly influences the germination as well as the plant growth. The levels ofbiochemical components proline and free amino acid were measured during the salt stressed condition. The 14 days old seedlings were subjected to 4 salt treatments (50, 100, 150 and 200 mM NaCI), free proline and free amino acids was calculated at 0, 12, 24, 48, 72 and 96th hour. Proline and free amino acid content in the salt stressed tissues increased with increase in salt concentration as well as with duration of salt stress. This result suggests that proline and free amino acid acids acts as compatible solutes in P. glaucum to protect the cellular macromolecules, maintain the osmotic balance and also scavenge the free radicals under salt stressed condition.

  12. Proline accumulation, ions dynamics and sodium root-shoot partition and compartmentation

    Jesus Emanuel eBojorquez Quintal


    Full Text Available Despite its economic relevance, little is known about salt tolerance mechanisms in pepper plants. To address this question, we compared differences in responses to NaCl in two Capsicum chinense varieties: Rex (tolerant and Chichen-Itza (sensitive. Under salt stress (150 mM NaCl over 7 days roots of Rex variety accumulated 50 times more compatible solutes such as proline compared to Chichen-Itza. Mineral analysis indicated that Na+ is restricted to roots by preventing its transport to leaves. Fluorescence analysis suggested an efficient Na+ compartmentalization in vacuole-like structures and in small intracellular compartments in roots of Rex variety. At the same time, Na+ in Chichen-Itza plants was compartmentalized in the apoplast, suggesting substantial Na+ extrusion. Rex variety was found to retain more K+ in its roots under salt stress according to a mineral analysis and microelectrode ion flux estimation (MIFE. Vanadate-sensitive H+ efflux was higher in Chichen-Itza variety plants, suggesting a higher activity of the plasma membrane H+-ATPase, which fuels the extrusion of Na+, and, possibly, also the re-uptake of K+. Our results suggest a combination of stress tolerance mechanisms, in order to alleviate the salt-induced injury. Furthermore, Na+ extrusion to apoplast does not appear to be an efficient strategy for salt tolerance in pepper plants.

  13. Putrescine catabolism via DAO contributes to proline and GABA accumulation in roots of lupine seedlings growing under salt stress

    Jolanta Legocka


    Full Text Available The levels of polyamines (PAs, proline (Pro, and γ-aminobutyric acid (GABA as well as the activity of diamine oxidase (DAO; EC were studied in the roots of 2-day-old lupine (Lupinus luteus L. ‘Juno’ seedlings treated with 200 mM NaCl for 24 h. The effect of adding 1 mM aminoguanidine (AG, an inhibitor of DAO activity, was also analyzed. It was found that in roots of lupine seedlings growing under salt stress, a negative correlation between Pro accumulation and putrescine (Put content takes place. Pro level increased in roots by about 160% and, at the same time, Put content decreased by about 60%, as a result of ca. twofold increase of DAO activity. The AG added to the seedlings almost totally inhibited the activity of DAO, increased Put accumulation to control level, decreased Pro content by about 25%, and reduced GABA level by about 22%. Addition of 50 mM GABA to the lupine seedlings growing in the presence of AG and NaCl restored Pro content in roots to its level in NaCl-treated plants. In this research, the clear correlation between Put degradation and GABA and Pro accumulation was shown for the first time in the roots of seedlings growing under salt stress. This could be considered as a short-term response of a plant to high salt concentration. Our findings indicate that during intensive Pro accumulation in roots induced by salt stress, the pool of this amino acid is indirectly supported by GABA production as a result of Put degradation.

  14. Response of the accumulation of proline in the bodies of Populus euphratica to the change of groundwater level at the lower reaches of Tarim River

    CHEN Yaning; CHEN Yapeng; LI Weihong; ZHANG Hongfeng


    The content of proline in the plant bodies is closely related to the converse-succession-resistant capability of the plants. In this paper, the relationship between the proline accumulation in the bodies of Populus euphratica and the change of groundwater level is analyzed by taking Populus euphratica, the main community-building species of the desert riparian forests along the Tarim River, as the research object. The research results show that the accumulation of proline in the bodies of Populus euphratica is closely related to the change of groundwater level gradient under drought stress, it increases with the drawdown of groundwater level and the increase of moisture stress degree; the accumulation of proline in the bodies of Populus euphratica has two extremely high points at the groundwater depth ranges of 3.64-5.14 m and 9.46-10.16 m. Combining the field investigation and the analysis of the plots, it is considered that the groundwater level of 3.5-4.5 m is rational for the growth of Populus euphratica. The stress groundwater depth for the normal growth and the critical one for the survival of Populus euphratica are below 4.5 m and 9-10 m respectively at the lower reaches of the Tarim River.

  15. Proline accumulation in leaves of Periploca sepium via both biosynthesis up-regulation and transport during recovery from severe drought.

    Yuyan An

    Full Text Available Drought resistance and recovery ability are two important requisites for plant adaptation to drought environments. Proline (Pro metabolism has been a major concern in plant drought tolerance. However, roles of Pro metabolism in plant recovery ability from severe drought stress are largely unexplored. Periploca sepium Bunge has gained increasing attention for its adaptation to dry environments. Here, we investigated Pro metabolism in different tissues of P. sepium seedlings in the course of drought stress and recovery. We found that leaf Pro metabolism response during post-drought recovery was dependant on drought severity. Pro biosynthesis was down-regulated during recovery from -0.4 MPa but increased continually and notably during recovery from -1.0 MPa. Significant correlation between Pro concentration and Δ1-pyrroline-5-carboxylate synthetase activity indicates that Glutamate pathway is the predominant synthesis route during both drought and re-watering periods. Ornithine δ-aminotransferase activity was up-regulated significantly only during recovery from -1.0 MPa, suggesting positive contribution of ornithine pathway to improving plant recovery capacity from severe drought. In addition to up-regulation of biosynthesis, Pro transport from stems and roots also contributed to high Pro accumulation in leaves and new buds during recovery from -1.0 MPa, as indicated by the combined analysis of Pro concentration and its biosynthesis in stems, roots and new buds. Except its known roles as energy, carbon and nitrogen sources for plant rapid recovery, significant positive correlation between Pro concentration and total antioxidant activity indicates that Pro accumulation can also promote plant damage repair ability by up-regulating antioxidant activity during recovery from severe drought stress.

  16. Effects of mannitol induced osmotic stress on proline accumulation, pigment degradation, photosynthetic abilities and growth characters in C3 rice and C4 sorghum

    Suriyan CHA-UM; Souvanh THADAVONG; Chalermpol KIRDMANEE


    Osmotic stress is one of the most important abiotic factors which inhibit growth and development in both the vegetative and reproductive stages of many plant species. The aim of this investigation was to compare the biochemical and physiological responses in C3 rice and C4 sorghum to water deficit. Chlorophyll a (Chla), chlorophyll b (Chlb), total chlorophyll (TC) and total carotenoid (Cx+c) contents in both rice and sorghum seedlings under osmotic stress were adversely affected, related to increasing osmotic pressure in the culture media. In addition, the chlorophyll's fluorescence parameters and net photosynthetic rate (Pn) decreased, leading to growth reduction. Also, a positive correlation was found between physiological and biochemical data, while proline accumulation showed a negative relationship. The Chlb, Pn and fresh weight were maintained better in osmotic-stressed (-1.205 MPa) C4 sorghum seedlings than those in C3 rice seedlings. The growth and physiological responses of C3 rice and C4 sorghum decreased depending on the plant species, the osmotic pressure in the media and their interactions. Pigment content and Pn ability in C4 sorghum grown under mannitol-induced osmotic stress increased to a greater degree than in C3 rice, resulting in maintenance of growth.

  17. Effect of exogenous γ-aminobutyric acid treatment on proline accumulation and chilling injury in peach fruit after long-term cold storage.

    Shang, Haitao; Cao, Shifeng; Yang, Zhenfeng; Cai, Yuting; Zheng, Yonghua


    The effect of exogenous γ-aminobutyric acid (GABA) on chilling injury of peach fruit was investigated. Freshly harvested peaches were treated with 1, 5, or 10 mM GABA at 20 °C for 10 min and then stored at 1 °C for up to 5 weeks. The results showed that all of the GABA treatments could reduce chilling injury of peach fruit with 5 mM being the most effective concentration. GABA treatment significantly enhanced the accumulation of endogenous GABA and proline, which resulted from the increased activities of glutamate decarboxylase, Δ(1)-pyrroline-5-carboxylate synthetase, and ornithine δ-aminotransferase and decreased proline dehydrogenase activity. Our results revealed that GABA treatment may be a useful technique to alleviate chilling injury in cold-stored peach fruit, and the reduction in chilling by GABA may be due to the induction of endogenous GABA and proline accumulation. These data are the first evidence that exogenous GABA induced chilling tolerance in postharvest horticultural products.

  18. Proline and soluble sugars accumulation in three pepper species (Capsicum spp) in response to water stress imposed at different stages of growth

    Gideon O Okunlola; Richard O Akinwale; Adekunle A Adelusi


    Drought is a major production constraint for major fruits and vegetable crops in the tropics. This study was conducted to in-vestigate the effect of limited water supply at three growth stages (vegetative, flowering and fruiting) on the accumulation of proline and soluble sugars in three pepper species. Seeds of the three pepper species,Capsicum chinense Jacq., C. annuumL. andC. frutescens L. were raised in a nursery and the seedlings were transplanted into seventy two plastic pots arranged in a randomized complete block design with three replicates, 25 days after planting. Four water treatments, 200 mL of water supplied twice daily (W1), once in every three days (W2), once in every five days (W3), and zero water supplied throughout growing period (W0) were imposed at three vegetative, flowering, and fruiting growth stages. Data were collected on relative water content, free proline and total soluble sugar. Data collected were subjected to analysis of variance and means were separated using Duncan's multiple range test. Results show that the concentration of proline and soluble sugar in leaves of the three pepper species were found to be remarkable at the different stages of growth in the stressed plants.

  19. AB-QTL analysis reveals new alleles associated to proline accumulation and leaf wilting under drought stress conditions in barley (Hordeum vulgare L.

    Sayed Mohammed A


    Full Text Available Abstract Background Land plants have evolved several measures to maintain their life against abiotic stresses. The accumulation of proline is the most generalized response of plants under drought, heat or salt stress conditions. It is known as an osmoprotectant which also acts as an instant source of energy during drought recovery process. But, both its role and genetic inheritance are poorly understood in agriculture crops. In the present work, advanced backcross quantitative trait locus (AB-QTL analysis was performed to elucidate genetic mechanisms controlling proline accumulation and leaf wilting in barley under drought stress conditions. Results The analysis revealed eight QTL associated to proline content (PC and leaf wilting (WS. QTL for PC were localized on chromosome 3H, 4H, 5H and 6H. The strongest QTL effect QPC.S42.5H was detected on chromosome 5H where drought inducible exotic allele was associated to increase PC by 54%. QTL effects QPC.S42.3H, QPC.S42.4H and QPC.S42.6H were responsible to heighten PC due to the preeminence of elite alleles over the exotic alleles which ranged from 26% to 43%. For WS, QTL have been localized on chromosome 1H, 2H, 3H and 4H. Among these, QWS.S42.1H and QWS.S42.4H were associated to decrease in WS due to the introgression of exotic alleles. In addition, two digenic epistatic interaction effects were detected for WS where the additive effect of exotic alleles imparted a favorable increase in the trait value. Conclusions The present data represents a first report on whole-genome mapping of proline accumulation and leaf wilting in barley. The detected QTL are linked to new alleles from both cultivated and wild accessions which bring out an initial insight on the genetic inheritance of PC and WS. These QTL alleles are fixed in the isogenic background of Scarlett, which will allow for positional cloning of underlying genes and to develop drought resilient barley cultivars.

  20. AB-QTL analysis reveals new alleles associated to proline accumulation and leaf wilting under drought stress conditions in barley (Hordeum vulgare L.).

    Sayed, Mohammed A; Schumann, Henrik; Pillen, Klaus; Naz, Ali A; Léon, Jens


    Land plants have evolved several measures to maintain their life against abiotic stresses. The accumulation of proline is the most generalized response of plants under drought, heat or salt stress conditions. It is known as an osmoprotectant which also acts as an instant source of energy during drought recovery process. But, both its role and genetic inheritance are poorly understood in agriculture crops. In the present work, advanced backcross quantitative trait locus (AB-QTL) analysis was performed to elucidate genetic mechanisms controlling proline accumulation and leaf wilting in barley under drought stress conditions. The analysis revealed eight QTL associated to proline content (PC) and leaf wilting (WS). QTL for PC were localized on chromosome 3H, 4H, 5H and 6H. The strongest QTL effect QPC.S42.5H was detected on chromosome 5H where drought inducible exotic allele was associated to increase PC by 54%. QTL effects QPC.S42.3H, QPC.S42.4H and QPC.S42.6H were responsible to heighten PC due to the preeminence of elite alleles over the exotic alleles which ranged from 26% to 43%. For WS, QTL have been localized on chromosome 1H, 2H, 3H and 4H. Among these, QWS.S42.1H and QWS.S42.4H were associated to decrease in WS due to the introgression of exotic alleles. In addition, two digenic epistatic interaction effects were detected for WS where the additive effect of exotic alleles imparted a favorable increase in the trait value. The present data represents a first report on whole-genome mapping of proline accumulation and leaf wilting in barley. The detected QTL are linked to new alleles from both cultivated and wild accessions which bring out an initial insight on the genetic inheritance of PC and WS. These QTL alleles are fixed in the isogenic background of Scarlett, which will allow for positional cloning of underlying genes and to develop drought resilient barley cultivars.

  1. The matrikine N-acetylated proline-glycine-proline induces premature senescence of nucleus pulposus cells via CXCR1-dependent ROS accumulation and DNA damage and reinforces the destructive effect of these cells on homeostasis of intervertebral discs.

    Feng, Chencheng; Zhang, Yang; Yang, Minghui; Lan, Minghong; Liu, Huan; Wang, Jian; Zhou, Yue; Huang, Bo


    Intervertebral disc (IVD) cell senescence is a recognized mechanism of intervertebral disc degeneration (IDD). Elucidating the molecular mechanisms underlying disc cell senescence will contribute to understanding the pathogenesis of IDD. We previously reported that N-acetylated proline-glycine-proline (N-Ac-PGP), a matrikine, is involved in the process of IDD. However, its roles in IDD are not well understood. Here, using rat nucleus pulposus (NP) cells, we found that N-Ac-PGP induced premature senescence of NP cells by binding to CXCR1. N-Ac-PGP induced DNA damage and reactive oxygen species accumulation in NP cells, which resulted in activation of the p53-p21-Rb and p16-Rb pathways. Moreover, the RT(2) profiler PCR array showed that N-Ac-PGP down-regulates the expression of antioxidant genes in NP cells, suggesting a decline in the antioxidants of NP cells. On the other hand, N-Ac-PGP up-regulated the expression of matrix catabolic genes and inflammatory genes in NP cells. Concomitantly, N-Ac-PGP reinforced the destructive effects of senescent NP cells on the homeostasis of the IVDs in vivo. Our study suggests that N-Ac-PGP plays critical roles in the pathogenesis of IDD through the induction of premature senescence of disc cells and via the activation of catabolic and inflammatory cascades in disc cells. N-Ac-PGP also deteriorates the redox environment of disc cells. Hence, N-Ac-PGP is a new potential therapeutic target for IDD.

  2. Effect of water withdrawal on formation of free radical, proline accumulation and activities of antioxidant enzymes in ZAT12-transformed transgenic tomato plants.

    Chandra Rai, Avinash; Singh, Major; Shah, Kavita


    Water stress often leads to the accumulation of reactive oxygen species (ROS) and their excessive production alters the activities of enzymes involved in their removal. ZAT12 is a member of stress-responsive C(2)H(2) type Zinc Finger Protein (ZFP) reported to control the expression of several stress-activated genes in plants through ROS signaling. The ZAT12-transformed tomato lines (cv. H-86 variety Kashi Vishesh) when subjected to water withdrawal for 7, 14 and 21 days revealed significant and consistent changes in activities of enzymes SOD, CAT, APX, GR and POD paralleled with an increased proline levels. Unlike that in wild-type tomato, the leaf superoxide anion and hydrogen peroxide concentrations in the transformed tomato plants did not alter much, suggesting a well regulated formation of free radicals suppressing oxidative stress in the latter. Results suggest BcZAT12-transformed tomato lines ZT1, ZT2 and ZT6 to be better adapted to drought stress tolerance by accumulation of osmolyte proline and increased antioxidant response triggered by the ZAT12 gene. Therefore, the ZAT12-transformed tomato cv. H-86 lines will prove useful for higher yield of tomato crop in regions affected with severe drought stress.

  3. Smoke Priming, a Potent Protective Agent Against Salinity: Effect on Proline Accumulation, Elemental Uptake, Pigmental Attributes and Protein Banding Patterns of Rice (Oryza Sativa

    Jamil, Muhammad


    Full Text Available The exogenous application of plant derived smoke solution through seed pre treatment is consider to create tolerance in the plant against salinity, for this purpose different dilution of plant derived smoke solution as 1:5000 Buhania, 1:1000 Buhania, 1:1000 Cymbopogon, 1:500 Cymbopogon were used against 0 mM, 50, 100 and 150mM NaCl solution in the medium. The effect was observed on total proline accumulation, heavy metals uptake, photosynthetic pigments and protein polypeptide bands intensity in two rice varieties as Basmati 385 (B-385 and Shaheen Basmati (S. Basmati. Proline concentration increases while chlorophyll “a” chlorophyll “b” and carotene level decreases with increasing salinity. On other hand zinc concentration increases while cadmium and lead concentration decrease in the crop under saline conditions. Intensity of protein polypeptides bands decreases gradually with increasing salinity level but plants from the seeds soaked with smoke solution alleviate the drastic affect of salinity, and intensity of bands is quite good by comparing with non primed seeds. It is concluded that seed priming with plant derived smoke solution show beneficial effect on crop to protect them from salinity.

  4. Proline Accumulation, Photosynthetic Abilities and Growth Characters of Sugarcane (Saccharum officinarum L.) Plantlets in Response to Iso-Osmotic Salt and Water-Deficit Stress

    Suriyan Cha-um; Chalennpol Kirdmanee


    The aim of this study was to investigate the biochemical, physiological and morphological responses of sugarcane to iso-osmotic salt and water-deficit stress. Disease-free sugarcane plantlets derived from meristem cuttings were photo-autotrophically grown in MS media and subsequently exposed to-0.23 (control), -0.67 or -1.20 Mpa iso-osmotic NaCI (salt stress) or mannitol (water-deficit stress). Chlorophyll a (Chl a), chlorophyll b (Chl b), total carotenoids (Cx+c), maximum quantum yield of PSⅡ(Fv/Fm), photon yield of PSⅡ(φPSⅡ), stomatal conductance(Gs)and transpiration rate(E)in the stressed plantlets were significantly reduced when compared to those of plantlets of the control group (without mannitol or NaCI), leading to net-photosynthetic rate (Pn) and growth reduction with positive correlation. In addition, physiological changes and growth parameters of plantlets in the salt stress conditions were more sharply reduced than those in water-deficit stress conditions. On the other hand, the proline content and non-photochemical quenching (NPQ) in the leaves of stressed plantlets increased significantly, especially in response to iso-osmotic salt stress. The chlorophyll pigments in iso-osmotic stressed leaves were significantly degraded (r2=0.93), related to low water oxidation (r2=0.87), low net-photosynthetic rate (r2=0.81), and growth reduction (r2=0.97). The multivariate biochemical, physiological and growth parameters in the present study should be further used to develop salt, or drought, tolerance indices in sugarcane breeding programs.

  5. The effect of phosphours and water deficit on phosphatase activity and proline accumulation in seedling cotyledons and roots of oilseed rape as compared to that of excised cotyledons and roots

    Stanisław Flasiński


    Full Text Available Oilseed rape seedlings and excised cotyledons and roots were exposed to phosphorus and osmotic stress (-1 MPa: NaCl or PEG. The stress factors limited the growth of the seedlings and inhibited the growth of the excised roots and cotyledons. The phosphorus content in the cotyledons and roots depended on its level in the media and on the stress factors used. Phosphorus deficiency differentiated total phosphatase activity in seedling cotyledons and increased the activity in the excised cotyledons. In the excised and seedling roots, the lack of phosphorus, its deficiency and stress imposed by the addition of NaCl, caused an increase in total and specific phosphatase activity. Osmotic stress caused proline accumulation in both the seedling and excised cotyledons, but the effect of phosphorus stress was much smaller. Proline increase in seedling roots followed only after the osmotic stress caused by NaCl. The proline content in the excised roots was low. Stress factors lowered the protein content in the seedling and excised cotyledons. In both seedling and excised roots, the lowering of the protein content occurred mainly in response to osmotic stress. Kinetin modified metabolic responses in seedling cotyledons and roots. The use of in vitro culture allowed the elucidation of the tested responses. In excised cotyledons grown under stress conditions, kinetin increased the phosphorus and protein contents and lowered proline accumulation, suggesting that kinetin had the ability to overcome phosphorus and osmotic stress. The excised oilseed rape roots responded only slightly to kinetin (Drozdowska and Rogozińska 1984 and its effect was much smaller.




    Amino acid transport in right-side-out membrane vesicles of Acinetobacter johnsonii 210A was studied. L-Alanine, L-lysine, and L-proline were actively transported when a proton motive force of -76 mV tvas generated by the oxidation of glucose via the membrane-bound glucose dehydrogenase. Kinetic ana

  7. Effect of Some Stresses on Free Proline Content During Pigeonpea (Cajanas cajan Seed Germination

    Bhamburdekar S.B.


    Full Text Available Plants accumulate proline under a variety of stress conditions thereby preventing stress-caused damages. The proline accumulation in germinating seeds of pigeonpea in response to NaCl, Boron and Aluminium treatments were studied. Results showed increased accumulation of proline in salinity stressed germinating seeds. As compare to 100 ppm boron, 10 and 50 ppm of boron treatments promoted proline levels at different stages of seed germination. The content of proline was found to be induced by all aluminium concentrations particularly at 48, 72 and 120 hrs. of seed germination. Whereas reverse pattern of proline accumulation was observed at 96 hrs stage under all studied stresses.

  8. Proline: Mother Nature's cryoprotectant applied to protein crystallography.

    Pemberton, Travis A; Still, Brady R; Christensen, Emily M; Singh, Harkewal; Srivastava, Dhiraj; Tanner, John J


    L-Proline is one of Mother Nature's cryoprotectants. Plants and yeast accumulate proline under freeze-induced stress and the use of proline in the cryopreservation of biological samples is well established. Here, it is shown that L-proline is also a useful cryoprotectant for protein crystallography. Proline was used to prepare crystals of lysozyme, xylose isomerase, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase for low-temperature data collection. The crystallization solutions in these test cases included the commonly used precipitants ammonium sulfate, sodium chloride and polyethylene glycol and spanned the pH range 4.6-8.5. Thus, proline is compatible with typical protein-crystallization formulations. The proline concentration needed for cryoprotection of these crystals is in the range 2.0-3.0 M. Complete data sets were collected from the proline-protected crystals. Proline performed as well as traditional cryoprotectants based on the diffraction resolution and data-quality statistics. The structures were refined to assess the binding of proline to these proteins. As observed with traditional cryoprotectants such as glycerol and ethylene glycol, the electron-density maps clearly showed the presence of proline molecules bound to the protein. In two cases, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase, proline binds in the active site. It is concluded that L-proline is an effective cryoprotectant for protein crystallography.

  9. Salinity triggers proline synthesis in peanut leaves

    Md. Ashrafuzzaman


    To investigate the magnitude of proline accumulation under different salinity levels, a salinity-imposed experiment was conducted. The peanut seedlings were cultured hydroponically for 7 days before imposing salinity and then grown further for another 7 days under salinity-loaded condition. Results showed that shoot dry matter, relative water content, chlorophyll and K+ decreased significantly with increasing salinity. In contrast, Na+, hydrogen peroxide and proline increased with increasing...

  10. Proline over-accumulation alleviates salt stress and protects photosynthetic and antioxidant enzyme activities in transgenic sorghum [Sorghum bicolor (L.) Moench].

    Surender Reddy, P; Jogeswar, Gadi; Rasineni, Girish K; Maheswari, M; Reddy, Attipalli R; Varshney, Rajeev K; Kavi Kishor, P B


    Shoot-tip derived callus cultures of Sorghum bicolor were transformed by Agrobacterium tumefaciens as well as by bombardment methods with the mutated pyrroline-5-carboxylate synthetase (P5CSF129A) gene encoding the key enzyme for proline biosynthesis from glutamate. The transgenics were selfed for three generations and T4 plants were examined for 100 mM NaCl stress tolerance in pot conditions. The effect of salt stress on chlorophyll and carotenoid contents, photosynthetic rate, stomatal conductance, internal carbon dioxide concentration, transpiration rates, intrinsic transpiration and water use efficiencies, proline content, MDA levels, and antioxidant enzyme activities were evaluated in 40-day-old transgenic lines and the results were compared with untransformed control plants. The results show that chlorophyll content declines by 65% in untransformed controls compared to 30-38% loss (significant at P levels. Photosynthetic rate (PSII activity) was reduced in untransformed controls almost completely, while it declined by 62-88% in different transgenic lines. Salinity induced ca 100% stomatal closure in untransformed plants, while stomatal conductance was decreased only by 64-81% in transgenics after 4 days. The intercellular CO2 decreased by ca 30% in individual transgenic lines. Malondialdehyde (MDA) content was lower in transgenics compared to untransformed controls. The activities of superoxide dismutase (SOD; EC, catalase (CAT; EC and glutathione reductase (GR; EC1.8.1.7) were quantified in leaves exposed to 100 mM NaCl stress and found higher in transgenics. The results suggest that transgenic lines were able to cope better with salt stress than untransformed controls by protecting photosynthetic and antioxidant enzyme activities. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  11. The effect of salinity on chlorophyll, proline and mineral nutrients in common weeds of coastal rice fields in Malaysia.

    Hakim, M A; Juraimi, Abdul Shukor; Hanafi, M M; Ismail, Mohd Razi; Rafii, M Y; Aslani, Fazad; Selamat, A


    Six weed species (Leptochola chinensis, Echinochloa crus-galli, Echinochloa colona, Jussiaea linifolia, Oryza sativa (weedy rice) and Cyperus iria) were tested for their salt tolerant traits in terms of chlorophyll, proline and mineral nutrients accumulation against different salinity levels (0, 4, 8, 16, 24, 32, and 40 dS m(-1)). Chlorophyll a, b and total chlorophyll content, proline and mineral nutrients accumulation were determined. Salt stress showed prominent effect on all the parameters investigated and there were significant variations between the all weed species. Chlorophyll content, K+, Ca(2+) and Mg(2+) ions in both shoots and roots significantly decreased; while proline and Na+ accumulation significantly increased with increasing salinity up to 40 dS m(-1). In terms of overall performance, Cyperus iria and E. crus-galliwere relatively more tolerant; E. colona and J. linifolia were tolerant; L. chinensis and O. sativa L were salt sensitive, respectively.

  12. Penultimate proline in neuropeptides.

    Glover, Matthew S; Bellinger, Earl P; Radivojac, Predrag; Clemmer, David E


    A recent ion mobility spectrometry-mass spectrometry (IMS-MS) study revealed that tryptic peptide ions containing a proline residue at the second position from the N-terminus (i.e., penultimate proline) frequently adopt multiple conformations, owing to the cis-trans isomerization of Xaa(1)-Pro(2) peptide bonds [J. Am. Soc. Mass Spectrom. 2015, 26, 444]. Here, we present a statistical analysis of a neuropeptide database that illustrates penultimate proline residues are frequently found in neuropeptides. In order to probe the effect of penultimate proline on neuropeptide conformations, IMS-MS experiments were performed on two model peptides in which penultimate proline residues were known to be important for biological activity: the N-terminal region of human neuropeptide Y (NPY1-9, Tyr(1)-Pro(2)-Ser(3)-Lys(4)-Pro(5)-Asp(6)-Asn(7)-Pro(8)-Gly(9)-NH2) and a tachykinin-related peptide (CabTRP Ia, Ala(1)-Pro(2)-Ser(3)-Gly(4)-Phe(5)-Leu(6)-Gly(7)-Met(8)-Arg(9)-NH2). From these studies, it appears that penultimate prolines allow neuropeptides to populate multiple conformations arising from the cis-trans isomerization of Xaa(1)-Pro(2) peptide bonds. Although it is commonly proposed that the role of penultimate proline residues is to protect peptides from enzymatic degradation, the present results indicate that penultimate proline residues also are an important means of increasing the conformational heterogeneity of neuropeptides.

  13. Effect of salt stress on growth, inorganic ion and proline ...



    Jan 11, 2010 ... Key word: Aromatic rice, osmoprotectant, callus culture, salt stress. ... adversely affect the overall metabolic activities and cause plant demise (Roychoudury ..... Proline accumulation in response to environmental stresses has ...

  14. Influence of irradiation parameters on damage accumulation in metals and alloys

    Singh, B.N.; Zinkle, S.J.


    It is well known that a fraction of defects produced during irradiation accumulate in crystalline solids in the form of clusters of self-interstitial atoms (SIAs) and vacancies, loops, tetrahedra, dislocation segments and cavities. The irradiation parameters such as recoil energy, damage rate, he...

  15. Genetically modified cyanobacterium Nostoc muscorum overproducing proline in response to salinity and osmotic stresses

    Santosh Bhargava


    In the parent Nostoc muscorum an active proline oxidase enzyme is required to assimilate exogenous proline as a fixed nitrogen source. Cyanobacterial mutants, resistant to growth inhibitory action of proline analogue L-azetidine-2-carboxylate (Ac-R), were deficient in proline oxidase activity, and were over-accumulators of proline. Proline over-accumulation, resulting either from mutational acquisition of the Ac-R phenotype, or from salinity-induced uptake of exogenous proline, confirmed enhanced salinity/osmotic tolerance in the mutant strain. The nitrogenase activity and photosynthetic O2 evolution of the parent were sensitive to both salinity as well as osmotic stresses than of Ac-R mutant strain. In addition, the mutation to Ac-resistant phenotype showed no alteration in salinity inducible potassium transport system in the cyanobacterium

  16. Solid state parameters, structure elucidation, High Resolution X-Ray Diffraction (HRXRD), phase matching, thermal and impedance analysis on L-Proline trichloroacetate (L-PTCA) NLO single crystals.

    Kalaiselvi, P; Raj, S Alfred Cecil; Jagannathan, K; Vijayan, N; Bhagavannarayana, G; Kalainathan, S


    Nonlinear optical single crystal of L-Proline trichloroacetate (L-PTCA) was successfully grown by Slow Evaporation Solution Technique (SEST). The grown crystals were subjected to single crystal X-ray diffraction analysis to confirm the structure. From the single crystal XRD data, solid state parameters were determined for the grown crystal. The crystalline perfection has been evaluated using high resolution X-ray diffractometer. The frequencies of various functional groups were identified from FTIR spectral analysis. The percentage of transmittance was obtained from UV Visible spectral analysis. TGA-DSC measurements indicate the thermal stability of the crystal. The dielectric constant, dielectric loss and ac conductivity were measured by the impedance analyzer. The DC conductivity was calculated by the cole-cole plot method.

  17. Proline improves copper tolerance in chickpea (Cicer arietinum).

    Singh, Vijeta; Bhatt, Indu; Aggarwal, Anjali; Tripathi, Bhumi Nath; Munjal, Ashok Kumar; Sharma, Vinay


    The present study suggests the involvement of proline in copper tolerance of four genotypes of Cicer arietinum (chickpea). Based on the data of tolerance index and lipid peroxidation, the order for copper tolerance was as follows: RSG 888 > CSG 144 > CSG 104 > RSG 44 in the selected genotypes. The basis of differential copper tolerance in chickpea genotypes was characterized by analyzing, antioxidant enzymes (superoxide dismutase, ascorbated peroxidase and catalase), phytochelatins, copper uptake, and proline accumulation. Chickpea genotypes showed stimulated superoxide dismutase activity at all tested concentrations of copper, but H(2)O(2) decomposing enzymes especially; ascorbate peroxidase did not increase with 25 and 50 microM copper treatments. Catalase activity, however, increased at lower copper concentrations but failed to stimulate at 50 microM copper. Such divergence in responses of these enzymes minimizes their importance in protecting chickpea against copper stress. The sensitive genotypes showed greater enhancement of phytochelatins than that of tolerant genotypes. Hence, the possibility of phytochelatins in improving copper tolerance in the test plant is also excluded. Interestingly, the order of proline accumulation in the chickpea genotypes (RSG 888 > CSG 144 > CSG 104 > RSG 44) was exactly similar to the order of copper tolerance. Based on hyperaccumulation of proline in tolerant genotype (RSG 44) and the reduction and improvement of lipid peroxidation and tolerance index, respectively, by proline pretreatment, we conclude that hyperaccumulation of proline improves the copper tolerance in chickpea.

  18. Continuous proline catalysis via leaching of solid proline

    Suzanne M. Opalka


    Full Text Available Herein, we demonstrate that a homogeneous catalyst can be prepared continuously via reaction with a packed-bed of a catalyst precursor. Specifically, we perform continuous proline catalyzed α-aminoxylations using a packed-bed of L-proline. The system relies on a multistep sequence in which an aldehyde and thiourea additive are passed through a column of solid proline, presumably forming a soluble oxazolidinone intermediate. This transports a catalytic amount of proline from the packed-bed into the reactor coil for subsequent combination with a solution of nitrosobenzene, affording the desired optically active α-aminooxy alcohol after reduction. To our knowledge, this is the first example in which a homogeneous catalyst is produced continuously using a packed-bed. We predict that the method will not only be useful for other L-proline catalyzed reactions, but we also foresee that it could be used to produce other catalytic species in flow.

  19. Application of cinchona-sulfonate-based chiral zwitterionic ion exchangers for the separation of proline-containing dipeptide rotamers and determination of on-column isomerization parameters from dynamic elution profiles

    Wernisch, Stefanie, E-mail: [Institute for Analytical Chemistry, University of Vienna, Währinger Straße 38, A-1090 Vienna (Austria); Trapp, Oliver, E-mail: [Organisch-Chemisches Institut, Ruprecht-Karls-Universität Heidelberg, Im Neuenheimer Feld 270, D-69120 Heidelberg (Germany); Lindner, Wolfgang, E-mail: [Institute for Analytical Chemistry, University of Vienna, Währinger Straße 38, A-1090 Vienna (Austria)


    Graphical abstract: -- Highlights: •X-Pro peptides produce interconverting elution profiles on zwitterionic stationary phases. •The cis–trans isomerization rate in solution is lower than in the chromatographic system. •Cinchona-based zwitterionic selectors apparently lower the isomerization barrier. •Rotamer separation is feasible <0 °C, enantiomer separation above room temperature. -- Abstract: The interconversion of cis and trans isomers of dipeptides containing C-terminal proline was studied by dynamic chromatography on zwitterionic chiral stationary phases at temperatures ranging from −15 °C to +45 °C The cis–trans isomers could be separated below 0 °C and above 0–10 °C plateau formation and peak coalescence phenomena occurred, which is characteristic for a dynamic process at the time-scale of partitioning. At and above room temperature, full coalescence was observed, which allowed separations of enantiomers without interference from interconversion effects. Analysis of the dynamic elution profiles of the interconverting peptides allowed the determination of isomerization rate constants and thermodynamic activation parameters (isomerization enthalpy, entropy and activation energy). In accordance with established results, isomerization rates and thermodynamic parameters were found to depend on the nature of the N-terminal amino acid. Isomerization barriers were only slightly lower than values determined with other methods but significant differences in the relative contributions of the activation enthalpy and entropy as well as isomerization rates pointed toward selector-moderated isomerization dynamics.

  20. Application of cinchona-sulfonate-based chiral zwitterionic ion exchangers for the separation of proline-containing dipeptide rotamers and determination of on-column isomerization parameters from dynamic elution profiles.

    Wernisch, Stefanie; Trapp, Oliver; Lindner, Wolfgang


    The interconversion of cis and trans isomers of dipeptides containing C-terminal proline was studied by dynamic chromatography on zwitterionic chiral stationary phases at temperatures ranging from -15°C to +45°C The cis-trans isomers could be separated below 0°C and above 0-10°C plateau formation and peak coalescence phenomena occurred, which is characteristic for a dynamic process at the time-scale of partitioning. At and above room temperature, full coalescence was observed, which allowed separations of enantiomers without interference from interconversion effects. Analysis of the dynamic elution profiles of the interconverting peptides allowed the determination of isomerization rate constants and thermodynamic activation parameters (isomerization enthalpy, entropy and activation energy). In accordance with established results, isomerization rates and thermodynamic parameters were found to depend on the nature of the N-terminal amino acid. Isomerization barriers were only slightly lower than values determined with other methods but significant differences in the relative contributions of the activation enthalpy and entropy as well as isomerization rates pointed toward selector-moderated isomerization dynamics.

  1. Atmospheric dust accumulation on native and non-native species: effects on gas exchange parameters.

    González, Juan A; Prado, Fernando E; Piacentini, Ruben D


    Plants are continuously exposed to atmospheric particulate matter (dust), and their leaves are the main receptors of deposited dust. The objective of this study was to assess the effects of dust deposition on leaf gas exchange parameters of 17 native and non-native tree and shrub species growing in Gran San Miguel de Tucumán in northwestern Argentina. Maximum assimilation rate (), stomatal conductance (), transpiration rate (), internal CO concentration (), and instantaneous water-use efficiency (WUE) were measured in cleaned leaves (CL) and dusted leaves (DL) of different species on November 2010, July 2011, and September 2011. In almost all studied species, gas exchange parameters were significantly affected by dust deposition. Values for , , and of DL were significantly reduced in 11, 12, and 14 species compared with CL. Morphological leaf traits seem to be related to reduction. Indeed, L. and (Mart. ex DC.) Standl. species with pubescent leaves and thick ribs showed the highest reduction percentages. Contrarily, and WUE were increased in DL but were less responsive to dust deposition than other parameters. Increases of and WUE were significant in 5 and 11 species, respectively. Correlation analyses between /, /, and / pairs showed significant positive linear correlations in CL and DL of many studied species, including small and tall plants. These results suggest that leaf stomatal factors and shade-induced effect by accumulated dust are primarily responsible for the observed reductions in photosynthesis rate of DL.

  2. Astaxanthin Accumulation in the Green Alga Haematococcus pluvialis: Effects of Cultivation Parameters

    Ping He; James Duncan; James Barber


    The green alga, Haematococcus pluvialis Flotow is used as a source of the ketocarotenoid astaxanthin for application in fish aquaculture, pharmaceutical and cosmetic industries. Cells of the green alga were induced by the application of different light and starvation conditions to evaluate the effect in astaxanthin accumulate. The condiphosphate starvation. The results show that stresses applied in culture, which interfere with cell division, trigger the accumulation of astaxanthin. Notably, sulfur starvation results in a massive accumulation of this commercially important carotenoid.

  3. Phytoremediation of cadmium improved with the high production of endogenous phenolics and free proline contents in Parthenium hysterophorus plant treated exogenously with plant growth regulator and chelating agent.

    Ali, Nasir; Hadi, Fazal


    Pot experiments were conducted to evaluate the effects of gibberellic acid (GA3) and ethylenediaminetetraacetic acid (EDTA) on growth parameters, cadmium (Cd) phytoextraction, total phenolics, free proline and chlorophyll content of Parthenium hysterophorus plant grown in Cd-contaminated (100 mg/kg) soil. GA3 was applied as foliar spray (10(-2), 10(-4) and 10(-6) M) while EDTA (40 mg/kg soil) was added to soil as single and in split doses. Results showed decrease in growth parameters due to Cd stress but P. hysterophorus plant demonstrated Cd hyperaccumulator potential based on bioconcentration factor (BCF). Lower concentration of GA3 (10(-6) M) showed highest significant increase in the growth parameters while Cd concentration, accumulation (1.97 ± 0.11 mg/DBM) and bioconcentration (9.75 ± 0.34) was significantly higher in the treatment T11 (GA3 10(-2) + split doses of EDTA). Cadmium significantly increased the root free proline while total phenolic concentration was significantly high in all parts of the plant. Chlorophyll contents were significantly reduced by Cd. GA3 showed significant increase in phenolic and chlorophyll contents in plant. Cadmium accumulation in plant tissues showed positive correlation with free proline (R (2) = 0.527, R (2) = 0.630) and total phenolics (R (2) = 0.554, R (2) = 0.723) in roots and leaves, respectively. Cd contents negatively correlated with biomass, chlorophyll and total water contents. Proline and phenolic contents showed positive correlation with dry biomass of plant. These findings suggest further investigation to study the role of endogenous phenolics and proline in heavy metal phytoremediation.

  4. Effects of Nitrogen and Shading on Root Morphologies, Nutrient Accumulation, and Photosynthetic Parameters in Different Rice Genotypes.

    Pan, Shenggang; Liu, Haidong; Mo, Zhaowen; Patterson, Bob; Duan, Meiyang; Tian, Hua; Hu, Shuijing; Tang, Xiangru


    Nitrogen availability and illumination intensity are two key factors which affect rice growth. However, their influences on total nitrogen accumulation, photosynthetic rate, root morphologies, and yields are not fully understood. We conducted two field experiments to (1) evaluate the effects of shading under different N treatments on photosynthetic parameters, root morphologies, total nutrient accumulation, and grain yields of rice; and (2) elucidate the relationship between total nutrient accumulation and root morphologies under different shading conditions and nitrogen treatments. Three nitrogen rates, three shading treatments, and three different rice cultivars were used in two field experiments. Double shading during the grain-filling stage decreased total nutrient accumulation, altered root morphological characteristics, and decreased yields in rice. There were also significant interaction effects between nitrogen and shading on photosynthetic rate, transpiration rate, and total root length, root superficial area, and root volume. Significant interactions were found among cultivars and shading for photosynthetic rate and transpiration rate. Correlation analysis revealed that total nitrogen accumulation (TNA) and potassium accumulation (TKA) were significantly positively correlated with total root length, root superficial area, and root volume. N application could alleviate the detrimental effects of shading on total nutrient accumulation and grain yield in rice.

  5. Effects of the Lycopersicon chmielewskii sucrose accumulator gene (sucr) on fruit yield and quality parameters following introgression into tomato.

    Chetelat, R T; Deverna, J W; Bennett, A B


    A gene controlling fruit sucrose accumulation, sucr, was introgressed from the wild tomato species Lycopersicon chmielewskii into the genetic background of a hexose-accumulating cultivated tomato, L. esculentum. During introgression, the size of the L. chmielewskii chromosomal segment containing sucr was reduced by selection for recombination between RFLP markers for the sucr gene and flanking loci. The effects of sucr on soluble solids content, fruit size, yield and other fruit parameters were studied in the genetic background of the processing tomato cultivar 'Huntl00'. In a segregating BC5F2 generation, the smallest introgression containing sucr-associated markers was necessary and sufficient to confer high-level sucrose accumulation, the effects of which were completely recessive. Fruit of sucr/sucr genotypes were smaller than those of +/sucr or +/+ genotypes at all stages of development. The timing of sugar accumulation and total sugar concentration were unaffected by sugar composition. No differences in total fruit biomass (fresh weight of red and green fruit) at harvest were observed between the genotypes, and sucrose accumulators produced greater numbers of fruit than hexose accumulators in one family. However, the proportion of ripe fruit at harvest, and hence yield of ripe fruit, as well as average ripe fruit weight and seed set were reduced in sucr/sucr genotypes. Sucrose accumulation was also associated with increased soluble solids content, consistency, serum viscosity, predicted paste yield and acidity, and decreased color rating. In the first backcross to L. chmielewskii, hexose accumulators (+/sucr) had larger fruit than sucrose accumulators (sucr/sucr), while no difference in soluble solids was detected.

  6. Requirement of proline synthesis during Arabidopsis reproductive development

    Funck Dietmar


    Full Text Available Abstract Background Gamete and embryo development are crucial for successful reproduction and seed set in plants, which is often the determining factor for crop yield. Proline accumulation was largely viewed as a specific reaction to overcome stress conditions, while recent studies suggested important functions of proline metabolism also in reproductive development. Both the level of free proline and proline metabolism were proposed to influence the transition to flowering, as well as pollen and embryo development. Results In this study, we performed a detailed analysis of the contribution of individual proline biosynthetic enzymes to vegetative development and reproductive success in Arabidopsis. In contrast to previous reports, we found that pyrroline-5-carboxylate (P5C synthetase 2 (P5CS2 is not essential for sexual reproduction although p5cs2 mutant plants were retarded in vegetative development and displayed reduced fertility under long-day conditions. Single mutant plants devoid of P5CS1 did not show any developmental defects. Simultaneous absence of both P5CS isoforms resulted in pollen sterility, while fertile egg cells could still be produced. Expression of P5C reductase (P5CR was indispensable for embryo development but surprisingly not needed for pollen or egg cell fertility. The latter observation could be explained by an extreme stability of P5CR activity, which had a half-life time of greater than 3 weeks in vitro. Expression of P5CR-GFP under the control of the endogenous P5CR promoter was able to restore growth of homozygous p5cr mutant embryos. The analysis of P5CR-GFP-fluorescence in planta supported an exclusively cytoplasmatic localisation of P5CR. Conclusions Our results demonstrate that potential alternative pathways for proline synthesis or inter-generation transfer of proline are not sufficient to overcome a defect in proline biosynthesis from glutamate during pollen development. Proline biosynthesis through P5CS2 and P5

  7. Involvement of Phosphatidylinositol 3-kinase in the regulation of proline catabolism in Arabidopsis thaliana

    Anne-Sophie eLeprince


    Full Text Available Plant adaptation to abiotic stresses such as drought and salinity involves complex regulatory processes. Deciphering the signalling components that are involved in stress signal transduction and cellular responses is of importance to understand how plants cope with salt stress. Accumulation of osmolytes such as proline is considered to participate in the osmotic adjustment of plant cells to salinity. Proline accumulation results from a tight regulation between its biosynthesis and catabolism. Lipid signal components such as phospholipases C and D have previously been shown to be involved in the regulation of proline metabolism in Arabidopsis thaliana. In this study, we demonstrate that proline metabolism is also regulated by class-III Phosphatidylinositol 3-kinase (PI3K, VPS34, which catalyses the formation of phosphatidylinositol 3-phosphate (PI3P from phosphatidylinositol. Using pharmacological and biochemical approaches, we show that the PI3K inhibitor, LY294002, affects PI3P levels in vivo and that it triggers a decrease in proline accumulation in response to salt treatment of A. thaliana seedlings. The lower proline accumulation is correlated with a lower transcript level of Pyrroline-5-carboxylate synthetase 1 biosynthetic enzyme and higher transcript and protein levels of Proline dehydrogenase 1 (ProDH1, a key-enzyme in proline catabolism. We also found that the ProDH1 expression is induced in a pi3k-hemizygous mutant, further demonstrating that PI3K is involved in the regulation of proline catabolism through transcriptional regulation of ProDH1. A broader metabolomic analysis indicates that LY294002 also reduced other metabolites, such as hydrophobic and aromatic amino acids and sugars like raffinose.

  8. Role of proline in cell wall synthesis and plant development and its implications in plant ontogeny



    Full Text Available Proline is a proteogenic amino acid and accumulates both under stress and non-stress conditions as a beneficial solute in plants. Recent discoveries point out that proline plays an important role in plant growth and differentiation across life cycle. It is a key determinant of many cell wall proteins that plays important roles in plant development. The role of extensins (EXTs, arabinogalactan proteins (AGPs and hydroxyproline- and proline-rich proteins (H/PRPs as important components of cell wall proteins that play pivotal roles in cell wall signal transduction cascades, plant development and stress tolerance is discussed in this review. Molecular insights are also provided here into the plausible roles of proline transporters modulating key events in plant development. In addition, the roles of proline during seed developmental transitions including storage protein synthesis are discussed.

  9. Response of Two Tomato Cultivars to Field-applied Proline and Salt Stress

    Kahlaoui B.


    Full Text Available An experiment was carried out using saline water (6.57 dS.m-1 and subsurface drip irrigation (SDI on two tomato cultivars (Solanum lycopersicum, cv. Rio Grande and Heinz-2274 in a silty clay soil. The former is a salinity tolerant and the latter a sensitive cultivar. Exogenous application of proline was done by foliar spray at two concentrations: 10 and 20 mg.L-1, with a control (saline water without proline, during the flowering stage. As a result of the proline applied, significant effects were observed on both cultivars of tomato, particularly with low concentration of proline (10 mg.L-1. It led to increase of leaf area, growth length and fruit yield. Regarding mineral nutrition, Ca2+ was higher in different organs while low accumulation of Na+ occurred. However, Cl- was very low significantly in all tissues of plants of Rio Grande at the higher concentration of proline applied.

  10. The influence of bioavailable heavy metals and microbial parameters of soil on the metal accumulation in rice grain.

    Xiao, Ling; Guan, Dongsheng; Peart, M R; Chen, Yujuan; Li, Qiqi; Dai, Jun


    A field-based study was undertaken to analyze the effects of soil bioavailable heavy metals determined by a sequential extraction procedure, and soil microbial parameters on the heavy metal accumulation in rice grain. The results showed that Cd, Cr, Cu, Ni, Pb and Zn concentrations in rice grain decreases by 65.9%, 78.9%, 32.6%, 80.5%, 61.0% and 15.7%, respectively in the sites 3 (far-away), compared with those in sites 1 (close-to). Redundancy analysis (RDA) indicated that soil catalase activity, the MBC/MBN ratio, along with bioavailable Cd, Cr and Ni could explain 68.9% of the total eigenvalue, indicating that these parameters have a great impact on the heavy metal accumulation in rice grain. The soil bioavailable heavy metals have a dominant impact on their accumulation in rice grain, with a variance contribution of 60.1%, while the MBC/MBN has a regulatory effect, with a variance contribution of 4.1%. Stepwise regression analysis showed that the MBC/MBN, urease and catalase activities are the key microbial parameters that affect the heavy metal accumulation in rice by influencing the soil bioavailable heavy metals or the translocation of heavy metals in rice. RDA showed an interactive effect between Cu, Pb and Zn in rice grain and the soil bioavailable Cd, Cr and Ni. The heavy metals in rice grain, with the exception of Pb, could be predicted by their respective soil bioavailable heavy metals. The results suggested that Pb accumulation in rice grain was mainly influenced by the multi-metal interactive effects, and less affected by soil bioavailable Pb. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Simple DVH parameter addition as compared to deformable registration for bladder dose accumulation in cervix cancer brachytherapy

    Andersen, Else Stougård; Noe, Karsten Østergaaard; Sørensen, Thomas Sangild


    called "the worst case assumption") in fractionated BT. Materials and methods: Forty-seven patients treated for locally advanced cervical cancer were included. All patients received EBRT combined with two individually planned 3D image-guided adaptive BT fractions. D2 and D0.1 were estimated by DVH...... parameter addition and compared to dose accumulations based on an in-house developed biomechanical deformable image registration (DIR) algorithm. Results: DIR-based DVH analysis was possible in 42/47 patients. DVH parameter addition resulted in mean dose deviations relative to DIR of 0.4 ± 0.3 Gy (1.5 ± 1...

  12. Functional specialization in proline biosynthesis of melanoma.

    Jessica De Ingeniis

    Full Text Available Proline metabolism is linked to hyperprolinemia, schizophrenia, cutis laxa, and cancer. In the latter case, tumor cells tend to rely on proline biosynthesis rather than salvage. Proline is synthesized from either glutamate or ornithine; both are converted to pyrroline-5-carboxylate (P5C, and then to proline via pyrroline-5-carboxylate reductases (PYCRs. Here, the role of three isozymic versions of PYCR was addressed in human melanoma cells by tracking the fate of (13C-labeled precursors. Based on these studies we conclude that PYCR1 and PYCR2, which are localized in the mitochondria, are primarily involved in conversion of glutamate to proline. PYCRL, localized in the cytosol, is exclusively linked to the conversion of ornithine to proline. This analysis provides the first clarification of the role of PYCRs to proline biosynthesis.

  13. Functional Specialization in Proline Biosynthesis of Melanoma

    Richardson, Adam D.; Scott, David A.; Aza-Blanc, Pedro; De, Surya K.; Kazanov, Marat; Pellecchia, Maurizio; Ronai, Ze'ev; Osterman, Andrei L.; Smith, Jeffrey W.


    Proline metabolism is linked to hyperprolinemia, schizophrenia, cutis laxa, and cancer. In the latter case, tumor cells tend to rely on proline biosynthesis rather than salvage. Proline is synthesized from either glutamate or ornithine; both are converted to pyrroline-5-carboxylate (P5C), and then to proline via pyrroline-5-carboxylate reductases (PYCRs). Here, the role of three isozymic versions of PYCR was addressed in human melanoma cells by tracking the fate of 13C-labeled precursors. Based on these studies we conclude that PYCR1 and PYCR2, which are localized in the mitochondria, are primarily involved in conversion of glutamate to proline. PYCRL, localized in the cytosol, is exclusively linked to the conversion of ornithine to proline. This analysis provides the first clarification of the role of PYCRs to proline biosynthesis. PMID:23024808

  14. Enhancement of anthraquinone production in Morinda citrifolia cell suspension cultures after stimulation of the proline cycle with two proline analogs.

    Quevedo, Carla V; Perassolo, María; Giulietti, Ana M; Rodríguez Talou, Julián


    Synthesis of anthraquinones (AQs) involves the shikimate and 2-C-methyl-D-erythritol 4-phosphate pathways. The proline cycle is linked to the pentose phosphate pathway (PPP) to generate NADPH needed in the first steps of this pathway. The effect of two proline analogs, azetidine-2-carboxylic acid (A2C) and thiazolidine-4-carboxylic acid (T4C), were evaluated in Morinda citrifolia suspension cultures. Both analogs gave higher proline accumulation after 6 and 10 days (68 and 179% after 6 days with A2C at 25 and 50 μM, respectively, and 111% with T4C added at 100 μM). Induction of the proline cycle increased the AQ content after 6 days (~40% for 50 μM A2C and 100 μM T4C). Whereas A2C (50 μM) increased only AQ production, T4C also enhanced total phenolics. However, no induction of the PPP was observed with any of the treatments. This pathway therefore does not limit the supply of carbon skeletons to secondary metabolic pathways.

  15. Proline is a protein solubilizing solute.

    Samuel, D; Kumar, T K; Jayaraman, G; Yang, P W; Yu, C


    The effect of proline on the prevention of trichloroacetic acid (TCA)-induced protein precipitation is studied. It is found that proline at high concentrations (> 4.0 M) completely prevents TCA-induced precipitation of hen egg white lysozyme. Other osmolytes such as ethylene glycol, glycerol and sucrose fail to prevent the TCA-induced precipitation of lysozyme. Viscosity and 1-anilino-8-naphthalene sulphonic acid binding experiments suggest that proline at high concentration forms an ordered supramolecular assembly. Proline is shown to increase the solubility of protein due to formation of such higher order assemblies. A model of the supra-molecular assembly of proline is proposed and a possible in vivo role of the increased levels of proline under water stress is discussed.

  16. Temperatura, umidade relativa e disponibilidade de água no acumulo de prolina em cultivares de feijoeiro Temperature, relative humidity and water availability on proline accumulation of dry beans

    Haiko Enok Sawazaki


    Full Text Available O estudo de plantas de feijão (Phaseolus vulgaris L. submetidas à deficiência hídrica e temperaturas de 35-40º C umidade relativa (UR 40-100%; 5-10ºC, UR 20-30%; 2-5ºC, UR 10-20%, realizado em casa de vegetação e câmara de crescimento, no Instituto Agronômico, em 1982, mostrou haver correlação negativa entre o acúmulo de prolina e o potencial de pressão de água nas folhas (psip. Os cultivares Curitibano e P-511, este último considerado resistente à seca e baixa temperatura, apresentaram o menor acumulo de prolina e o maior psip, confirmando a hipótese de que é possível o reconhecimento de material geneticamente resistente à escassez de água através da seleção de plantas que acumulam menor quantidade de prolina.Four experiments were performed in growth chamber and greenhouse at Campinas, in 1982, to study the relations of dry bean cultivars submmited to period of water and temperatura stress. Adaptation of planta to stress conditions was measured through prolina accumulation and water potential (psip in leaves. In all tested cultivars prolina content showed a straight negative correlation with water potential in leaves. Cultivars P-511 and Curitibano showed lower prolina content and higher water potential than the other under study, this confirming the hypothesis that it is possible to identify materiais genetically resistant to water stress by selection of plants or lines with low prolina content.

  17. Estimation of secondary effect parameters in glycaemic dynamics using accumulating data from a virtual type 1 diabetic patient.

    Mansell, Erin J; Docherty, Paul D; Fisk, Liam M; Chase, J Geoffrey


    Some individuals with type 1 diabetes mellitus find self-managed glycaemic control difficult due to the confounding influence of secondary effects. Stress and sleep deprivation temporarily lower insulin sensitivity (SI), often resulting in hyperglycaemia, while aerobic exercise depletes glucose, leading to hypoglycaemia if treatment is unchanged. This study tests the estimation of these factors and circadian rhythms of SI in noisy data. Sparse, irregular and noisy virtual blood glucose data, mimicking the glycaemic dynamics of an individual with type 1 diabetes, was created via adapted pharmacokinetic-pharmacodynamic models of glucose and insulin that included the impact of the secondary effects. A Gauss-Newton algorithm was used to recover the original model parameters for SI, stress, fatigue and exercise. During longer identification periods, compensation was made for drift in SI. Monte Carlo analyses were undertaken to validate the methods. The coefficient of variation (CV) in all parameters decreased as the data accumulated in proportion to the 1/n rule (R(2) > 99.9%). Relatively small biases from the original parameter values occurred (data to override the effect of measurement error. Compensation for SI drift allows viable observation of secondary effects and SI rhythms over longer time periods. Collectively, these outcomes indicate that quality results for identified parameters could be obtained during in vivo studies.

  18. Preparation of Spirocyclic β-Proline Esters

    Fjelbye, Kasper; Marigo, Mauro; Clausen, Rasmus Prætorius


    A series of novel N-Bn-protected spirocyclic β-proline esters were prepared using [3+2] cycloaddition and subsequently converted into their corresponding aldehydes. In addition, two novel N-Cbz-protected spirocyclic β-proline esters were prepared using intramolecular cyclization starting from...

  19. Trimming proline dehydrogenase : protein and cofactor minimization

    Huijbers, Mieke M.E.


    Proline is one of the proteinogenic amino acids and one of the most abundant amino acids in the cell. Next to serving as one of the non-essential amino acids, proline also has a central role in metabolism. In Chapter 1, the different functions of this imino acid are described, as we

  20. Interaction between cadmium and iron. Accumulation and distribution of metals and changes in growth parameters of Phaseolus vulgaris L. seedlings

    Anna Siedlecka


    Full Text Available The interaction between cadmium, one of the most toxic heavy metals, and iron, an essential plant nutritional element, was investigated in Phaseolus vulgaris L. (cv. Słowianka seedlings. The interaction was externally induced by changing the content of both metals in the nutrient medium. Under iron deficiency conditions (0 and 0.5 of normal dose of this element, the toxic effects of cadmium on plant growth parameters, like fresh and dry weight accumulation, primary leaves area, etc., were generally much more pronounced than under normal iron supply. At normal and excess iron supply (1, 2 and 4 doses cadmium diminished iron accumulation in roots and primary leaves, but on the other hand excess iron decreased cadmium level, preventing plants from extreme toxicity of very high cadmium concentrations in the growth environment. It is to be noted that iron is classified also as a heavy metal, and its excess may become toxic, e.g. decreasing root dry weight or diminishing leaf area, especially at the highest dose. The detoxication role of iron against cadmium, and possibly other toxic metals is, however, limited to concentrations of this element in the nutrient solution which themselves are not toxic for the organism.

  1. Screening of Argentine plant extracts: impact on growth parameters and aflatoxin B1 accumulation by Aspergillus section Flavi.

    Bluma, R; Amaiden, M R; Etcheverry, M


    The effect of essential oils, ethanolic and aqueous extract of 41 vegetable species on Aspergillus section Flavi growth was evaluated. The in vitro screen was a two-stage process. A wide-spectrum initial screen which identified promising antifungal plant extracts was carried out first. After that, identified extracts were studied in more detail by in vitro assays. A total of 96 plant extracts were screened. Essential oils were found to be the most effective extract controlling aflatoxigenic strains. Clove, mountain thyme, poleo and eucalyptus essential oils were selected to study their antifungal effect. Studies on percentage of germination, germ-tube elongation rate, growth rate, and aflatoxin B1 accumulation were carried out. Clove, mountain thyme and poleo essential oils showed the most antifungal effect under all growth parameters analyzed as well as aflatoxin B1 accumulation. Our results suggest that mountain thyme and poleo, which are native vegetal species of Argentina, and clove essential oils could be used alone or in conjunction with other substances to control the presence of aflatoxigenic fungi in stored maize.

  2. Hydrogen sulfide donor sodium hydrosulfide-improved heat tolerance in maize and involvement of proline.

    Li, Zhong-Guang; Ding, Xiao-Jiao; Du, Pei-Fang


    Hydrogen sulfide (H2S) has long been considered as a phytotoxin, but nowadays as a cell signal molecule involved in growth, development, and the acquisition of stress tolerance in higher plants. In the present study, hydrogen sulfide donor, sodium hydrosulfide (NaHS), pretreatment markedly improved germination percentage of seeds and survival percentage of seedlings of maize under heat stress, and alleviated an increase in electrolyte leakage of roots, a decrease in tissue vitality and an accumulation of malondialdehyde (MDA) in coleoptiles of maize seedlings. In addition, pretreatment of NaHS could improve the activity of Δ(1)-pyrroline-5-carboxylate synthetase (P5CS) and lower proline dehydrogenase (ProDH) activity, which in turn induced accumulation of endogenous proline in maize seedlings. Also, application of proline could enhance endogenous proline content, followed by mitigated accumulation of MDA and increased survival percentage of maize seedlings under heat stress. These results suggest that sodium hydrosulfide pretreatment could improve heat tolerance of maize and the acquisition of this heat tolerance may be involved in proline.

  3. Effect of water regime on the growth, flower yield, essential oil and proline contents of Calendula officinalis



    Full Text Available Metwally SA,Khalid KA, Abou-Leila BH. 2013. Effect of water regime on the growth, flower yield, essential oil and proline contents of Calendula officinalis. Nusantara Bioscience 5: 63-67. The effects of water regime on the growth, content of essential oil and proline of Calendula officinalis L. plants were investigated. Water regimes of 75% of field water capacity increased certain growth characters [i.e. plant height (cm, leaf area (cm2, flower diameter (cm and spike stem diameter] and vase life (day. Water regime promoted the accumulation of essential oil content and its main components as well as proline contents.

  4. On the Split Personality of Penultimate Proline

    Glover, Matthew S.; Shi, Liuqing; Fuller, Daniel R.; Arnold, Randy J.; Radivojac, Predrag; Clemmer, David E.


    The influence of the position of the amino acid proline in polypeptide sequences is examined by a combination of ion mobility spectrometry-mass spectrometry (IMS-MS), amino acid substitutions, and molecular modeling. The results suggest that when proline exists as the second residue from the N-terminus (i.e., penultimate proline), two families of conformers are formed. We demonstrate the existence of these families by a study of a series of truncated and mutated peptides derived from the 11-residue peptide Ser1-Pro2-Glu3-Leu4-Pro5-Ser6-Pro7-Gln8-Ala9-Glu10-Lys11. We find that every peptide from this sequence with a penultimate proline residue has multiple conformations. Substitution of Ala for Pro residues indicates that multiple conformers arise from the cis- trans isomerization of Xaa1-Pro2 peptide bonds as Xaa-Ala peptide bonds are unlikely to adopt the cis isomer, and examination of spectra from a library of 58 peptides indicates that ~80% of sequences show this effect. A simple mechanism suggesting that the barrier between the cis- and trans-proline forms is lowered because of low steric impedance is proposed. This observation may have interesting biological implications as well, and we note that a number of biologically active peptides have penultimate proline residues.

  5. Solid-Phase Synthesis of Amine/Carboxyl Substituted Prolines and Proline Homologues: Scope and Limitations.

    Zhou, Ziniu; Scott, William L; O'Donnell, Martin J


    A solid-phase procedure is used to synthesize racemic peptidomimetics based on the fundamental peptide unit. The peptidomimetics are constructed around proline or proline homologues variably substituted at the amine and carbonyl sites. The procedure expands the diversity of substituted peptidomimetic molecules available to the Distributed Drug Discovery (D3) project. Using a BAL-based solid-phase synthetic sequence the proline or proline homologue subunit is both constructed and incorporated into the peptidomimetic by an α-alkylation, hydrolysis and intramolecular cyclization sequence. Further transformations on solid-phase provide access to a variety of piperazine derivatives representing a class of molecules known to exhibit central nervous system activity. The procedure works well with proline cores, but with larger six- and seven-membered ring homologues the nature of the carboxylic acid acylating the cyclic amine can lead to side reactions and result in poor overall yields.

  6. Mycorrhizas alter sucrose and proline metabolism in trifoliate orange exposed to drought stress

    Wu, Hui-Hui; Zou, Ying-Ning; Rahman, Mohammed Mahabubur; Ni, Qiu-Dan; Wu, Qiang-Sheng


    Arbuscular mycorrhizal fungi (AMF) can enhance drought tolerance in plants, whereas little is known regarding AMF contribution to sucrose and proline metabolisms under drought stress (DS). In this study, Funneliformis mosseae and Paraglomus occultum were inoculated into trifoliate orange (Poncirus trifoliata) under well watered and DS. Although the 71-days DS notably (P < 0.05) inhibited mycorrhizal colonization, AMF seedlings showed significantly (P < 0.05) higher plant growth performance and leaf relative water content, regardless of soil water status. AMF inoculation significantly (P < 0.05) increased leaf sucrose, glucose and fructose concentration under DS, accompanied with a significant increase of leaf sucrose phosphate synthase, neutral invertase, and net activity of sucrose-metabolized enzymes and a decrease in leaf acid invertase and sucrose synthase activity. AMF inoculation produced no change in leaf ornithine-δ-aminotransferase activity, but significantly (P < 0.05) increased leaf proline dehydrogenase activity and significantly (P < 0.05) decreased leaf both Δ1-pyrroline-5-carboxylate reductase and Δ1-pyrroline-5-carboxylate synthetase activity, resulting in lower proline accumulation in AMF plants under DS. Our results therefore suggest that AMF strongly altered leaf sucrose and proline metabolism through regulating sucrose- and proline-metabolized enzyme activities, which is important for osmotic adjustment of the host plant. PMID:28181575

  7. Proline biosynthesis genes and their regulation under salinity stress in the euryhaline copepod Tigriopus californicus.

    Willett, Christopher S; Burton, Ronald S


    Diverse organisms regulate concentrations of intracellular organic osmolytes in response to changes in environmental salinity or desiccation. In marine crustaceans, accumulation of high concentrations of proline is a dominant component of response to hyperosmotic stress. In the euryhaline copepod Tigriopus californicus, synthesis of proline from its metabolic precursor glutamate is tightly regulated by changes in environmental salinity. Here, for the first time in a marine invertebrate, the genes responsible for this pathway have been cloned and characterized. The two proteins display the sequence features of homologous enzymes identified from other eukaryotes. One of the cloned genes, delta1-pyrroline-5-carboxylase reductase (P5CR), is demonstrated to have the reductase enzyme activity when expressed in proline-auxotroph bacteria, while the second, delta1-pyrroline-5-carboxylase synthase (P5CS), does not rescue proline-auxotroph bacteria. In contrast to results from higher plants, neither levels of P5CS nor P5CR mRNAs increase in response to salinity stress in T. californicus. Hence, regulation of proline synthesis during osmotic stress in T. californicus is likely mediated by some form of post-transcriptional regulation of either P5CS or P5CR. Understanding the regulation this pathway may elucidate the mechanisms limiting the salinity ranges of marine taxa.

  8. Changes in antioxidant enzymes activities and proline, total phenol and anthocyanine contents in Hyssopus officinalis L. plants under salt stress.

    Jahantigh, Omolbanin; Najafi, Farzaneh; Badi, Hassanali Naghdi; Khavari-Nejad, Ramazan Ali; Sanjarian, Forough


    The relationships between salt stress and antioxidant enzymes activities, proline, phenol and anthocyanine contents in Hyssopus officinalis L. plants in growth stage were investigated. The plants were subjected to five levels of saline irrigation water, 0.37 (tap water as control) with 2, 4, 6, 8 and 10 dSm(-1) of saline water. After two months the uniform plants were harvested for experimental analysis. Antioxidant enzymes activities and proline, phenol and anthocyanine contents of the plants were examinated. Enhanced activities of peroxidase, catalase and superoxide dismutase were determined by increasing salinity that plays an important protective role in the ROS-scavenging process. Proline, phenol and anthocyanine contents increased significantly with increasing salinity. These results suggest that salinity tolerance of Hyssopus officinalis plants might be closely related with the increased capacity of antioxidative system to scavenge reactive oxygen species and with the accumulation of osmoprotectant proline, phenol and anthocyanine contents under salinity conditions.

  9. The choice of the mathematical method for prediction of electrochemical accumulator parameters value in power installations of space-rocket objects

    Bezruchko, K. V.; Davidov, A. O.; Katorgina, J. G.; Logvin, V. M.; Kharchenko, A. A.


    The review and analysis of several mathematical methods for prediction of electrochemical accumulator parameters are provided in the article: according to the mathematical expectation, the latest entry, a statistical prediction, Box-Jenkins model, decomposition Volta, ARMA, ARIMA and Kalman filter. The results of these methods for prediction of the electrochemical battery 22НКГ-4CK characteristics which is a part of spacecraft power plant of the “Mikrosputnik” type are given. Possible usage of these methods for long prediction of electrochemical accumulator characteristics on space-rocket objects power plants is showed.

  10. Cadmium Induced Changes of Proline in Two Ecotypes of Thlaspi Caerulescens

    Zemanová V.


    Full Text Available A Thlaspi caerulescens (J. & C. PRESL was used to study the effect of cadmium on the content of free amino acids and ability accumulation of Cd in ecotypes of this plant species. In pot experiment two ecotypes T. caerulescens were used: Ganges ecotype from France and Mežica ecotype from Slovenia. The plants were grown in soil (chernozem – Suchdol spiked with NPK and three different concentration of Cd: 30, 60 and 90 mg/kg. The content of Cd was measured in the above-ground biomass and roots using ICP-OES. Accumulation of Cd was higher in the Mežica ecotype in contrast to the low Cd-accumulating the Ganges ecotype. Analyses of free amino acids contents were measured by GC-MS method. The content of free amino acids in above-ground biomass of the Mežica ecotype declined progressively with increasing concentrations of Cd. Opposite trend was observed in roots of this ecotype. The increase of free amino acids contents in above-ground biomass and roots of the Ganges ecotype were detected. The results of specific amino acids free proline showed increased content in plant biomass with increasing Cd contamination of soil. A statistically significant increase was observed between control plants (0 mg/kg Cd and variant Cd3 (90 mg/kg Cd for both ecotypes. The statistically significant decrease of free proline was observed in the Mežica ecotype roots. Opposite trend was observed in roots of Ganges ecotype - increasing trend of free proline content. These results indicate a correlation between content of Cd and content of free proline in different parts of the plant. We can speculate that the mechanism of Cd hyperaccumulation and metabolism of free proline are not identical in ecotypes of this species.

  11. Application of essential oils in maize grain: impact on Aspergillus section Flavi growth parameters and aflatoxin accumulation.

    Bluma, Romina V; Etcheverry, Miriam G


    The antifungal activity of Pimpinella anisum L. (anise), Pëumus boldus Mol (boldus), Hedeoma multiflora Benth (mountain thyme), Syzygium aromaticum L. (clove), and Lippia turbinate var. integrifolia (griseb) (poleo) essential oils (EOs) against Aspergillus section Flavi was evaluated in sterile maize grain under different water activity (a(w)) condition (0.982, 0.955, and 0.90). The effect of EOs added to maize grains on growth rate, lag phase, and aflatoxin B(1) (AFB(1)) accumulation of Aspergillus section Flavi were evaluated at different water activity conditions. The five EOs analyzed have been shown to influence lag phase and growth rate. Their efficacy depended mainly on the essential oil concentrations and substrate water activity conditions. All EOs showed significant impact on AFB(1) accumulation. This effect was closely dependent on the water activity, concentration, and incubation periods. Important reduction of AFB(1) accumulation was observed in the majority of EO treatments at 11 days of incubation. Boldus, poleo, and mountain thyme EO completely inhibited AFB(1) at 2000 and 3000 microg g(-1). Inhibition of AFB(1) accumulation was also observed when aflatoxigenic isolates grew with different concentration of EOs during 35 days.

  12. Conformational preferences of substituted prolines in the collagen triple helix.

    Mooney, Sean D; Kollman, Peter A; Klein, Teri E


    Researchers have recently questioned the role hydroxylated prolines play in stabilizing the collagen triple helix. To address these issues, we have developed new molecular mechanics parameters for the simulation of peptides containing 4(R)-fluoroproline (Flp), 4(R)-hydroxyproline (Hyp), and 4(R)-aminoproline (Amp). Simulations of peptides based on these parameters can be used to determine the components that stabilize hydroxyproline over proline in the triple helix. The dihedrals F-C-C-N, O-C-C-N, and N-C-C-N were built using a N-beta-ethyl amide model. One nanosecond simulations were performed on the trimers [(Pro-Pro-Gly)(10)](3), [(Pro-Hyp-Gly)(10)](3), [(Pro-Amp-Gly)(10)](3), [(Pro-Amp(1+)-Gly)(10)](3), and [(Pro-Flp-Gly)(10)](3) in explicit solvent. The results of our simulations suggest that pyrrolidine ring conformation is mediated by the strength of the gauche effect and classical electrostatic interactions.

  13. Proline synthesis in fat body of Leptinotarsa decemlineata

    Weeda, E.; Koopmanschap, A.B.; Kort, C.A.D. de; Beenakkers, A.M.Th.


    Fat body isolated from the Colorado potato beetle is capable of synthesizing proline. Maximum rate of proline synthesis is achieved with alanine as substrate. A metabolic pathway in which stored lipid participates in proline synthesis is suggested. The close relationship between flight muscle and fa

  14. Tertiary alcohol preferred: Hydroxylation of trans-3-methyl-L-proline with proline hydroxylases

    Christian Klein


    Full Text Available The enzymatic synthesis of tertiary alcohols by the stereospecific oxidation of tertiary alkyl centers is a most-straightforward but challenging approach, since these positions are sterically hindered. In contrast to P450-monooxygenases, there is little known about the potential of non-heme iron(II oxygenases to catalyze such reactions. We have studied the hydroxylation of trans-3-methyl-L-proline with the α-ketoglutarate (α-KG dependent oxygenases, cis-3-proline hydroxylase type II and cis-4-proline hydroxylase (cis-P3H_II and cis-P4H. With cis-P3H_II, the tertiary alcohol product (3R-3-hydroxy-3-methyl-L-proline was obtained exclusively but in reduced yield (~7% compared to the native substrate L-proline. For cis-P4H, a complete shift in regioselectivity from C-4 to C-3 was observed so that the same product as with cis-P3H_II was obtained. Moreover, the yields were at least as good as in control reactions with L-proline (~110% relative yield. This result demonstrates a remarkable potential of non-heme iron(II oxygenases to oxidize substrates selectively at sterically hindered positions.

  15. Physical parameters and accumulation rates in peat in relation to the climate during the last 150 years

    Borgmark, Anders [Dept. of Physical Geography and Quaternary Geology, Stockholm Univ. (Sweden)


    The safety assessment made by SKB (SR 97) states that radionuclides can be accumulated in higher amounts in peatlands than in other recipients. Therefore is knowledge about the nature and properties of peat very important. Here is the decay of peat and the accumulation rate of the most important elements of peat examined further. Two ombrotrophic peat bogs located in Uppland have been investigated in order to evaluate the influences of climate on the accumulation of carbon and nitrogen. Peat humification, content of carbon and nitrogen has been used for interpretation of peat forming processes. The long temperature and precipitation records from Uppsala have been used to compare the results to known climate variations. Various models and equations assess the contribution of peatlands to the global carbon economy and the role that peat accumulation plays in global climate due to global carbon cycling and the concern of increasing levels of greenhouse gases. The importance of peatlands in the global carbon economy is stressed by that it is approximately the same amount of carbon in peatlands as in the atmosphere. Estimations of the total amount of carbon stored in Boreal and Arctic peatlands are in the magnitude of 400-500 Pg. The peat accumulation rate varies by at least a magnitude in peatlands with different conditions in internal and external hydrology, length in growth season, effective precipitation, temperature etc. Accumulation rates have been reported from a variety of temperate and boreal bogs ranging between 0.2-2.0 mm/year and a Boreal and northern Sub-arctic region average of carbon accumulated in the catotelm has been calculated to ca 21 g C/m{sup 2}/year. The proportion of nitrogen in the dry mass is usually in the order of 0.5-5%. The mean accumulation rate of carbon and nitrogen during the last 157 years at Aeltabergsmossen are 7 g C/m{sup 2}/year and 1 g N/m{sup 2}/year, these levels are similar to the ones found in other investigations as well as

  16. Evolution of proline biosynthesis: enzymology, bioinformatics, genetics, and transcriptional regulation.

    Fichman, Yosef; Gerdes, Svetlana Y; Kovács, Hajnalka; Szabados, László; Zilberstein, Aviah; Csonka, Laszlo N


    Proline is not only an essential component of proteins but it also has important roles in adaptation to osmotic and dehydration stresses, redox control, and apoptosis. Here, we review pathways of proline biosynthesis in the three domains of life. Pathway reconstruction from genome data for hundreds of eubacterial and dozens of archaeal and eukaryotic organisms revealed evolutionary conservation and variations of this pathway across different taxa. In the most prevalent pathway of proline synthesis, glutamate is phosphorylated to γ-glutamyl phosphate by γ-glutamyl kinase, reduced to γ-glutamyl semialdehyde by γ-glutamyl phosphate reductase, cyclized spontaneously to Δ(1)-pyrroline-5-carboxylate and reduced to proline by Δ(1)-pyrroline-5-carboxylate reductase. In higher plants and animals the first two steps are catalysed by a bi-functional Δ(1) -pyrroline-5-carboxylate synthase. Alternative pathways of proline formation use the initial steps of the arginine biosynthetic pathway to ornithine, which can be converted to Δ(1)-pyrroline-5-carboxylate by ornithine aminotransferase and then reduced to proline or converted directly to proline by ornithine cyclodeaminase. In some organisms, the latter pathways contribute to or could be fully responsible for the synthesis of proline. The conservation of proline biosynthetic enzymes and significance of specific residues for catalytic activity and allosteric regulation are analysed on the basis of protein structural data, multiple sequence alignments, and mutant studies, providing novel insights into proline biosynthesis in organisms. We also discuss the transcriptional control of the proline biosynthetic genes in bacteria and plants.

  17. Rapid accumulation and low degradation: key parameters of Tomato yellow leaf curl virus persistence in its insect vector Bemisia tabaci.

    Becker, Nathalie; Rimbaud, Loup; Chiroleu, Frédéric; Reynaud, Bernard; Thébaud, Gaël; Lett, Jean-Michel


    Of worldwide economic importance, Tomato yellow leaf curl virus (TYLCV, Begomovirus) is responsible for one of the most devastating plant diseases in warm and temperate regions. The DNA begomoviruses (Geminiviridae) are transmitted by the whitefly species complex Bemisia tabaci. Although geminiviruses have long been described as circulative non-propagative viruses, observations such as long persistence of TYLCV in B. tabaci raised the question of their possible replication in the vector. We monitored two major TYLCV strains, Mild (Mld) and Israel (IL), in the invasive B. tabaci Middle East-Asia Minor 1 cryptic species, during and after the viral acquisition, within two timeframes (0-144 hours or 0-20 days). TYLCV DNA was quantified using real-time PCR, and the complementary DNA strand of TYLCV involved in viral replication was specifically quantified using anchored real-time PCR. The DNA of both TYLCV strains accumulated exponentially during acquisition but remained stable after viral acquisition had stopped. Neither replication nor vertical transmission were observed. In conclusion, our quantification of the viral loads and complementary strands of both Mld and IL strains of TYLCV in B. tabaci point to an efficient accumulation and preservation mechanism, rather than to a dynamic equilibrium between replication and degradation.

  18. Variance of genetic parameters for gas accumulations in the Rotliegende of the Western Altmark; Varianz lagerstaettengenetischer Parameter im Rotliegenden der westlichen Altmark

    Schumacher, K.H. [Erdoel-Erdgas Gommern GmbH, Gommern (Germany); May, F. [Erdoel-Erdgas Gommern GmbH, Gommern (Germany)


    The gas fields of the western Altmark show a distinct zonal variance of gas- and isotope-geochemical and lithological parameters. The diapir-like distribution of methane contents and isotopes ratios gives information not only on the role played by the N-S faults associated with the Rotliegende rifting in providing gas-migration paths, but also on special aspects of the migration process. (orig.) [Deutsch] In den Lagerstaetten der westlichen Altmark tritt eine ausgepraegte zonale Varianz erdgas- und isotopengeochemischer sowie lithologischer Parameter auf. An Hand diapirartiger Erscheinungsbilder der Methangehalts- und Isotopenverteilungen werden neben der Rolle der N-S-gerichteten Bruchstoerungen des Rotliegend-Riftings fuer die Erdgaszufuhr zugleich Besonderheiten des Migrationsablaufes deutlich. (orig.)

  19. Genotype differences in the metabolism of proline and polyamines under moderate drought in tomato plants.

    Montesinos-Pereira, D; Barrameda-Medina, Y; Romero, L; Ruiz, J M; Sánchez-Rodríguez, E


    Water stress is one of the most important factors limiting the growth and productivity of crops. The implication of compatible osmolytes such as proline and polyamines in osmotic adjustment has been widely described in numerous plants species under stress conditions. In the present study, we investigated the response of five cherry tomato cultivars (Solanum lycopersicum L.) subjected to moderate water stress in order to shed light on the involvement of proline and polyamine metabolism in the mechanisms of tolerance to moderate water stress. Our results indicate that the most water stress-resistant cultivar (Zarina) had increased degradation of proline associated with increased polyamine synthesis, with a higher concentration of spermidine and spermine under stress conditions. In contrast, Josefina, the cultivar most sensitive to water stress, showed a proline accumulation associated with increased synthesis after being subjected to stress. In turn, in this cultivar, no rise in polyamine synthesis was detected. Therefore, all the data appear to indicate that polyamine metabolism is more involved in the tolerance response to moderate water stress.

  20. Variação nos níveis de prolina, diamina e poliaminas em cultivares de trigo submetidas a déficits hídricos Proline, diamine and polyamines accumulation in wheat cultivars submitted to water deficits

    Terezinha de Fátima Fumis


    Full Text Available O objetivo deste trabalho foi avaliar duas cultivares de trigo (Triticum aestivum L. Anahuac e IAC-24, cultivadas em solo arenoso, sob três regimes de umidade do solo (-0,027 a -0,1 MPa; -0,027 a -0,045 MPa; e -0,027 MPa -- irrigação constante. Foram analisadas as variações nos níveis de prolina, da diamina (putrescina e das poliaminas (espermina e espermidina e o teor relativo de água (TRA durante o perfilhamento, emborrachamento, floração e no enchimento dos grãos. Nas plantas submetidas a déficits hídricos foi verificada menor TRA e acúmulo de prolina e putrescina; em relação às poliaminas não foram observadas diferenças entre os tratamentos. Nos tratamentos sob déficit hídrico, a cultivar IAC-24 apresentou teores mais elevados de prolina e putrescina, o mesmo sendo verificado com a cultivar Anahuac quando submetida a tratamento sem déficit hídrico.The objective of this work was to evaluate two wheat (Triticum aestivum L. cultivars, Anahuac and IAC-24, developed in sandy texture soil, under three levels of soil humidity (-0.027 to -0.1 MPa; -0.027 to -0.045 MPa; and -0.027 MPa constantly irrigated. The variation in the levels of proline, diamine (putrescine and polyamines (spermine and spermidine content and the relative water content (RWC during the tillering, booting, flowering and grain-filling was determined. The plants under water deficits presented smaller RWC and higher levels of proline and putrescine. In relation to the polyamines it wasn't observed difference among the treatments. The cv. IAC-24 under water deficit presented a higher content of proline and putrescine while the Anahuac showed the same results in the treatments without water deficit.

  1. Effect of salinity stress on chlorophyll content, proline, water soluble carbohydrate, germination, growth and dry weight of three seedling barley (Hordeum vulgare L. cultivars

    Movafegh S.


    Full Text Available Salinity is a serious environmental constraint to crop production in many parts of the world and the development of crops with improved salt tolerance is proposed as part of solution to this problem. This research was performed out in order to study the effects of different salinity levels on germination, growth, dry weight, proline, water soluble carbohydrate and chlorophyll content of three barley (Hordeum vulgare L. cultivars named Jonoob (INC-54, Reyhan (INC-45 & Nosrat (INC -47.The experiment was carried out using factorial based on completely randomized design with three replications.Seven old seedlings after germination were transferred to Hoagland nutrient solution under the effect of salinity levels (0, 50, 150 and 250 mM NaClin during seven days. Data variance analysis showed that seed germination of three barley cultivars was significantly (PReyhan>Nosrat. The results showed that, increasing in salinity decreased all growth parameters. Salinity stress decreased shoot and root length, root dry weight and chlorophyll contents in every three cultivars. But decreasing of chlorophyll was less in Nosrat compared to two other items. Proline content and soluble carbohydrate were increased in all of the three cultivars with enhance of NaCl concentration. By increasing of salinity stress accumulation of proline and soluble sugar content in leaves of Nosrat cultivar was more than other cultivars.As saltiness increases resistance natural responses in this plant gets better considering less decrease in chlorophyll amount and strategy of more production about praline and sugar solution compared to two other items.

  2. Effect of MgCl2 stress on germination, plant growth, chlorophyll content, proline content and lipid peroxidation in sorghum cultivars

    Khare T.


    Full Text Available Present report deals with the effect of increasing levels (0-300 mM of MgCl2 salt on sorghum cultivars, Phule Vasudha and Phule Revati. Although MgCl2 stress did not show considerable adverse effects on germination, however, at higher (>200 mM concentrations, the seedlings turned brown and did not showed any further growth, with comparably higher magnitude of negative effects on Phule Revati than Phule Vasudha. Overall, increasing MgCl2 stress reduced plant growth and biomass production significantly in both the cultivars, though with lesser extent in Phule Vsudha as compared to Phule Rvati. Contrasting behavior was evidenced in both the cultivars in terms of protein content under varying levels of MgCl2 concentration at vegetative growth level, where salinity induced reduction in protein content was higher in Phule Revati than Phule Vasudha. The genotype Phule Vasudha showed higher proline content under non-saline condition. MgCl2 stress-induced proline accumulation was observed in both the sorghum cultivars, however, interestingly, Phule Revati (439% of control plants showed comparably higher proline content than Phule Vasudha (324% of control plants at the highest (300 mM level of stress. Even though, malondialdehyde (MDA: lipid peroxidation indicator content was on higher side under non-saline conditions in cultivar Phule Vasudha as compared to cultivar Phule Revati, however the rate of increase in MDA with increasing salt stress was much higher in the latter cultivar, indicating the comparably higher level of lipid peroxidation under the influence of MgCl2 stress. The salt tolerance nature of Phule Vasudha was positively correlated with its better performance in terms of physiological and biochemical parameters.

  3. L-Proline nutrition and catabolism in Staphylococcus saprophyticus.

    Deutch, Charles E


    Staphylococcus saprophyticus strains ATCC 15305, ATCC 35552, and ATCC 49907 were found to require L-proline but not L-arginine for growth in a defined culture medium. All three strains could utilize L-ornithine as a proline source and contained L-ornithine aminotransferase and Δ(1)-pyrroline-5-carboxylate reductase activities; strains ATCC 35552 and ATCC 49907 could use L-arginine as a proline source and had L-arginase activity. The proline requirement also could be met by L-prolinamide, L-proline methyl ester, and the dipeptides L-alanyl-L-proline and L-leucyl-L-proline. The bacteria exhibited L-proline degradative activity as measured by the formation of Δ(1)-pyrroline-5-carboxylate. The specific activity of proline degradation was not affected by addition of L-proline or NaCl but was highest in strain ATCC 49907 after growth in Mueller-Hinton broth. A membrane fraction from this strain had L-proline dehydrogenase activity as detected both by reaction of Δ(1)-pyrroline-5-carboxylate with 2-aminobenzaldehyde (0.79 nmol min(-1) mg(-1)) and by the proline-dependent reduction of p-iodonitrotetrazolium (20.1 nmol min(-1) mg(-1)). A soluble fraction from this strain had Δ(1)-pyrroline-5-carboxylate dehydrogenase activity (88.8 nmol min(-1) mg(-1)) as determined by the NAD(+)-dependent oxidation of DL-Δ(1)-pyrroline-5-carboxylate. Addition of L-proline to several culture media did not increase the growth rate or final yield of bacteria but did stimulate growth during osmotic stress. When grown with L: -ornithine as the proline source, S. saprophyticus was most susceptible to the proline analogues L-azetidine-2-carboylate, 3,4-dehydro-DL-proline, DL-thiazolidine-2-carboxylate, and L-thiazolidine-4-carboxylate. These results indicate that proline uptake and metabolism may be a potential target of antimicrobial therapy for this organism.

  4. SKN-1 and Nrf2 couples proline catabolism with lipid metabolism during nutrient deprivation.

    Pang, Shanshan; Lynn, Dana A; Lo, Jacqueline Y; Paek, Jennifer; Curran, Sean P


    Mechanisms that coordinate different metabolic pathways, such as glucose and lipid, have been recognized. However, a potential interaction between amino acid and lipid metabolism remains largely elusive. Here we show that during starvation of Caenorhabditis elegans, proline catabolism is coupled with lipid metabolism by SKN-1. Mutation of alh-6, a conserved proline catabolic enzyme, accelerates fat mobilization, enhances the expression of genes involved in fatty acid oxidation and reduces survival in response to fasting. This metabolic coordination is mediated by the activation of the transcription factor SKN-1/Nrf2, possibly due to the accumulation of the alh-6 substrate P5C, and also requires the transcriptional co-regulator MDT-15. Constitutive activation of SKN-1 induces a similar transcriptional response, which protects animals from fat accumulation when fed a high carbohydrate diet. In human cells, an orthologous alh-6 enzyme, ALDH4A1, is also linked to the activity of Nrf2, the human orthologue of SKN-1, and regulates the expression of lipid metabolic genes. Our findings identify a link between proline catabolism and lipid metabolism, and uncover a physiological role for SKN-1 in metabolism.

  5. Influence of some soil parameters on heavy metals accumulation by vegetables grown in agricultural soils of different soil orders.

    Golia, E E; Dimirkou, A; Mitsios, I K


    The main purpose of this research was to determine the levels of heavy metals in tomato, potato and lettuce, grown in agricultural soils of different soil orders (Alfisols, Endisols and Vertisols), located at Central Greece. Soil samples were analysed for available forms (after extraction with DTPA) and for total concentrations (after digestion with Aqua Regia) of metals. Zn, Cu, Cr and Ni were the common metals detected in the vegetables studied. Pb and Cd concentrations were low and in some cases not detectable. Significant correlations among metals concentrations and soil physicochemical parameters were obtained and discussed. The pH value and the percentage of clay content were found to determine the solubility of metals in the soil and their availability for uptake by plants.

  6. Connecting proline metabolism and signaling pathways in plant senescence

    Lu eZhang


    Full Text Available The amino acid proline has a unique biological role in stress adaptation. Proline metabolism is manipulated under stress by multiple and complex regulatory pathways and can profoundly influence cell death and survival in microorganisms, plants, and animals. Though the effects of proline are mediated by diverse signaling pathways, a common theme appears to be the generation of reactive oxygen species (ROS due to proline oxidation being coupled to the respiratory electron transport chain. Considerable research has been devoted to understand how plants exploit proline metabolism in response to abiotic and biotic stress. Here, we review potential mechanisms by which proline metabolism influences plant senescence, namely in the petal and leaf. Recent studies of petal senescence suggest proline content is manipulated to meet energy demands of senescing cells. In the flower and leaf, proline metabolism may influence ROS signaling pathways that delay senescence progression. Future studies focusing on the mechanisms by which proline metabolic shifts occur during senescence may lead to novel methods to rescue crops under stress and to preserve post-harvest agricultural products.

  7. Proline and Abscisic Acid Content in Droughted Corn Plant Inoculated with Azospirillum sp. and Arbuscular Mycorrhizae Fungi



    Full Text Available Plants that undergo drought stress perform a physiological response such as accumulation of proline in the leaves and increased content abscisic acid. A research was conducted to study proline and abscisic acid (ABA content on drought-stressed corn plant with Azospirillum sp. and arbuscular mycorrhizae fungi (AMF inoculated at inceptisol soil from Bogor, West Java. The experiments were carried out in a green house from June up to September 2003, using a factorial randomized block design. In pot experiments, two factors were assigned, i.e. inoculation with Azospirillum (0, 0.50, 1.00, 1.50 ml/pot and inoculation with AMF Glomus manihotis (0, 12.50, 25.00, 37.50 g/pot. The plants were observed during tasseling up to seed filling periods. Results of experiments showed that the interaction between Azospirillum sp. and AMF was synergistically increased proline, however it decreased ABA.

  8. Molecular cloning and characterization of a gene encoding the proline transporter protein in common bean (Phaseolus vulgaris L.

    Jibao Chen


    Full Text Available As a typical compatible solute, proline is accumulated in plants under environmental stresses. Proline transporter (ProT plays an important role in proline distribution between plant organs. Using a candidate gene approach, we cloned a cDNA sequence for ProT from common bean (Phaseolus vulgaris L. and designated the gene PvProT. The deduced amino acid sequence of PvProT showed high similarity to Bet/ProT proteins from other leguminous plants, and the highest similarity was observed with mothbean (Vigna aconitifolia L. VuProT. Relative quantification of the mRNA level of PvProT using real-time PCR analysis showed that the PvProT transcript level was higher in leaves than in stems and roots of common bean plants subjected to drought and salt stress. Under 20% (w/w PEG-6000 treatment, drought-resistant plants expressed a higher level of PvProT transcripts than drought-sensitive plants. Although heterologous expression of PvProT in the Escherichia coli mutant mkh13 showed that PvProT exhibited uptake activities for proline and betaine, no betaine content was detected in the common bean. These findings suggest that PvProT plays an important role in the transportation of proline in common bean plants exposed to drought and salt stress.

  9. Molecular cloning and characterization of a gene encoding the proline transporter protein in common bean (Phaseolus vulgaris L.)

    Jibao Chen; Jing Wu; Yunfeng Lu; Yuannan Cao; Hui Zeng; Zhaoyuan Zhang; Lanfen Wang; Shumin Wang


    As a typical compatible solute, proline is accumulated in plants under environmental stresses. Proline transporter (ProT) plays an important role in proline distribution between plant organs. Using a candidate gene approach, we cloned a cDNA sequence for ProT from common bean (Phaseolus vulgaris L.) and designated the gene PvProT. The deduced amino acid sequence of PvProT showed high similarity to Bet/ProT proteins from other leguminous plants, and the highest similarity was observed with mothbean (Vigna aconitifolia L.) VuProT. Relative quantification of the mRNA level of PvProT using real-time PCR analysis showed that the PvProT transcript level was higher in leaves than in stems and roots of common bean plants subjected to drought and salt stress. Under 20% (w/w) PEG-6000 treatment, drought-resistant plants expressed a higher level of PvProT transcripts than drought-sensitive plants. Although heterologous expression of PvProT in the Escherichia coli mutant mkh13 showed that PvProT exhibited uptake activities for proline and betaine, no betaine content was detected in the common bean. These findings suggest that PvProT plays an important role in the transportation of proline in common bean plants exposed to drought and salt stress.

  10. Stereoretentive formylation of (S)-proline

    Temizsoy, Mehmet; Sethi, Waqas; Reinholdt, Anders


    In a Vilsmeier-Haack-type formylation reaction the α-(dihydroxymethyl)-(S)-prolinato complex (+)578-p-[Co(tren){(RC,SN)-Pro[CH(OH)2]O}]Cl2·2H2O (22) was produced stereoselectively (85% ee) from the (S)-prolinato complex, (+)578-p-[Co(tren){(SC,SN)-ProO}]2(H3O)2(HOEt2)(O3SCF3)7 (18). Similar...... reaction of the (S)-alaninato complex, (-)578-p-[Co(tren)(S-AlaO)](H3O)(O3SCF3)3 (13), produced the racemate, rac-p-[Co(tren)(Ala{CH(OH)2}O)]SO4·2H2O (17). The contrasting stereochemical outcomes of the formylation reaction with 18versus13 were ascribed to the stereogenic character of the coordinated sec....... amine of the (S)-prolinate chelate in 18, which serves to uphold a chiral environment during reaction, whereas reaction intermediates derived from 13 lack this stereochemical feature. The stereoselective formylation of (S)-proline, relying on coordination to an inert metal centre, as conducted here...

  11. Changes of Proline Content,Activity,and Active Isoforms of Antioxidative Enzymes in Two Alfalfa Cultivars Under Salt Stress

    WANG Xiao-shan; HAN Jian-guo


    The plants of two alfalfa(Medicago sativa L.)cultivars differing in salt tolerance were subjected to three salt treatments,70,140,and 210 mM NaCl for 7 days.Root,shoot,and leaf growths were inhibited by increased salt treatments in both cultivars,and at 140 and 210 mM salt treatments,Zhongmu 1 had significantly higher root,shoot,and leaf dry weights per plant than Defi.The malondialdehyde(MDA)accumulation in Defi was considerably greater than in Zhongmu 1,indicating a higher degree of lipid peroxidation at 140 and 210 mM salt treatments.The changes in the activity and active isoforms of antioxidant enzymes such as superoxide dismutase(SOD,EC,catalase(CAT,EC,peroxidase(POD,EC,and ascorbate peroxidase(APOX,EC,accumulation of free proline,and rate of lipid peroxidation in leaves of two alfalfa cultivars were also investigated.After stress,the activity and active isoforms of antioxidative enzymes were altered and the extent of alteration varied between the cultivar Deft and Zhongmu 1.The proline accumulation in Defi was considerably greater than in Zhongmu 1 at 210 mM salt treatment.This indicated that proline accumulation may be the result,instead of the cause,of salt tolerance.

  12. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to isoleucine-proline-proline (IPP) and valine-proline-proline (VPP) and maintenance of normal blood pressure (ID 661, 1831, 1832, 2891, further assessment) pursuant

    Tetens, Inge

    Following a request from the European Commission, pursuant to Article 13.1 of Regulation (EC) No 1924/2006, the Panel on Dietetic Products, Nutrition and Allergies was asked to provide a scientific opinion on a health claim related to isoleucine-proline-proline (IPP) and valine-proline-proline (VPP......) and maintenance of normal blood pressure. The food constituent that is the subject of the claim, the tripeptides isoleucine-proline-proline (IPP) and valine-proline-proline (VPP), is sufficiently characterised. The claimed effect, maintenance of normal blood pressure, is a beneficial physiological effect...

  13. Roles of conserved proline and glycosyltransferase motifs of EmbC in biosynthesis of lipoarabinomannan.

    Berg, Stefan; Starbuck, James; Torrelles, Jordi B; Vissa, Varalakshmi D; Crick, Dean C; Chatterjee, Delphi; Brennan, Patrick J


    D-Arabinans, composed of D-arabinofuranose (D-Araf), dominate the structure of mycobacterial cell walls in two settings, as part of lipoarabinomannan (LAM) and arabinogalactan, each with markedly different structures and functions. Little is known of the complexity of their biosynthesis. beta-D-Arabinofuranosyl-1-monophosphoryldecaprenol is the only known sugar donor. EmbA, EmbB, and EmbC, products of the paralogous genes embA, embB, and embC, the sites of resistance to the anti-tuberculosis drug ethambutol (EMB), are the only known implicated enzymes. EmbA and -B apparently contribute to the synthesis of arabinogalactan, whereas EmbC is reserved for the synthesis of LAM. The Emb proteins show no overall similarity to any known proteins beyond Mycobacterium and related genera. However, functional motifs, equivalent to a proline-rich motif of several bacterial polysaccharide co-polymerases and a superfamily of glycosyltransferases, were found. Site-directed mutagenesis in glycosyltransferase superfamily C resulted in complete ablation of LAM synthesis. Point mutations in three amino acids of the proline motif of EmbC resulted in marked reduction of LAM-arabinan synthesis and accumulation of an unknown intermediate and of the known precursor lipomannan. Yet the pattern of the differently linked d-Araf units observed in wild type LAM-arabinan was largely retained in the proline motif mutants. The results allow for the presentation of a unique model of arabinan synthesis.

  14. β-amylase 1 (BAM1) degrades transitory starch to sustain proline biosynthesis during drought stress.

    Zanella, Martina; Borghi, Gian Luca; Pirone, Claudia; Thalmann, Matthias; Pazmino, Diana; Costa, Alex; Santelia, Diana; Trost, Paolo; Sparla, Francesca


    During photosynthesis of higher plants, absorbed light energy is converted into chemical energy that, in part, is accumulated in the form of transitory starch within chloroplasts. In the following night, transitory starch is mobilized to sustain the heterotrophic metabolism of the plant. β-amylases are glucan hydrolases that cleave α-1,4-glycosidic bonds of starch and release maltose units from the non-reducing end of the polysaccharide chain. In Arabidopsis, nocturnal degradation of transitory starch involves mainly β-amylase-3 (BAM3). A second β-amylase isoform, β-amylase-1 (BAM1), is involved in diurnal starch degradation in guard cells, a process that sustains stomata opening. However, BAM1 also contributes to diurnal starch turnover in mesophyll cells under osmotic stress. With the aim of dissecting the role of β-amylases in osmotic stress responses in Arabidopsis, mutant plants lacking either BAM1 or BAM3 were subject to a mild (150mM mannitol) and prolonged (up to one week) osmotic stress. We show here that leaves of osmotically-stressed bam1 plants accumulated more starch and fewer soluble sugars than both wild-type and bam3 plants during the day. Moreover, bam1 mutants were impaired in proline accumulation and suffered from stronger lipid peroxidation, compared with both wild-type and bam3 plants. Taken together, these data strongly suggest that carbon skeletons deriving from BAM1 diurnal degradation of transitory starch support the biosynthesis of proline required to face the osmotic stress. We propose the transitory-starch/proline interplay as an interesting trait to be tackled by breeding technologies aimingto improve drought tolerance in relevant crops.

  15. Effects of three stabilizing agents--proline, betaine, and trehalose--on membrane phospholipids.

    Rudolph, A S; Crowe, J H; Crowe, L M


    We have studied the interaction between three compounds which accumulate in organisms under hydration stress--proline, betaine, and trehalose--and the membrane phospholipids dimyristoylphosphatidylcholine (DMPC), palmitoyloleoylphosphatidylcholine (POPC), and dimyristoylphosphatidylethanolamine in bulk solution. Film balance studies reveal that these compounds increase the area/molecule of these lipids. Differential scanning calorimetry has been employed to investigate the effect these agents have on the gel-to-liquid crystalline phase transition of multilamellar and small unilamellar vesicles of DMPC, dipalmitoylphosphatidylcholine, and POPC:phosphatidylserine (90:10 mole ratio) in bulk solution. In the presence of 1 M proline, trehalose, or betaine, the midtransition temperature in small unilamellar vesicles is reduced (up to 7 degrees C in 1 M trehalose), and the transition broadened. In contrast, multilamellar vesicles of similar lipid composition show an increased transition temperature in the presence of the same concentration of these compounds. This result suggests that the inner lamellae in multilamellar vesicles may be dehydrated with only a few outer lamellae exposed to the protective compound. Finally, we have used stereomodels of phosphatidylcholine to investigate the mechanism of action of these agents. Hydrogen bonding of trehalose to the head group region results in an increase in the distance between head groups of 6.9 A. This amount of spreading compares well with data from the monolayer experiments which indicate that maximal spreading of DMPC monolayers by trehalose is 6.5 A. Molecular models of proline and betaine have also been constructed, and these models suggest potential interactions between these compounds and phosphatidylcholines. For the amphipath proline, this interaction may involve intercalation between phospholipid head groups.

  16. Quantifying additive interactions of the osmolyte proline with individual functional groups of proteins: comparisons with urea and glycine betaine, interpretation of m-values.

    Diehl, Roger C; Guinn, Emily J; Capp, Michael W; Tsodikov, Oleg V; Record, M Thomas


    To quantify interactions of the osmolyte l-proline with protein functional groups and predict their effects on protein processes, we use vapor pressure osmometry to determine chemical potential derivatives dμ2/dm3 = μ23, quantifying the preferential interactions of proline (component 3) with 21 solutes (component 2) selected to display different combinations of aliphatic or aromatic C, amide, carboxylate, phosphate or hydroxyl O, and amide or cationic N surface. Solubility data yield μ23 values for four less-soluble solutes. Values of μ23 are dissected using an ASA-based analysis to test the hypothesis of additivity and obtain α-values (proline interaction potentials) for these eight surface types and three inorganic ions. Values of μ23 predicted from these α-values agree with the experiment, demonstrating additivity. Molecular interpretation of α-values using the solute partitioning model yields partition coefficients (Kp) quantifying the local accumulation or exclusion of proline in the hydration water of each functional group. Interactions of proline with native protein surfaces and effects of proline on protein unfolding are predicted from α-values and ASA information and compared with experimental data, with results for glycine betaine and urea, and with predictions from transfer free energy analysis. We conclude that proline stabilizes proteins because of its unfavorable interactions with (exclusion from) amide oxygens and aliphatic hydrocarbon surfaces exposed in unfolding and that proline is an effective in vivo osmolyte because of the osmolality increase resulting from its unfavorable interactions with anionic (carboxylate and phosphate) and amide oxygens and aliphatic hydrocarbon groups on the surface of cytoplasmic proteins and nucleic acids.

  17. Multicomponent asymmetric reactions mediated by proline lithium salt

    Renzi, Polyssena; Overgaard, Jacob; Bella, Marco


    The multicomponent reaction between proline lithium salt, 2-cyclohexen-1-one and aliphatic aldehydes affords the 4- alkylidene-2-cyclohexen-1-ones, which are interesting fragrances, and bicyclic amino acids that bear four additional stereocenters, obtained as single stereoisomer....

  18. Proline Coordination with Fatty Acid Synthesis and Redox Metabolism of Chloroplast and Mitochondria.

    Shinde, Suhas; Villamor, Joji Grace; Lin, Wendar; Sharma, Sandeep; Verslues, Paul E


    Proline (Pro) accumulation is one of the most prominent changes in plant metabolism during drought and low water potential; however, the regulation and function of Pro metabolism remain unclear. We used a combination of forward genetic screening based on a Proline Dehydrogenase1 (PDH1) promoter-luciferase reporter (PDH1pro:LUC2) and RNA sequencing of the Pro synthesis mutant p5cs1-4 to identify multiple loci affecting Pro accumulation in Arabidopsis (Arabidopsis thaliana). Two mutants having high PDH1pro:LUC2 expression and increased Pro accumulation at low water potential were found to be alleles of Cytochrome P450, Family 86, Subfamily A, Polypeptide2 (CYP86A2) and Long Chain Acyl Synthetase2 (LACS2), which catalyze two successive steps in very-long-chain fatty acid (VLCFA) synthesis. Reverse genetic experiments found additional VLCFA and lipid metabolism-related mutants with increased Pro accumulation. Altered cellular redox status is a key factor in the coordination of Pro and VLCFA metabolism. The NADPH oxidase inhibitor diphenyleneiodonium (DPI) induced high levels of Pro accumulation and strongly repressed PDH1pro:LUC2 expression. cyp86a2 and lacs2 mutants were hypersensitive to diphenyleneiodonium but could be reverted to wild-type Pro and PDH1pro:LUC2 expression by reactive oxygen species scavengers. The coordination of Pro and redox metabolism also was indicated by the altered expression of chloroplast and mitochondria electron transport genes in p5cs1-4 These results show that Pro metabolism is both influenced by and influences cellular redox status via previously unknown coordination with several metabolic pathways. In particular, Pro and VLCFA synthesis share dual roles to help buffer cellular redox status while producing products useful for stress resistance, namely the compatible solute Pro and cuticle lipids. © 2016 American Society of Plant Biologists. All Rights Reserved.

  19. Comparative aspects of tissue glutamine and proline metabolism.

    Bertolo, Robert F; Burrin, Douglas G


    The cellular metabolism of glutamine and proline are closely interrelated, because they can be interconverted with glutamate and ornithine via the mitochondrial pathway involving pyrroline-5-carboxylate (P5C). In adults, glutamine and proline are converted via P5C to citrulline in the gut, then citrulline is converted to arginine in the kidney. In neonates, arginine is a semiindispensable amino acid and is synthesized from proline completely in the gut; because of low P5C synthase activity, glutamine is not an important precursor for neonatal arginine synthesis. Thus, splanchnic metabolism of glutamine and proline is important, because both amino acids serve as key precursors for arginine synthesis with some developmental differences. Studies investigating splanchnic extraction demonstrate that about two-thirds of dietary glutamine and almost all dietary glutamate are extracted on first pass and the vast majority is oxidized in the gut. This capacity to extract glutamine and glutamate appears to be very large, so diets high in glutamine or glutamate probably have little impact on circulating concentrations and consequent potential toxicity. In contrast, it appears that very little proline is extracted by the gut and liver, at least in the neonate, which may result in hyperprolinemia and potential toxicity. Therefore, the upper limits of safe dietary intake for glutamine and proline, and other amino acids, appear to be substantially different depending on the extent of first-pass splanchnic extraction and irreversible catabolism.

  20. Cigarette smoke enhances chemotaxis via acetylation of proline-glycine-proline.

    Hardison, Matthew Thomas; Brown, Michael David; Snelgrove, Robert James; Blalock, James Edwin; Jackson, Patricia


    Several chronic lung diseases have been linked to cigarette smoking (Chronic Obstructive Pulmonary Disease (COPD), and cancer are associated with increased tobacco use). We recently described a collagen fragment, proline-glycine-proline (PGP), chemotactic for neutrophils, that appears to play a role in COPD, cystic fibrosis, and bronchiolitis obliterans syndrome. PGP can exist in either its native or acetylated form (NAcPGP), although the mechanism of N-terminal-acetylation remains unknown. This work investigates the possibility that cigarette smoke (CS) and its components acetylate PGP, describing a possible mechanism for some of the chronic inflammation seen in tobacco-associated disease. CSE and CSC (3.56 and 12.38 ng/ml NAcPGP respectively, p less than 0.01) and its components (acrolein, acetaldehyde, and methyl glyoxal) acetylated PGP (0.51, 1.03, and 0.23 ng/ml NAcPGP, p less than 0.01). Both N-acetyl-cysteine and carbocysteine (scavengers of reactive aldehydes) blocked chemical acetylation of PGP by CS (100 percent and 97 percent inhibition, respectively, p less than 0.01). NAcPGP is more chemoattractive to neutrophils, and less susceptible to degradation by Leukotriene-A4-Hydrolase (detected in the lung). These experiments propose a mechanism for the increased neutrophil recruitment seen in smoking-associated lung diseases.

  1. Enhanced expression of the proline synthesis gene P5CSA in relation to seed osmopriming improvement of Brassica napus germination under salinity stress.

    Kubala, Szymon; Wojtyla, Łukasz; Quinet, Muriel; Lechowska, Katarzyna; Lutts, Stanley; Garnczarska, Małgorzata


    Osmopriming is a pre-sowing treatment that enhances germination performance and stress tolerance of germinating seeds. Brassica napus seeds showed osmopriming-improved germination and seedling growth under salinity stress. To understand the molecular and biochemical mechanisms of osmopriming-induced salinity tolerance, the accumulation of proline, gene expression and activity of enzymes involved in proline metabolism and the level of endogenous hydrogen peroxide were investigated in rape seeds during osmopriming and post-priming germination under control (H2O) and stress conditions (100 mM NaCl). The relationship between gene expression and enzymatic activity of pyrroline-5-carboxylate synthetase (P5CS), ornithine-δ-aminotransferase (OAT) and proline dehydrogenase (PDH) was determined. The improved germination performance of osmoprimed seeds was accompanied by a significant increase in proline content. The accumulation of proline during priming and post-priming germination was associated with strong up-regulation of the P5CSA gene, down-regulation of the PDH gene and accumulation of hydrogen peroxide. The up-regulated transcript level of P5CSA was consistent with the increase in P5CS activity. This study shows, for the first time, the role of priming-induced modulation of activities of particular genes and enzymes of proline turnover, and its relationship with higher content of hydrogen peroxide, in improving seed germination under salinity stress. Following initial stress-exposure, the primed seeds acquired stronger salinity stress tolerance during post-priming germination, a feature likely linked to a 'priming memory'.

  2. Accumulation of nitrogen - a critical parameter for the calculation of load limits from nitrogen in forests; Akkumulering av nitrogen - en kritisk parameter for beregning av taalegrenser for nitrogen i skog

    Sogn, T.A.; Stuanes, A.O.; Abrahamsen, G.


    The conference paper deals with the accumulation of nitrogen in forests in Norway. The level of accumulation is a critical factor for the calculation of load limits. The paper compares the average rapidity values of accumulation since the last glacial age with the calculated values from the more short-lasting period based on data from surveying programs of the State Pollution Control Authority, manuring experiments, and other relevant research programs in this field. 8 refs., 1 fig., 1 tab.

  3. The Ramachandran plots of glycine and pre-proline

    Brasseur Robert


    Full Text Available Abstract Background The Ramachandran plot is a fundamental tool in the analysis of protein structures. Of the 4 basic types of Ramachandran plots, the interactions that determine the generic and proline Ramachandran plots are well understood. The interactions of the glycine and pre-proline Ramachandran plots are not. Results In glycine, the ψ angle is typically clustered at ψ = 180° and ψ = 0°. We show that these clusters correspond to conformations where either the Ni+1 or O atom is sandwiched between the two Hα atoms of glycine. We show that the shape of the 5 distinct regions of density (the α, αL, βS, βP and βPR regions can be reproduced with electrostatic dipole-dipole interactions. In pre-proline, we analyse the origin of the ζ region of the Ramachandran plot, a region unique to pre-proline. We show that it is stabilized by a COi-1···CδHδi+1 weak hydrogen bond. This is analogous to the COi-1···NHi+1 hydrogen bond that stabilizes the γ region in the generic Ramachandran plot. Conclusion We have identified the specific interactions that affect the backbone of glycine and pre-proline. Knowledge of these interactions will improve current force-fields, and help understand structural motifs containing these residues.

  4. Effects of copper on chlorophyll, proline, protein and abscisic acid level of sunflower (Helianthus annuus L.) seedlings.

    Zengin, Fikriye Kirbag; Kirbag, Sevda


    The effect of copperchloride (CuCl2) on the level of chlorophyll (a+b), proline, protein and abscisic acid in sunflower (Helianthus annuus L.) seedlings were investigated Control and copper treated (0.4, 0.5 and 0.6 mM) seedlings were grown for ten days in Hoagland solution. Abscisic acid content was determined in root, shoot and leaf tissues of seedlings by HPLC. Copper stress caused significant increase of the abscisic acid contents in roots, shoots and leaves of seedlings. The increase was dependent on the copper salt concentration. Enhanced accumulation of proline in the leaves of seedlings exposed to copper was determined, as well as a decrease of chlorophyll (a+b) and total protein (p Helianthus annuus L.) seedlings. Thus, we assumed that copper levels increase above some critical points seedling growth get negative effects. This assumption is in line with previous findings.

  5. Development of a simple fluorescence-based microplate method for the high-throughput analysis of proline in wine samples.

    Robert-Peillard, Fabien; Boudenne, Jean-Luc; Coulomb, Bruno


    This paper presents a simple, accurate and multi-sample method for the determination of proline in wines thanks to a 96-well microplate technique. Proline is the most abundant amino acid in wine and is an important parameter related to wine characteristics or maturation processes of grape. In the current study, an improved application of the general method based on sodium hypochlorite oxidation and o-phthaldialdehyde (OPA)-thiol spectrofluorometric detection is described. The main interfering compounds for specific proline detection in wines are strongly reduced by selective reaction with OPA in a preliminary step under well-defined pH conditions. Application of the protocol after a 500-fold dilution of wine samples provides a working range between 0.02 and 2.90gL(-1), with a limit of detection of 7.50mgL(-1). Comparison and validation on real wine samples by ion-exchange chromatography prove that this procedure yields accurate results. Simplicity of the protocol used, with no need for centrifugation or filtration, organic solvents or high temperature enables its full implementation in plastic microplates and efficient application for routine analysis of proline in wines. Copyright © 2013 Elsevier Ltd. All rights reserved.

  6. Proline Sulfonamide-Based Organocatalysis: Better Late than Never.

    Yang, Hua; Carter, Rich G


    The history of proline sulfonamides for use in catalyzing highly enantioselective and diastereoselective C-C bond forming reactions is described. Highlighted is the development of N-(p-dodecylphenylsulfonyl)-2-pyrrolidinecarboxamide ("Hua Cat") and N-(carboxy-p-dodecylphenylsulfonyl)-2-pyrrolidinecarboxamide ("Hua Cat-II") by Yang and Carter. Specific reactions covered include the aldol reaction, Mannich reaction, formal aza-Diels-Alder reaction, tandem Michael/Mannich reaction and Yamada-Otani reaction. The ability of proline aryl sulfonamides to construct all-carbon quaternary stereocenters in high enantioselectivity and diastereoselectivity is a notable feature of the reported work. The practicality of this chemistry for large scale and industrial applications is also included. Finally, a discussion of the future directions of proline sulfonamide organocatalysis is provided.

  7. A theoretical study on interaction of proline with gold cluster

    Sandhya Rai; N V Suresh Kumar; Harjinder Singh


    Interaction of proline with gold cluster was studied using density functional theory (DFT). Two types of mixed basis sets UB3LYP/6-311++G ∪ LANL2MB and UB3LYP/6-311++G ∪ LANL2DZ were used for optimization of complex structures. Proline interacts with gold cluster either through one anchor bond, N–Au or an anchor bond O–Au associated with a non-conventional O–H…Au hydrogen bond. Among these interactions, higher tendency for interaction is seen with Au cluster through amide terminal. Natural bond orbital analysis (NBO) is used to substantiate the results.

  8. Effects of Chicken Manure and Mineral Fertilizer on Some Nutritive Parameters and Lead Accumulation in Two Vigna Species Grown in Lead Contaminated Soil

    N. Hamid


    Full Text Available Effect of chicken manure and mineral fertilizer on Vigna radiata and Vigna unguiculata plants grown in lead contaminated soil were examined. Plants were grown in lead polluted soil, which is amended with different concentrations of chicken manure and mineral fertilizer (NPK. Accumulation of lead in the leaves of both Vigna species was reduced when chicken manure was added. Application of mineral fertilizer increased the accumulation of lead in Vigna radiata, but caused a decrease in Vigna unguiculata. Application of both fertilizers improved the chlorophyll, protein and carbohydrate contents of both species. Results are discussed in relation to the significance of organic and inorganic manure with accumulation of heavy metals such as lead particularly in contaminated soils and their consequences on protein and chlorophyll synthesis.

  9. Nitrogen metabolism enzymes, soluble protein and free proline content in soybean genotypes and their F1 hybrids

    Kereši Sanja T.


    Full Text Available Nitrate reductase and glutamine synthetase activity, as well as free proline and soluble protein content were measured in eight soybean parent genotypes and six F1 hybrids. The aim of this study was to determine variability and the mode of inheritance for these parameters, and point out the genotypes of interest for future breeding programs. Analysed genotypes and their F1 hybrids expressed significant differences in activities of nitrate reductase and glutamine synthetase enzymes, as well as in soluble proteins and free proline contents. Since mode of inheritance for all investigated traits was in most cases dominance or heterosis, it can be concluded that these parameters are under control of dominant genes. The obtained results suggest that genotypes with favorable traits, such as variety Linda, line 1511, and F1 hybrids (Linda x LN92-7369 and (Balkan x BL-8, could be of interest as a good starting material for further breeding programs.

  10. Pyrroline-5-carboxylate synthase and proline biosynthesis: From osmotolerance to rare metabolic disease

    Pérez-Arellano, Isabel; Carmona-Álvarez, Francisco; Martínez, Ana I; Rodríguez-Díaz, Jesús; Cervera, Javier


    Pyrroline-5-carboxylate synthase (P5CS) is a bifunctional enzyme that exhibits glutamate kinase (GK) and γ-glutamyl phosphate reductase (GPR) activities. The enzyme is highly relevant in humans because it belongs to a combined route for the interconversion of glutamate, ornithine and proline. The deficiency of P5CS activity in humans is associated with a rare, inherited metabolic disease. It is well established that some bacteria and plants accumulate proline in response to osmotic stress. The alignment of P5CSs from different species and analysis of the solved structures of GK and GPR reveal high sequence and structural conservation. The information acquired from different mutant enzymes with increased osmotolerant properties, together with the position of the insertion found in the longer human isoform, permit the delimitation of the regulatory site of GK and P5CS and the proposal of a model of P5CS architecture. Additionally, the GK moiety of the human enzyme has been modeled and the known clinical mutations and polymorphisms have been mapped. PMID:20091669

  11. Molecular characterization of a proline transporter from Chilo suppressalis

    Jia Huang; Shun-Fan Wu; Gong-Yin Ye


    Proline plays an important role in regulating synaptic neurotransmission in both mammalian and insect nervous systems. The full-length complementary DNA sequences encoding the proline transporter gene (CsProT) and its alternative spliced transcript (CsProT-ST) were cloned from the striped stem borer (SSB), Chilo suppressalis,one of the most important rice pests in Asia. The comparison of amino acid sequences showed that CsProT is highly similar to the proline transporter isolated from Manduca sexta (MasProT). The developmental changes of the relative messenger RNA expression levels of CsProT and CsProT-ST genes were examined in SSB. The expression level of CsProT-ST is much higher than that of CsProT in all developmental periods, suggesting that the 3′-end spliced variant is the major transcript instead of CsProT. The highest expression level of both genes in the pupal stage means that proline may be involved in some unclear functions in the metamorphosis from pupa to adulthood.

  12. Screening and Characterization of Proline Dehydrogenase Flavoenzyme Producing Pseudomonas Entomophila

    H Shahbaz- Mohammadi


    Full Text Available Background and Objectives: Proline dehydrogenase (ProDH; plays an important role in specific determination of plasma proline level in biosensor and diagnostic kits. The goal of this research was to isolate and characterize ProDH enzyme from Iranian soil microorganisms.Materials and Methods: Screening of L-proline degradative enzymes from soil samples was carried out employing enrichment culture techniques. The isolate was characterized by biochemical reactions and specific PCR amplification. The target ProDH was purified and the effects of pH and temperature on the activity and stability were also tested.Results: Among the 250 isolates recovered from 40 soil samples, only one strain characterized as Pseudomonas entomophila displayed the highest enzyme activity toward L-proline (350 U/l than others. The enzyme of interest was identified as a ProDH and had Km value of 32 mM for L-proline. ProDH exhibited its best activity at temperature range of 25 to 35°C and its highest activity was achieved at 30°C. It was almost stable at temperatures between 25-30°C for 2 hours. The optimum pH activity of ProDH reaction was 8.5 and its activity was stable in pH range of 8.0-9.0 upto 24 hours. The enzyme was purified with a yield of 8.5% and a purification factor of 37.7. The molecular mass of the purified ProDH was about 40 kDa, and determined to be a monomeric protein."nConclusion: This is the first report concerning the ProDH production by a P. entomophila bacterium isolated from soil sample.

  13. Molybdenum (Mo) increases endogenous phenolics, proline and photosynthetic pigments and the phytoremediation potential of the industrially important plant Ricinus communis L. for removal of cadmium from contaminated soil.

    Hadi, Fazal; Ali, Nasir; Fuller, Michael Paul


    Cadmium (Cd) in agricultural soil negatively affects crops yield and compromises food safety. Remediation of polluted soil is necessary for the re-establishment of sustainable agriculture and to prevent hazards to human health and environmental pollution. Phytoremediation is a promising technology for decontamination of polluted soil. The present study investigated the effect of molybdenum (Mo) (0.5, 1.0 and 2.0 ppm) on endogenous production of total phenolics and free proline, plant biomass and photosynthetic pigments in Ricinus communis plants grown in Cd (25, 50 and 100 ppm) contaminated soils and the potential for Cd phytoextraction. Mo was applied via seed soaking, soil addition and foliar spray. Foliar sprays significantly increased plant biomass, Cd accumulation and bioconcentration. Phenolic concentrations showed significantly positive correlations with Cd accumulation in roots (R (2) = 0.793, 0.807 and 0.739) and leaves (R (2) = 0.707, 721 and 0.866). Similarly, proline was significantly positively correlated with Cd accumulation in roots (R (2) = 0.668, 0.694 and 0.673) and leaves (R (2) = 0.831, 0.964 and 0.930). Foliar application was found to be the most effective way to deliver Mo in terms of increase in plant growth, Cd accumulation and production of phenolics and proline.

  14. Proline derivatives in fruits of bergamot (Citrus bergamia Risso et Poit): presence of N-methyl-L-proline and 4-hydroxy-L-prolinebetaine.

    Servillo, Luigi; Giovane, Alfonso; Balestrieri, Maria Luisa; Cautela, Domenico; Castaldo, Domenico


    The content of proline and various compounds deriving from its metabolism (4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine) was determined in fruits and seeds of Bergamot (Citrus bergamia Risso et Poit), growing in the Calabria region (South Italy). A HPLC-ESI-tandem mass spectrometry method, which allowed rapid determination of L-proline, 4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine in juice and extracts of bergamot fruit with minimum sample preparation and short analysis time (about 10 min), is presented. Proline and 4-hydroxy-L-proline levels in the samples were also determined by HPLC analysis with fluorescence detection and the results compared to those obtained with HPLC-ESI-tandem mass spectrometry. For the first time, the presence of N-methyl-L-proline and 4-hydroxy-L-prolinebetaine in the fruits of a plant of the Citrus genus is reported.

  15. Modulation of physicochemical and spectroscopic properties of l-serine and l-proline by propionate based food preservatives.

    Banipal, Tarlok S; Kaur, Navalpreet; Kaur, Jaspreet; Komal; Banipal, Parampaul K


    To have an insight into the effect of preservatives on various ingredients of processed items, it is important to study their thermodynamic, transport and spectroscopic properties in aqueous solutions to elucidate various solute-co-solute interactions. The densities, viscosities and enthalpies of dilution of l-serine and l-proline have been determined in water and in aqueous solutions of sodium propionate and calcium propionate at different temperatures. The derived parameters elucidate the changes in taste quality and hydration number of l-serine and l-proline in the presence of the studied preservatives. Predominance of dehydration effect has been observed from calorimetry and changes in chemical shifts from nuclear magnetic resonance spectroscopy also support the above results.

  16. [The main pharmacokinetic parameters of p-tyrosol upon intravenous injection in rats. II. Verification of the pharmacokinetics linearity and evaluation of the possible accumulation].

    Chernysheva, G A; Plotnikov, M B; Smol'iakova, V I; Cherkashina, I V; Tolstikova, T G; Krysin, A P; Sorokina, I V


    The main pharmacokinetic parameters of p-tyrosol after single (in 3 doses) and repeated intravenous injection were studied in rats. The content ofp-tyrosol in the blood plasma was determined by spectrofluorimetric method. The pharmacokinetic parameters of p-tyrosol are linear in the dose range from 50 to 200 mg/kg. Repeated administration leads to accelerated metabolic elimination of p-tyrosol.

  17. The role of proline in the prevention of aggregation during protein folding in vitro.

    Kumar, T K; Samuel, D; Jayaraman, G; Srimathi, T; Yu, C


    Proline effectively inhibits protein aggregation during the refolding of bovine carbonic anhydrase. Other osmolytes used such as glycine and ethylene glycol fail to exhibit the 'aggregation-blockade' role shown by proline. Results of viscosity and ANS fluorescence (1-anilino-8-naphthalene sulphonic acid) experiments suggest that proline at high concentrations forms an ordered supramolecular assembly. Based on these results, it is proposed that proline behaves as a protein folding chaperone due to the formation of an ordered, amphipathic supramolecular assembly. To our knowledge, this is the first report wherein proline is proposed as a protein folding aid.

  18. On the temporal variation of leaf magnetic parameters: seasonal accumulation of leaf-deposited and leaf-encapsulated particles of a roadside tree crown.

    Hofman, Jelle; Wuyts, Karen; Van Wittenberghe, Shari; Samson, Roeland


    Understanding the accumulation behaviour of atmospheric particles inside tree leaves is of great importance for the interpretation of biomagnetic monitoring results. In this study, we evaluated the temporal variation of the saturation isothermal remanent magnetisation (SIRM) of leaves of a roadside urban Platanus × acerifolia Willd. tree in Antwerp, Belgium. We hereby examined the seasonal development of the total leaf SIRM signal as well as the leaf-encapsulated fraction of the deposited dust, by washing the leaves before biomagnetic analysis. On average 38% of the leaf SIRM signal was exhibited by the leaf-encapsulated particles. Significant correlations were found between the SIRM and the cumulative daily average atmospheric PM10 and PM2.5 measurements. Moreover, a steady increase of the SIRM throughout the in-leaf season was observed endorsing the applicability of biomagnetic monitoring as a proxy for the time-integrated PM exposure of urban tree leaves. Strongest correlations were obtained for the SIRM of the leaf-encapsulated particles which confirms the dynamic nature of the leaf surface-accumulated particles.

  19. Cyclophilin A catalyzes proline isomerization by an electrostatic handle mechanism

    Camilloni, Carlo; Sahakyan, Aleksander B.; Holliday, Michael; Isern, Nancy G.; Zhang, Fengli; Eisenmesser, Elan Z.; Vendruscolo, Michele


    Proline isomerization is a ubiquitous process that plays a key role in the folding of proteins and in the regulation of their functions1-3. Different families of enzymes, known as peptidyl-prolyl isomerases (PPIases), catalyse this reaction, which involves the interconversion between the cis and trans isomers of the Nterminal amide bond of the amino acid proline2,3. A complete description of the mechanisms by which these enzymes function, however, has remained elusive. Here, we show that cyclophilin A, one of the most common PPIases4, provides a catalytic environment that acts on the substrate through an electrostatic lever mechanism. In this mechanism, the electrostatic field in the catalytic site turns the electric dipole associated with the carboxylic group of the amino acid preceding the proline in the substrate, thus causing the rotation of the peptide bond between the two residues. This mechanism resulted from the analysis of an ensemble of conformations populated by cyclophilin A during the enzymatic reaction using a combination of NMR measurements, molecular dynamics simulations and density functional theory calculations. We anticipate that this approach will be helpful in elucidating whether the electrostatic lever mechanism that we describe is common to other PPIases, and more generally to characterise other enzymatic processes.

  20. A case study of proline isomerization in cell signaling.

    Min, Lie; Fulton, D Bruce; Andreotti, Amy H


    Protein-mediated interactions and enzymatic function provide the foundation upon which cellular signaling cascades control all of the activities of a cell. Post-translational modifications such as phosphorylation or ubiquitiation are well known means for modulating protein activity within the cell. These chemical modifications create new recognition motifs on proteins or shift conformational preferences such that protein catalytic and binding functions are altered in response to external stimuli. Moreover, detection of such modifications is often straightforward by conventional biochemical methods leading investigators toward mechanistic models of cell signaling involving post-translational modifications such as phosphorylation/dephosphorylation. While there is little doubt that such modifications play a significant role in transmission of information throughout the cell, there are certainly other mechanisms at work that are not as well understood at this time. Of particular interest in the context of this review is the intrinsic conformational switch afforded to a polypeptide by peptidyl prolyl cis/trans isomerization. Proline isomerization is emerging as a critical component of certain cell signaling cascades. In addition to serving as a conformational switch that enables a protein to adopt functionally distinct states, proline isomerization may serve as a recognition element for the ubiquitous peptidyl prolyl isomerases. This overview takes a close look at one particular signaling protein, the T cell specific tyrosine kinase Itk, and examines the role of proline isomerization and the peptidyl prolyl isomerase cyclophilin A in mediating Itk function following T cell receptor engagement.

  1. The Combined Effects of Arbuscular Mycorrhizal Fungi (AMF and Lead (Pb Stress on Pb Accumulation, Plant Growth Parameters, Photosynthesis, and Antioxidant Enzymes in Robinia pseudoacacia L.

    Yurong Yang

    Full Text Available Arbuscular mycorrhizal fungi (AMF are considered as a potential biotechnological tool for improving phytostabilization efficiency and plant tolerance to heavy metal-contaminated soils. However, the mechanisms through which AMF help to alleviate metal toxicity in plants are still poorly understood. A greenhouse experiment was conducted to evaluate the effects of two AMF species (Funneliformis mosseae and Rhizophagus intraradices on the growth, Pb accumulation, photosynthesis and antioxidant enzyme activities of a leguminous tree (Robinia pseudoacacia L. at Pb addition levels of 0, 500, 1000 and 2000 mg kg(-1 soil. AMF symbiosis decreased Pb concentrations in the leaves and promoted the accumulation of biomass as well as photosynthetic pigment contents. Mycorrhizal plants had higher gas exchange capacity, non-photochemistry efficiency, and photochemistry efficiency compared with non-mycorrhizal plants. The enzymatic activities of superoxide dismutase (SOD, ascorbate peroxidases (APX and glutathione peroxidase (GPX were enhanced, and hydrogen peroxide (H2O2 and malondialdehyde (MDA contents were reduced in mycorrhizal plants. These findings suggested that AMF symbiosis could protect plants by alleviating cellular oxidative damage in response to Pb stress. Furthermore, mycorrhizal dependency on plants increased with increasing Pb stress levels, indicating that AMF inoculation likely played a more important role in plant Pb tolerance in heavily contaminated soils. Overall, both F. mosseae and R. intraradices were able to maintain efficient symbiosis with R. pseudoacacia in Pb polluted soils. AMF symbiosis can improve photosynthesis and reactive oxygen species (ROS scavenging capabilities and decrease Pb concentrations in leaves to alleviate Pb toxicity in R. pseudoacacia. Our results suggest that the application of the two AMF species associated with R. pseudoacacia could be a promising strategy for enhancing the phytostabilization efficiency of Pb

  2. Accumulator Parameters Match the Characteristic Analysis of Hydraulic Brake for Trackless Rubber Tire Vehicle%全液压制动无轨胶轮车载蓄能器参数匹配与特性分析



    根据目前无轨胶轮车全液压制动系统的使用情况,建立了蓄能器的数学模型,通过分析液压蓄能器参数匹配和能量利用对制动系统动态性能的影响,为液压制动系统蓄能器设计提供理论依据.%According to the current use of trackless rubber tire vehide hydraulic braking system, the mathematical model of the accumulator, hydraulic accumulator parameters by analyzing the matchand use of energy on the dynamic performance of the brake system, hydraulic brake system reservoir design can provide a theoretical basis.

  3. Ectoine accumulation in Brevibacterium epidermis.

    Onraedt, Annelies; De Muynck, Cassandra; Walcarius, Bart; Soetaert, Wim; Vandamme, Erick


    As a halotolerant bacterial species, Brevibacterium epidermis DSM 20659 can grow at relatively high salinity, tolerating up to 2 M NaCl. It synthesizes ectoine and the intracellular content increases with the medium salinity, with a maximum of 0.14 g ectoine/g CDW at 1 M NaCl. Sugar-stressed cells do not synthesize ectoine. Ectoine synthesis is also affected by the presence of external osmolytes. Added betaine is taken up and completely replaced ectoine, while L-proline is only temporarily accumulated after which ectoine is synthesized. The strain can metabolize ectoine; L-glutamate is a better carbon source for ectoine synthesis than L-aspartate.

  4. Evidence that COMT genotype and proline interact on negative-symptom outcomes in schizophrenia and bipolar disorder

    Clelland, C L; Drouet, V.; Rilett, K C; Smeed, J A; Nadrich, R H; Rajparia, A; Read, L L; Clelland, J D


    Elevated peripheral proline is associated with psychiatric disorders, and there is evidence that proline is a neuromodulator. The proline dehydrogenase (PRODH) gene, which encodes the enzyme that catalyzes proline catabolism, maps to human chromosome 22q11.2, a region conferring risk of schizophrenia. In the Prodh-null mouse, an interaction between elevated peripheral proline and another 22q11.2 gene, catechol-O-methyltransferase (COMT), on neurotransmission and behavior has been reported. We...

  5. Accumulation of nickel ions in seedlings of Vicia sativa L. and manifestations of oxidative stress.

    Ivanishchev, V V; Abramova, E A


    The accumulation of nickel ions in the roots and shoots of vetch seedlings (Vicia sativa L.) at increasing concentrations of nickel chloride in the medium was studied. It was shown that the accumulation of nickel in the shoots was increased when the concentration of nickel chloride in the medium was more than 50 μM. The bioconcentration factor and sustainability index for vetch seedlings were calculated under the experimental conditions. The obtained results were similar to parameters for other plants, grown on a nutrient medium or soil substrate. First, the obtained results allowed estimate the limits of nickel chloride concentrations for four of five zones, which correspond to the theoretical concept of dose-response curves in the studies on the influence of physiologically essential heavy metals on plants (Prasad 2010). Some parameters of oxidative stress caused by the presence of nickel chloride in the medium were shown. It seems that at low nickel concentrations in the medium in vetch seedlings the increase of several biochemical parameters (catalase activity and proline) caused by the high amylase activity in seeds.


    Iwona Kozikowska


    Full Text Available The aim of this study was to determine the content of magnesium, copper, cadmium and iron in umbilical cord blood of the newborns depending on the number of pregnancies. It was established correlations between average concentrations these metals in cord blood and newborn’s parameters. The study material was collected immediately after delivery from the Department of Obstetrics and Gynecology in Bytom. The cord blood was taken from 99 women between 29-40 years old. Women were divided into two groups: nulliparous and multiparous. The concentration of metals in the cord blood was determined by flame atomic absorption spectrometry (FAAS. The conducted study demonstrates that magnesium, copper, cadmium and iron were noted in all tissues, both nulliparous women and multiparous mothers. The maximum concentration of cadmium in umbilical cord blood was observed among multiparous mothers (2.229 d.m.. In group of nulliparous women was observed higher concentration of Fe, Mg and Cu in umbilical cord blood than in multiparous mothers. It was noted some statistically significant correlation between iron, copper and newborn’s parameters. Parity influences the concentration of cadmium in umbilical cord blood with higher level found in multiparous women. Average content of iron in cord blood did not decrease with parity, it indicate that this element is preferentially taken up by the child.

  7. Supramolecular properties of the proline-rich gamma-Zein N-terminal domain.

    Kogan, Marcelo J; Dalcol, Ionara; Gorostiza, Pau; Lopez-Iglesias, Carmen; Pons, Ramon; Pons, Miquel; Sanz, Fausto; Giralt, Ernest


    Zeins are maize storages proteins that accumulate inside large vesicles called protein bodies. gamma-Zein lines the inner face of the protein body membrane, and its N-terminal proline-rich repetitive domain with the sequence (VHLPPP)(8) appears to be necessary for the accumulation of the protein within the organelle. Synthetic (VHLPPP)(8) adopts an amphipathic polyproline II conformation. In a preliminary recent work we used atomic force microscopy to study the surface organization of the octamer and transmission electron microscopy to visualize aggregates of the peptide from aqueous solution. We previously envisioned two self-assembly models (i.e., the geometric and the micellar) that take into account the observed features. In the present work we studied in detail the self-assembly of the peptide in solution and found that the peptide is able to form cylindrical micelles. Fibrils formed on graphite are generated by assembly of solution micelles. Based on the results of these studies, we focused exclusively on the micellar model. To our knowledge we have characterized for the first time supramolecular aggregates of polyproline structures other than collagen. The spontaneous arrangement of (VHLPPP)(8) suggests a role for the N-terminal domain of gamma-zein in the process of the whole protein deposition in protein bodies.

  8. Reduced collagen accumulation after major surgery

    Jorgensen, L N; Kallehave, F; Karlsmark, T


    The preoperative and postoperative wound-healing capacity of 23 patients undergoing elective major abdominal, thoracic or urological surgery was tested objectively by the subcutaneous accumulation of hydroxyproline and proline in an expanded polytetrafluoroethylene (ePTFE) tube. Before scheduled...... surgery two ePTFE tubes were implanted for removal after 5 and 10 days. This was repeated for each patient immediately after surgery. After 10 days a higher amount of hydroxyproline was measured before than after operation (median 2.91 (range 0.37-14.45) versus 1.45 (range 0.26-6.94) micrograms/cm, P = 0...

  9. Exposure of Brassica juncea (L) to arsenic species in hydroponic medium: comparative analysis in accumulation and biochemical and transcriptional alterations.

    Ahmad, Mohd Anwar; Gupta, Meetu


    Arsenic (As) contamination in the environment has attracted considerable attention worldwide. The objective of the present study was to see the comparative effect of As species As(III) and As(V) on accumulation, biochemical responses, and gene expression analysis in Brassica juncea var. Pusa Jaganath (PJn). Hydroponically grown 14-day-old seedlings of B. juncea were treated with different concentrations of As(III) and As(V). Accumulation of total As increased with increasing concentration of both As species and exposure time, mainly in roots. Reduction in seed germination, root-shoot length, chlorophyll, and protein content were observed with increasing concentration and exposure time of both As species, being more in As(III)-treated leaves. PJn variety showed that antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) and stress-related parameters (cysteine, proline, and malondialdehyde (MDA)) were stimulated and allows plant to tolerate both As species. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis in leaves showed significant changes in protein profile with more stringent effect with As(III) stress. Semiquantitative RT-PCR analysis showed regulation in expression of phytochelatin synthase (PCS), metallothionine-2 (MT-2), glutathione reductase (GR), and glutathione synthetase (GS) genes under both As(III) and As(V) stresses. Results suggested that accumulation and inhibition on physiological parameters differ according to the As species, while molecular and biochemical parameters showed a combinatorial type of tolerance mechanism against As(III) and As(V) stresses.

  10. Influence of the Lactotripeptides Isoleucine-Proline-Proline and Valine-Proline-Proline on Systolic Blood Pressure in Japanese Subjects: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

    Aurelie Chanson-Rolle

    Full Text Available The lactotripeptides isoleucine-proline-proline (IPP and valine-proline-proline (VPP have been shown to decrease systolic blood pressure (SBP in several populations, but the size of the effect varies among studies. We performed a meta-analysis including all published studies to evaluate the SBP-lowering effect of IPP/VPP in Japanese subjects more comprehensively.Eligible randomized controlled trials were searched for within four bibliographic databases, including two Japanese ones. Eighteen studies (including a total of 1194 subjects were included in the meta-analysis. A random effect model using the restricted maximum likelihood (REML estimator was used for the analysis. The analysis showed that consumption of IPP/VPP induced a significant reduction in SBP as compared with placebo in Japanese subjects, with an estimated effect of -5.63 mm Hg (95% CI, -6.87 to -4.39, P<0.0001 and no evidence of publication bias. A significant heterogeneity between series was evident, which could be explained by a significant influence of the baseline blood pressure status of the subjects, the effect of IPP/VPP on SBP being stronger in hypertensive subjects (-8.35 mm Hg, P<0.0001 than in non-hypertensive subjects (-3.42mm Hg, P<0.0001. Furthermore, the effect of IPP/VPP on SBP remained significant when limiting the analysis to series that tested the usual doses of IPP/VPP consumed daily (below 5 mg/d, with estimated effects of -6.01 mm Hg in the overall population and -3.32 mm Hg in non-hypertensive subjects.Results from this meta-analysis show that IPP/VPP lactotripeptides can significantly reduce office SBP in Japanese subjects with or without overt hypertension, and for doses that can potentially be consumed as an everyday supplement. This suggests that these peptides could play a role in controlling blood pressure in Japanese subjects. The systematic review protocol was published on the PROSPERO register (CRD42014014322.

  11. Sources of superoxide/H2O2 during mitochondrial proline oxidation

    Renata L.S. Goncalves


    Full Text Available p53 Inducible gene 6 (PIG6 encodes mitochondrial proline dehydrogenase (PRODH and is up-regulated several fold upon p53 activation. Proline dehydrogenase is proposed to generate radicals that contribute to cancer cell apoptosis. However, there are at least 10 mitochondrial sites that can produce superoxide and/or H2O2, and it is unclear whether proline dehydrogenase generates these species directly, or instead drives production by other sites. Amongst six cancer cell lines, ZR75-30 human breast cancer cells had the highest basal proline dehydrogenase levels, and mitochondria isolated from ZR75-30 cells consumed oxygen and produced H2O2 with proline as sole substrate. Insects use proline oxidation to fuel flight, and mitochondria isolated from Drosophila melanogaster were even more active with proline as sole substrate than ZR75-30 mitochondria. Using mitochondria from these two models we identified the sites involved in formation of superoxide/H2O2 during proline oxidation. In mitochondria from Drosophila the main sites were respiratory complexes I and II. In mitochondria from ZR75-30 breast cancer cells the main sites were complex I and the oxoglutarate dehydrogenase complex. Even with combinations of substrates and respiratory chain inhibitors designed to minimize the contributions of other sites and maximize any superoxide/H2O2 production from proline dehydrogenase itself, there was no significant direct contribution of proline dehydrogenase to the observed H2O2 production. Thus proline oxidation by proline dehydrogenase drives superoxide/H2O2 production, but it does so mainly or exclusively by providing anaplerotic carbon for other mitochondrial dehydrogenases and not by producing superoxide/H2O2 directly.

  12. Sources of superoxide/H2O2 during mitochondrial proline oxidation.

    Goncalves, Renata L S; Rothschild, Daniel E; Quinlan, Casey L; Scott, Gary K; Benz, Christopher C; Brand, Martin D


    p53 Inducible gene 6 (PIG6) encodes mitochondrial proline dehydrogenase (PRODH) and is up-regulated several fold upon p53 activation. Proline dehydrogenase is proposed to generate radicals that contribute to cancer cell apoptosis. However, there are at least 10 mitochondrial sites that can produce superoxide and/or H2O2, and it is unclear whether proline dehydrogenase generates these species directly, or instead drives production by other sites. Amongst six cancer cell lines, ZR75-30 human breast cancer cells had the highest basal proline dehydrogenase levels, and mitochondria isolated from ZR75-30 cells consumed oxygen and produced H2O2 with proline as sole substrate. Insects use proline oxidation to fuel flight, and mitochondria isolated from Drosophila melanogaster were even more active with proline as sole substrate than ZR75-30 mitochondria. Using mitochondria from these two models we identified the sites involved in formation of superoxide/H2O2 during proline oxidation. In mitochondria from Drosophila the main sites were respiratory complexes I and II. In mitochondria from ZR75-30 breast cancer cells the main sites were complex I and the oxoglutarate dehydrogenase complex. Even with combinations of substrates and respiratory chain inhibitors designed to minimize the contributions of other sites and maximize any superoxide/H2O2 production from proline dehydrogenase itself, there was no significant direct contribution of proline dehydrogenase to the observed H2O2 production. Thus proline oxidation by proline dehydrogenase drives superoxide/H2O2 production, but it does so mainly or exclusively by providing anaplerotic carbon for other mitochondrial dehydrogenases and not by producing superoxide/H2O2 directly.

  13. Prediction and Biochemical Demonstration of a Catabolic Pathway for the Osmoprotectant Proline Betaine

    Kumar, Ritesh; Zhao, Suwen; Vetting, Matthew W.; Wood, B. McKay; Sakai, Ayano; Cho, Kyuil; Solbiati, José; Steven C Almo; Jonathan V Sweedler; Matthew P Jacobson; Gerlt, John A.; Cronan, John E.


    ABSTRACT Through the use of genetic, enzymatic, metabolomic, and structural analyses, we have discovered the catabolic pathway for proline betaine, an osmoprotectant, in Paracoccus denitrificans and Rhodobacter sphaeroides. Genetic and enzymatic analyses showed that several of the key enzymes of the hydroxyproline betaine degradation pathway also function in proline betaine degradation. Metabolomic analyses detected each of the metabolic intermediates of the pathway. The proline betaine catab...

  14. Oxygen Reactivity of PutA from Helicobacter Species and Proline-Linked Oxidative Stress

    Krishnan, Navasona; Becker, Donald F.


    Proline is converted to glutamate in two successive steps by the proline utilization A (PutA) flavoenzyme in gram-negative bacteria. PutA contains a proline dehydrogenase domain that catalyzes the flavin adenine dinucleotide (FAD)-dependent oxidation of proline to Δ1-pyrroline-5-carboxylate (P5C) and a P5C dehydrogenase domain that catalyzes the NAD+-dependent oxidation of P5C to glutamate. Here, we characterize PutA from Helicobacter hepaticus (PutAHh) and Helicobacter pylori (PutAHp) to pro...

  15. Thermoresponsive and Mechanical Properties of Poly(L-proline) Gels.

    Gkikas, Manos; Avery, Reginald K; Olsen, Bradley D


    Gelation of the left helical N-substituted homopolypeptide poly(L-proline) (PLP) in water was explored, employing rheological and small-angle scattering studies at different temperatures and concentrations in order to investigate the network structure and its mechanical properties. Stiff gels were obtained at 10 wt % or higher at 5 °C, the first time gelation has been observed for homopolypeptides. The secondary structure and helical rigidity of PLP has large structural similarities to gelatin but as gels the two materials show contrasting trends with temperature. With increasing temperature in D2O, the network stiffens, with broad scattering features of similar correlation length for all concentrations and molar masses of PLP. A thermoresponsive transition was also achieved between 5 and 35 °C, with moduli at 35 °C higher than gelatin at 5 °C. The brittle gels could tolerate strains of 1% before yielding with a frequency-independent modulus over the observed range, similar to natural proline-rich proteins, suggesting the potential for thermoresponsive or biomaterial-based applications.

  16. Effects of Salt and Water Stress on Plant Growth and on Accumulation of Osmolytes and Antioxidant Compounds in Cherry Tomato

    Mohamad AL HASSAN


    Full Text Available The effects of salt and water stress on growth and several stress markers were investigated in cherry tomato plants. Some growth parameters (stem length and number of leaves and chlorophyll contents were determined every third day during plant growth, and leaf material was collected after 25 and 33 days of treatment. Both stresses inhibited plant growth; chlorophyll levels, however, decreased only in response to high NaCl concentrations. Proline contents largely increased in leaves of stressed plants, reaching levels high enough to play a major role in cellular osmotic adjustment. Despite reports indicating that tomato does not synthesize glycine betaine, the stress-induced accumulation of this osmolyte was detected in cherry tomato, albeit at lower concentration than that of proline. Therefore, it appears that the plants are able to synthesise glycine betaine as a secondary osmolyte under strong stress conditions. Total sugars levels, on the contrary, decreased in stress-treated plants. Both stress treatments caused secondary oxidative stress in the plants, as indicated by a significant increase in malondialdehyde (MDA contents. Water stress led to an increase in total phenolics and flavonoid contents and a reduction of carotenoid levels in the leaves; flavonoids also increased under high salinity conditions.

  17. Chronic Exposure to Proline Causes Aminoacidotoxicity and Impaired Beta-Cell Function: Studies In Vitro

    Liu, Zhenping; Jeppesen, Per Bendix; Gregersen, Søren;


    thymidine incorporation, and (5) protein expression of INS-1E cells in response to proline by proteomics. RESULTS: We found that high doses of proline increased BIS and decreased GSIS in both isolated mouse islets and INS-1E cells. MafA, insulin 1, and the cytochrome c oxidase subunit VIa polypeptide 2 m...

  18. Aqueous solutions of proline and NaCl studied by differential scanning calorimetry at subzero temperatures

    Rasmussen, Peter Have; Jørgensen, Bo; Nielsen, Jette


    The hydration properties of proline are studied by differential scanning calorimetry (DSC) in aqueous solutions during freezing to -60 degrees C and subsequent heating to +20 degrees C. The concentration of proline in the freeze concentrated solution was estimated to approximately 50 wt% (w/w) in...

  19. Cotton Effect in Copper-Proline Complexes in the Visible Region

    Volkov, Victor; Pfister, Rolf


    The electronic properties of Cu(II) complex with proline are considered to demonstrate the Cotton effect in the visible region. A series of experiments in optical rotatory dispersion spectroscopy with free D- and L-proline and their complexes with the Cu(II) ion in aqueous solution is suggested.

  20. Localization of two post-proline cleaving peptidases in the midgut of Tenebrio molitor larvae

    Two soluble post-proline cleaving peptidase activities, PPCP1 and PPCP2, were demonstrated in the midgut of Tenebrio molitor larvae with the substrate benzyloxycarbonyl-L-alanyl-L-proline p-nitroanilide. Both activities were serine peptidases. PPCP1 was active in acidic buffers, with maximum activit...

  1. Evaluation of potential cationic probes for the detection of proline and betaine.

    Kalsoom, Umme; Breadmore, Michael C; Guijt, Rosanne M; Boyce, Mary C


    Osmoregulants are the substances that help plants to tolerate environmental extremes such as salinity and drought. Proline and betaine are two of the most commonly studied osmoregulants. An indirect UV CE method has been developed for simultaneous determination of these osmoregulants. A variety of reported probes and compounds were examined as potential probes for the indirect detection of proline and betaine. Mobility and UV-absorption properties highlighted sulfanilamide as a potential probe for indirect analysis of proline and betaine. Using 5 mM sulfanilamide at pH 2.2 with UV detection at 254 nm, proline and betaine were separated in less than 15 min. The LODs for proline and betaine were 11.6 and 28.3 μM, respectively. The developed method was successfully applied to quantification of these two osmoregulants in spinach and beetroot samples.

  2. Proline Based Chiral Ionic Liquids for Enantioselective Michael Reaction

    Kaoru Nobuoka


    Full Text Available Chiral ionic liquids, starting from (S-proline, have been prepared and evaluated the ability of a chiral catalyst. In Michael reaction of trans-β-nitrostyrene and cyclohexanone, all the reactions were carried out under homogeneous conditions without any solvent except for excess cyclohexanone. The chiral ionic liquid catalyst with the positive charge delocalized bulky pyrrolidinium cation shows excellent yields (up to 92%, diastereoselectivities (syn/anti = 96/4, and enantioselectivities (up to 95% ee and could be reused at least three times without any loss of its catalytic activity. Such results demonstrated a promising new approach for green and economic chiral synthesis by using the chiral ionic liquids as a chiral catalyst and a chiral medium.

  3. Proline-catalysed asymmetric ketol cyclizations: The template mechanism revisited

    R Malathi; D Rajagopal; Zoltan G Hajos; S Swaminathan


    A modified template mechanism based on modelling studies of energy minimised complexes is presented for the asymmetric proline-catalysed cyclization of triketones 1, 2 and 3 to the 2, 3-ketols 1a, 2a and 3a respectively. The template model involves a three-point contact as favoured in enzyme-substrate interactions. Our minimisation studies are in agreement with the divergent behaviour of the 6,5-, 6,6- and 6,7-bicyclic systems. They support the high 93.4% ee observed with the 6,5-bicyclic ketol and the lower 73% ee found with the 6,6-bicyclic ketol. The calculations also explain the lack of asymmetric induction with the 6,7-bicyclic system.

  4. Oxidized low-density lipoproteins upregulate proline oxidase to initiate ROS-dependent autophagy.

    Zabirnyk, Olga; Liu, Wei; Khalil, Shadi; Sharma, Anit; Phang, James M


    Epidemiological studies showed that high levels of oxidized low-density lipoproteins (oxLDLs) are associated with increased cancer risk. We examined the direct effect of physiologic concentrations oxLDL on cancer cells. OxLDLs were cytotoxic and activate both apoptosis and autophagy. OxLDLs have ligands for peroxisome proliferator-activated receptor gamma and upregulated proline oxidase (POX) through this nuclear receptor. We identified 7-ketocholesterol (7KC) as a main component responsible for the latter. To elucidate the role of POX in oxLDL-mediated cytotoxicity, we knocked down POX via small interfering RNA and found that this (i) further reduced viability of cancer cells treated with oxLDL; (ii) decreased oxLDL-associated reactive oxygen species generation; (iii) decreased autophagy measured via beclin-1 protein level and light-chain 3 protein (LC3)-I into LC3-II conversion. Using POX-expressing cell model, we established that single POX overexpression was sufficient to activate autophagy. Thus, it led to autophagosomes accumulation and increased conversion of LC3-I into LC3-II. Moreover, beclin-1 gene expression was directly dependent on POX catalytic activity, namely the generation of POX-dependent superoxide. We conclude that POX is critical in the cellular response to the noxious effects of oxLDL by activating protective autophagy.

  5. The effect of methyl jasmonate and different chloride concentrations on photosynthetic pigments and proline content in Nicotiana tabacum L. cv. Cooker 347

    Mahyar Moshtaghi


    Full Text Available Jasmonic acid and its methylated ester (methyl jasmonate, as natural plant growth regulators, are widely found in plants. In this study, the effect of methyl jasmonate (30 μM, in the early stages of Nicotiana tabacum L. cv. Cooker 347 growth, on photosynthetic pigments and proline was studied at high concentration of chloride. This experiment was performed in randomized complete block design with 4 replications, during 2011 crop year at Guilan Tobacco Research Center, outside greenhouse and under pot conditions. After transplanting similar seedlings to pots in rapid growth phase, the plants were irrigated with three levels chloride (50, 150 and 300 mg/L as CaCl2 both in presence and absence of methyl jasmonate for 4 weeks. Data analysis showed that by increasing Cl- concentrations up to 300 (mg/L, chloride and calcium accumulation of leaves increased, whilst photosynthetic pigments and proline decreased. Exogenous application of methyl jasmonate (30 μM significantly increased photosynthetic pigments in all treatments and proline in the 300 mg/L treatment. It seemed that the application of external methyl jasmonate (30 µM could improve tobacco plant injuries at high chloride concentration.

  6. Pre-storage application of oxalic acid alleviates chilling injury in mango fruit by modulating proline metabolism and energy status under chilling stress.

    Li, Peiyan; Zheng, Xiaolin; Liu, Yan; Zhu, Yuyan


    Effects of oxalic acid on chilling injury, proline metabolism and energy status in mango fruit were investigated after mango fruit (Mangifera indica L. cv. Zill) were dipped in 5mM oxalic acid solution for 10min at 25°C and then stored at low temperature (10±0.5°C) for 49days thereafter transferred to 25°C for 4days. Pre-storage application of oxalic acid apparently inhibited the development of chilling injury, notably elevated proline accumulation actually associated with increase in Δ(1)-pyrroline-5-carboxylate synthetase (P5CS) activity and decrease in proline dehydrogenase (PDH) activity in the peel and the flesh, without activation of ornithine-δ-aminotransferase (OAT) activity, and maintained high ATP level and energy charge in the flesh during storage. It was suggested that these effects of oxalic acid might collectively contribute to improving chilling tolerance, thereby alleviating chilling injury and maintaining quality of mango fruit in long term cold storage.

  7. Effect of ultraviolet radiation on chlorophyll, carotenoid, protein and proline contents of some annual desert plants.

    Salama, Hediat M H; Al Watban, Ahlam A; Al-Fughom, Anoud T


    Investigation was carried out to find whether enhanced ultraviolet radiation influences the Malva parviflora L., Plantago major L., Rumex vesicarius L. and Sismbrium erysimoids Desf. of some annual desert plants. The seeds were grown in plastic pots equally filled with a pre-sieved normal sandy soil for 1 month. The planted pots from each species were randomly divided into equal groups (three groups). Plants of the first group exposed to white-light tubes (400-700 nm) 60 w and UV (365 nm) 8 w tubes. The second group was exposed to white-light tubes (400-700 nm) 60 w and UV (302 nm) 8 w tubes. The third group was exposed to white-light tubes (400-700 nm) 60 w and UV (254 nm) 8 w tubes, respectively, for six days. The results indicated that the chlorophyll contents were affected by enhanced UV radiation. The chlorophyll a, b, and total contents were decreased compared with the control values and reduced with the enhanced UV radiation, but the carotenoid was increased compared with the control and also reduced with the enhanced UV radiation. So, the contents of chlorophylls varied considerably. M. parviflora showed the highest constitutive levels of accumulated chlorophyll a, b, and total chlorophyll (0.463, 0.307 and 0.774 mg g(-1) f w) among the investigated plant species. P. major showed the lowest constitutive levels of the chloroplast pigments, 0.0036, 0.0038 and 0.0075 mg g(-1) f w for chlorophyll a, b, and total chlorophyll at UV-365 nm, respectively. The protein content was decreased significantly in both root and shoot systems compared with the control values but, it was increased with increasing wave lengths of UV-radiation of all tested plants. R. vesicarius showed the highest protein contents among the investigated plants; its content was 3.8 mg g(-1) f w at UV-365 nm in shoot system. On the other hand, decreasing ultraviolet wave length induced a highly significant increase in the level of proline in both root and shoot of all

  8. Evidence that COMT genotype and proline interact on negative-symptom outcomes in schizophrenia and bipolar disorder.

    Clelland, C L; Drouet, V; Rilett, K C; Smeed, J A; Nadrich, R H; Rajparia, A; Read, L L; Clelland, J D


    Elevated peripheral proline is associated with psychiatric disorders, and there is evidence that proline is a neuromodulator. The proline dehydrogenase (PRODH) gene, which encodes the enzyme that catalyzes proline catabolism, maps to human chromosome 22q11.2, a region conferring risk of schizophrenia. In the Prodh-null mouse, an interaction between elevated peripheral proline and another 22q11.2 gene, catechol-O-methyltransferase (COMT), on neurotransmission and behavior has been reported. We explored the relationship between fasting plasma proline levels and COMT Val(158)Met genotype on symptoms (positive, negative and total) in schizophrenia patients. In an exploratory study we also examined symptom change in patients with bipolar disorder. There was a significant interaction between peripheral proline and COMT on negative symptoms in schizophrenia (P<0.0001, n=95). In COMT Val/Val patients, high proline was associated with low Scale for the Assessment of Negative Symptom (SANS) scores. In contrast, high proline was associated with high SANS scores in patients carrying a Met allele. The relationship between proline and COMT also appears to modify negative symptoms across psychiatric illness. In bipolar disorder, a significant interaction was also observed on negative-symptom change (P=0.007, n=43). Negative symptoms are intractable and largely unaddressed by current medications. These data indicate a significant interaction between peripheral proline and COMT genotype, influencing negative symptoms in schizophrenia and bipolar disorder. That high proline has converse effects on symptoms by COMT genotype, may have implications for therapeutic decisions.

  9. [b][/b]Impact of copper (Cu at the dose of 50 mg on haematological and biochemical blood parameters in turkeys, and level of Cu accumulation in the selected tissues as a source of information on product safety for consumers

    Bogusław Makarski


    Full Text Available Introduction. The current state-of the art points to a positive impact of copper (Cu supplements on the general health status in poultry. Copper induces beneficial changes in the haematological and biochemical blood parameters. It also displays immunostimulating properties and helps maintain a proper microbiological balance in the digestive tract. [b]Objective[/b]. The objective of this study was to investigate the impact of Cu at the dose of 50 mg/kg BW, administered in organic and inorganic form, on the haematological and biochemical blood parameters and level of Cu bioaccumulation in the liver and pectoral muscle. [b]Materials and method[/b]. The study was carried out on 45 BUT-9 turkeys which had been were reared for 16 weeks. They were divided into 3 experimental groups: I – the control group; II – fed with CuSO[sub]4[/sub] at the dose of 50 mg Cu•dm [sup]-3 [/sup]H[sub]2[/sub]O; III – received a Cu chelate with lysine at the same dose. [b]Results[/b]. The administration of Cu at the dose exceeding the nutritional recommendations did not induce beneficial changes in the examined birds. This indicates that it is not necessary to administer Cu doses higher than the recommended levels. The extent of Cu accumulation in the pectoral muscle increased by 40% compared to the control group, whereas in the liver it was higher by 30–35% than in the birds without Cu administration. The level of Cu in tissues does not pose a risk to consumers. [b]Conclusions[/b]. The supplementation of Cu at the dose of 50 mg has a negative impact on the level of the analyzed parameters. The results of the presented study indicate that the administered Cu dose exceeds birds’ demand for this element.

  10. l-Proline and RNA Duplex m-Value Temperature Dependence.

    Schwinefus, Jeffrey J; Baka, Nadia L; Modi, Kalpit; Billmeyer, Kaylyn N; Lu, Shutian; Haase, Lucas R; Menssen, Ryan J


    The temperature dependence of l-proline interactions with the RNA dodecamer duplex surface exposed after unfolding was quantified using thermal and isothermal titration denaturation monitored by uv-absorbance. The m-value quantifying proline interactions with the RNA duplex surface area exposed after unfolding was measured using RNA duplexes with GC content ranging between 17 and 83%. The m-values from thermal denaturation decreased with increasing GC content signifying increasingly favorable proline interactions with the exposed RNA surface area. However, m-values from isothermal titration denaturation at 25.0 °C were independent of GC content and less negative than those from thermal denaturation. The m-value from isothermal titration denaturation for a 50% GC RNA duplex decreased (became more negative) as the temperature increased and was in nearly exact agreement with the m-value from thermal denaturation. Since RNA duplex transition temperatures increased with GC content, the more favorable proline interactions with the high GC content duplex surface area observed from thermal denaturation resulted from the temperature dependence of proline interactions rather than the RNA surface chemical composition. The enthalpy contribution to the m-value was positive and small (indicating a slight increase in duplex unfolding enthalpy with proline) while the entropic contribution to the m-value was positive and increased with temperature. Our results will facilitate proline's use as a probe of solvent accessible surface area changes during biochemical reactions at different reaction temperatures.

  11. Effect of proline as a nutrient on hypopharyngeal glands during development of Apis mellifera (Hymenoptera: Apidae

    Ali Darvishzadeh


    Full Text Available Proline is known to be an energy source for protein synthesis and appears to have a major role in insect flying metabolism. Insects can detect proline in their food and use it as an energy substrate to start flight and other high energy consuming activities. Honey bee has a feeding preference for nectars with higher concentrations of this amino acid. In this research we present evidence that L-proline can be utilized as a phagostimulant for the honeybee worker (Apis mellifera. We reported the L-proline increase hypopharyngeal glands acini diameter and syrup consumption at the experimental cage. Honeybee workers fed on 1000 ppm treatment prolin consumed 773.9±31.8 ul/bee after 18-days. It is obvious that the honeybee workers consumed 1000 ppm the more than other treatment. The feeding decreased when concentration of L-proline increased to 10000 ppm. The hypopharyngeal glands development increased gradually from honeybee workers emergence and started to decrease after 9 days old. The maximum acini diameter (0.1439±0.001 mm was recorded in the 9th day when newly emerged bees were fed on 1000 ppm proline syrup.

  12. Proline auxotrophy in Sinorhizobium meliloti results in a plant-specific symbiotic phenotype.

    diCenzo, George C; Zamani, Maryam; Cowie, Alison; Finan, Turlough M


    In order to effectively manipulate rhizobium-legume symbioses for our benefit, it is crucial to first gain a complete understanding of the underlying genetics and metabolism. Studies with rhizobium auxotrophs have provided insight into the requirement for amino acid biosynthesis during the symbiosis; however, a paucity of available L-proline auxotrophs has limited our understanding of the role of L-proline biosynthesis. Here, we examined the symbiotic phenotypes of a recently described Sinorhizobium meliloti L-proline auxotroph. Proline auxotrophy was observed to result in a host-plant-specific phenotype. The S. meliloti auxotroph displayed reduced symbiotic capability with alfalfa (Medicago sativa) due to a decrease in nodule mass formed and therefore a reduction in nitrogen fixed per plant. However, the proline auxotroph formed nodules on white sweet clover (Melilotus alba) that failed to fix nitrogen. The rate of white sweet clover nodulation by the auxotroph was slightly delayed, but the final number of nodules per plant was not impacted. Examination of white sweet clover nodules by confocal microscopy and transmission electron microscopy revealed the presence of the S. meliloti proline auxotroph cells within the host legume cells, but few differentiated bacteroids were identified compared with the bacteroid-filled plant cells of WT nodules. Overall, these results indicated that L-proline biosynthesis is a general requirement for a fully effective nitrogen-fixing symbiosis, likely due to a transient requirement during bacteroid differentiation.

  13. Functional characterization and expression analysis of rice δ(1)-pyrroline-5-carboxylate dehydrogenase provide new insight into the regulation of proline and arginine catabolism.

    Forlani, Giuseppe; Bertazzini, Michele; Zarattini, Marco; Funck, Dietmar


    While intracellular proline accumulation in response to various stress conditions has been investigated in great detail, the biochemistry and physiological relevance of proline degradation in plants is much less understood. Moreover, the second and last step in proline catabolism, the oxidation of δ(1)-pyrroline-5-carboxylic acid (P5C) to glutamate, is shared with arginine catabolism. Little information is available to date concerning the regulatory mechanisms coordinating these two pathways. Expression of the gene coding for P5C dehydrogenase was analyzed in rice by real-time PCR either following the exogenous supply of amino acids of the glutamate family, or under hyperosmotic stress conditions. The rice enzyme was heterologously expressed in E. coli, and the affinity-purified protein was thoroughly characterized with respect to structural and functional properties. A tetrameric oligomerization state was observed in size exclusion chromatography, which suggests a structure of the plant enzyme different from that shown for the bacterial P5C dehydrogenases structurally characterized to date. Kinetic analysis accounted for a preferential use of NAD(+) as the electron acceptor. Cations were found to modulate enzyme activity, whereas anion effects were negligible. Several metal ions were inhibitory in the micromolar range. Interestingly, arginine also inhibited the enzyme at higher concentrations, with a mechanism of uncompetitive type with respect to P5C. This implies that millimolar levels of arginine would increase the affinity of P5C dehydrogenase toward its specific substrate. Results are discussed in view of the involvement of the enzyme in either proline or arginine catabolism.

  14. Hot air treatment-induced arginine catabolism is associated with elevated polyamines and proline levels and alleviates chilling injury in postharvest tomato fruit.

    Zhang, Xinhua; Shen, Lin; Li, Fujun; Meng, Demei; Sheng, Jiping


    To understand whether arginine catabolism might be involved in hot air (HA)-induced chilling tolerance mechanism in tomato fruit, we investigated the effect of HA treatment on endogenous arginine catabolism in relation to chilling injury. Tomato fruit were harvested at mature green stage and treated with HA at 38°C for 12 h and then stored at 2°C for 21 days. The effects of HA treatment on fruit chilling injury and gene expression levels or enzyme activity, and metabolites related to arginine catabolism were evaluated. HA treatment reduced the chilling injury symptoms of tomato fruit and enhanced the accumulation of endogenous polyamines, especially putrescine and proline. This accumulation is associated with the increased transcript levels of genes encoding arginase (LeARG1 and LeARG2), arginine decarboxylase (LeADC), ornithine decarboxylase (LeODC) and ornithine aminotransferase (LeOAT) at most sampling times. However, HA treatment had little effect on nitric oxide synthase activity and nitric oxide concentration. These results revealed that the reduction in chilling injury by HA treatment may be due to the accumulation of putrescine and proline induced primarily by activating the catabolism of endogenous arginine. © 2013 Society of Chemical Industry.

  15. Proline iminopeptidase PepI overexpressing Lactobacillus casei as an adjunct starter in Edam cheese

    Navidghasemizad, Sahar; Takala, Timo M; Alatossava, Tapani; Saris, Per EJ


    In this study the growth of genetically modified Lactobacillus casei LAB6, overexpressing proline iminopeptidase PepI and its capacity to increase free proline was investigated during ripening of Edam cheese. The strain successfully survived 12 weeks of ripening period in cheese. The food-grade plasmid pLEB604, carrying the pepI gene, was stable, and PepI enzyme was active in LAB6 cells isolated at different stages of the ripening process. However, HPLC analyses indicated that Lb. casei LAB6 could not increase the amount of free proline in ripened cheese. PMID:23851577

  16. The influence of lactation on L-proline absorption from small intestine in the albino rat.

    Datta, U; Sharma, R K


    Intestinal absorption of L-proline was studied in control and lactating rats from jejunum and ileum by in vivo method and presented per unit dry weight and per unit length of the respective segment. L-proline absorption was found to be significantly reduced in lactating animals as compared to the virgin controls. The results were discussed in light of serosal to mucosal ratio. By in vitro method also jejunal and ileal uptake of L-proline were found to be significantly reduced in lactating animals as compared to the virgin controls.

  17. L-Proline Catalyzed Solvent-Free Knoevenagel Condensation for the Synthesis of 3-Substituted Coumarins

    KARADE Nandkishor. N.; GAMPAWAR Sumit V.; SHINDE Sandeep. V.; JADHAV Wamanrao. N.


    L-Proline was utilized as an efficient organo-catalyst for the environmentally benign synthesis of 3-substituted coumarins by the Knoevenagel condensation of substituted 2-hydroxybenzaldehydes with reactive methylene compounds under solvent free conditions.

  18. Direct asymmetric aldol reaction using MBHA resin-supported peptide containing L-proline unit

    Liang Zhang; Wen Bo Ding; Yong Ping Yu; Hong Bin Zou


    MBHA resin-supported tripeptide catalyst system containing L-proline unit has been developed for use in the direct asymmetric aldol reaction of acetone and aldehydes,which afford the corresponding products with satisfactory isolated yields and enantiomeric excesses.

  19. Novel chiral ionic liquid (CIL) assisted selectivity enhancement to (L)-proline catalyzed asymmetric aldol reactions

    Zhang, Long; Zhang, Haibo; Luo, Huadong; Zhou, Xiaohai; Cheng, Gongzhen [College of Chemistry, Wuhan University (China)


    A significant improvement of the chemical yields (up to 88%), stereoselectivity (> 99:1) and enantiomeric excesses (up to 98%) of (L)-proline catalyzed direct asymmetric aldol reaction was found when proline based chiral ionic liquids (CILs) were added as additives. Different ratios of DMSO/H{sub 2}O as solvent and chiral ionic liquids (CILs) with chiral cations of different chain length were investigated. (author)

  20. The dominant glutamic acid metabolic flux to produce γ-amino butyric acid over proline in Nicotiana tabacum leaves under water stress relates to its significant role in antioxidant activity.

    Liu, Cuili; Zhao, Li; Yu, Guanghui


    γ-Amino butyric acid (GABA) and proline play a crucial role in protecting plants during various environmental stresses. Their synthesis is from the common precursor glutamic acid, which is catalyzed by glutamate decarboxylase and Δ(1) -pyrroline-5-carboxylate synthetase respectively. However, the dominant pathway under water stress has not yet been established. To explore this, excised tobacco leaves were used to simulate a water-stress condition. The results showed GABA content was much higher than that of proline in leaves under water-deficit and non-water-deficit conditions. Specifically, the amount of GABA significantly increased compared to proline under continuous water loss for 16 h, indicating that GABA biosynthesis is the dominant pathway from glutamic acid metabolism under these conditions. Quantitative reverse transcription polymerase chain reaction and protein Western gel-blot analysis further confirmed this. To explore the function of GABA accumulation, a system producing superoxide anion (O(2) (-) ), peroxide hydrogen (H(2) O(2) ), and singlet oxygen ((1) O(2) ) was employed to investigate the scavenging role on free-radical production. The results demonstrated that the scavenging ability of GABA for O(2) (-) , H(2) O(2) , and (1) O(2) was significantly higher than that of proline. This indicated that GABA acts as an effective osmolyte to reduce the production of reactive oxygen species under water stress.

  1. Variação do teor de prolina em folhas de feijão em função da disponibilidade de água no solo Changes in proline content in leaves of Phaseolus vulgaris L. in response to water stress

    Haiko Enok Sawazaki


    Full Text Available Para verificar o comportamento de cultivares e linhagens de feijão quanto à capacidade de acumular prolina livre em suas folhas em condições de escassez de água no solo, foram utilizados dezoito cultivares e duas linhagens de feijão, desenvolvidos em casa de vegetação. Desse material, foram amostradas folhas primárias, de acordo com os tratamentos: a irrigado diariamente; b onze e quinze dias sem irrigação; e c plantas reidratadas após onze dias sem irrigação, com o objetivo de avaliar o teor de prolina. Os resultados obtidos mostraram diferenças no teor de prolina e na capacidade de acúmulo desse aminoácido em função de cultivares e tratamentos. 'Jalo', 'Roseli' e 'Roxão Lustroso' acumularam maiores quantidades de prolina (> 7 micromoles/grama de matéria seca, enquanto 'Moruna' e 'Curitibano-Bairro das Palmeiras' mostraram os menores acúmulos (The objective of this paper was to verify the accumulation of free proline in leaves of beans (Phaseolus vulgaris L. when subjected to water stress. Leaves samples were taken at 11 and 15 days after starting the water stress and 4 days after irrigation to estimate the proline accumulation. The results obtained showed differences among bean cultivars in the proline content and the capacity for accumulation of this aminoacid under 15 days of water stress. 'Jalo', 'Roseli', and 'Roxão Lustroso' were cultivars with the highest proline accumulation (> 7 mmol/g DW whereas 'Moruna' and 'Curitibano Bairro das Palmeiras' were cultivars with the lowest proline content (< 3 mmol/g DW, after 15 days of water stress. The addition of water after 11 days of water stress showed proline content similar to the irrigated plants. The materials studied were classified as a function of their capacity for proline accumulation following 15 days of water stress. This classification represents differences in drought resistance if a higher proline contents is considered a measurement of the plant adaptation

  2. Proline dehydrogenase from Thermus thermophilus does not discriminate between FAD and FMN as cofactor

    Huijbers, Mieke M. E.; Martínez-Júlvez, Marta; Westphal, Adrie H.; Delgado-Arciniega, Estela; Medina, Milagros; van Berkel, Willem J. H.


    Flavoenzymes are versatile biocatalysts containing either FAD or FMN as cofactor. FAD often binds to a Rossmann fold, while FMN prefers a TIM-barrel or flavodoxin-like fold. Proline dehydrogenase is denoted as an exception: it possesses a TIM barrel-like fold while binding FAD. Using a riboflavin auxotrophic Escherichia coli strain and maltose-binding protein as solubility tag, we produced the apoprotein of Thermus thermophilus ProDH (MBP-TtProDH). Remarkably, reconstitution with FAD or FMN revealed that MBP-TtProDH has no preference for either of the two prosthetic groups. Kinetic parameters of both holo forms are similar, as are the dissociation constants for FAD and FMN release. Furthermore, we show that the holo form of MBP-TtProDH, as produced in E. coli TOP10 cells, contains about three times more FMN than FAD. In line with this flavin content, the crystal structure of TtProDH variant ΔABC, which lacks helices αA, αB and αC, shows no electron density for an AMP moiety of the cofactor. To the best of our knowledge, this is the first example of a flavoenzyme that does not discriminate between FAD and FMN as cofactor. Therefore, classification of TtProDH as an FAD-binding enzyme should be reconsidered. PMID:28256579

  3. Growth and characterization of new semi-organic L-proline strontium chloride monohydrate single crystals

    Gupta, Manoj K.; Sinha, Nidhi; Kumar, Binay


    The present communication deals with the synthesis, single crystal growth and characterization of a new nonlinear optical material L-proline strontium chloride monohydrate ( L-PSCM). Single crystals have been grown using the slow solvent evaporation technique. Single crystal XRD analysis confirmed that the crystal belongs to the orthorhombic structure with lattice parameter a=6.6966(3) Å, b=12.4530(5) Å, c=15.2432(5) Å and space group P2 12 12 1. Presence of various functional groups in L-PSCM and protonation of the ions were confirmed by Fourier transform infrared spectroscopy (FT-IR) analysis. The melting point of the single crystal was found to be 126 °C using DSC. Ultraviolet-visible spectral analyses showed that the crystal has low UV cut-off at 226 nm combined with very good transparency of 90% in a wide range. The optical band gap was estimated to be 5.82 eV. Capacitance and dielectric-loss measurements were carried out at different temperatures in the frequency range 1 kHz-2 MHz. The dielectric constant and loss factor were found to be 21 and 0.03 at 1 kHz at room temperature, respectively. Microhardness mechanical studies show that hardness number ( Hv) increases with load for L-PSCM single crystals the by Vickers microhardness method. Second harmonic generation (SHG) efficiency was found to be 0.078 times the value of KDP.

  4. Proline content and protein patterns in Eucalyptus grandis shoots submitted to high and low temperature shocks

    Gustavo M. Souza


    Full Text Available Proline content and protein patterns changes in response to temperature shocks of both acclimated and non acclimated E. grandis shoots cultivated in vitro were investigated. Analysis of soluble proteins through SDS-PAGE and proline were carried out after 12h at 12ºC (cold acclimation or 33ºC (heat acclimation, and immediately after temperature shocks at 41ºC and 0ºC. Analyses were also performed 24h after the temperature shocks (recovery period. Temperature treatment at 0ºC did not change soluble protein patterns both in acclimatized and non acclimatized plants, whereas cold temperature induced high proline levels, which kept relatively high after recovery period. Three novel, probably HSPs, proteins (90.5, 75 and 39 kDa were observed in both acclimated and non acclimated plants submitted to high temperatures. Plants exposed at 41ºC were able to recover from heat shocks after 24h, whereas they did not recover completely from cold shocks. The effect of the acclimation period on the recovering (homeostasis varied depending on the parameter evaluated, and type and duration of the temperature shock.As respostas às mudanças de temperatura de plantas aclimatadas e não aclimatadas de E. grandis cultivadas in vitro foram avaliadas considerando alterações dos níveis de prolina e proteínas solúveis totais. Análises de proteínas solúveis através de SDS-PAGE e prolina foram realizadas após 12h a 12ºC (aclimatação ao frio ou a 33ºC (aclimatação ao calor, e imediatamente depois dos choques térmicos a 41ºC e 0ºC. Análises também foram realizadas após um período de 24h depois dos choques térmicos (período de recuperação. O tratamento de temperatura a 0ºC não alterou o padrão de proteínas nas plantas aclimatadas e não aclimatadas, entretanto a temperatura baixa induziu altos níveis de prolina, que se mantiveram relativamente altos após o período de recuperação. Três novas proteínas (90,5, 75 e 39 kDa, provavelmente

  5. The mammalian profilin isoforms display complementary affinities for PIP2 and proline-rich sequences.

    Lambrechts, A; Verschelde, J L; Jonckheere, V; Goethals, M; Vandekerckhove, J; Ampe, C


    We present a study on the binding properties of the bovine profilin isoforms to both phosphatidylinositol 4,5-bisphosphate (PIP2) and proline-rich peptides derived from vasodilator-stimulated phosphoprotein (VASP) and cyclase-associated protein (CAP). Using microfiltration, we show that compared with profilin II, profilin I has a higher affinity for PIP2. On the other hand, fluorescence spectroscopy reveals that proline-rich peptides bind better to profilin II. At micromolar concentrations, profilin II dimerizes upon binding to proline-rich peptides. Circular dichroism measurements of profilin II reveal a significant conformational change in this protein upon binding of the peptide. We show further that PIP2 effectively competes for binding of profilin I to poly-L-proline, since this isoform, but not profilin II, can be eluted from a poly-L-proline column with PIP2. Using affinity chromatography on either profilin isoform, we identified profilin II as the preferred ligand for VASP in bovine brain extracts. The complementary affinities of the profilin isoforms for PIP2 and the proline-rich peptides offer the cell an opportunity to direct actin assembly at different subcellular localizations through the same or different signal transduction pathways.

  6. Stereochemical Sequence Ion Selectivity: Proline versus Pipecolic-acid-containing Protonated Peptides

    Abutokaikah, Maha T.; Guan, Shanshan; Bythell, Benjamin J.


    Substitution of proline by pipecolic acid, the six-membered ring congener of proline, results in vastly different tandem mass spectra. The well-known proline effect is eliminated and amide bond cleavage C-terminal to pipecolic acid dominates instead. Why do these two ostensibly similar residues produce dramatically differing spectra? Recent evidence indicates that the proton affinities of these residues are similar, so are unlikely to explain the result [Raulfs et al., J. Am. Soc. Mass Spectrom. 25, 1705-1715 (2014)]. An additional hypothesis based on increased flexibility was also advocated. Here, we provide a computational investigation of the "pipecolic acid effect," to test this and other hypotheses to determine if theory can shed additional light on this fascinating result. Our calculations provide evidence for both the increased flexibility of pipecolic-acid-containing peptides, and structural changes in the transition structures necessary to produce the sequence ions. The most striking computational finding is inversion of the stereochemistry of the transition structures leading to "proline effect"-type amide bond fragmentation between the proline/pipecolic acid-congeners: R (proline) to S (pipecolic acid). Additionally, our calculations predict substantial stabilization of the amide bond cleavage barriers for the pipecolic acid congeners by reduction in deleterious steric interactions and provide evidence for the importance of experimental energy regime in rationalizing the spectra.

  7. Stereochemical Sequence Ion Selectivity: Proline versus Pipecolic-acid-containing Protonated Peptides

    Abutokaikah, Maha T.; Guan, Shanshan; Bythell, Benjamin J.


    Substitution of proline by pipecolic acid, the six-membered ring congener of proline, results in vastly different tandem mass spectra. The well-known proline effect is eliminated and amide bond cleavage C-terminal to pipecolic acid dominates instead. Why do these two ostensibly similar residues produce dramatically differing spectra? Recent evidence indicates that the proton affinities of these residues are similar, so are unlikely to explain the result [Raulfs et al., J. Am. Soc. Mass Spectrom. 25, 1705-1715 (2014)]. An additional hypothesis based on increased flexibility was also advocated. Here, we provide a computational investigation of the "pipecolic acid effect," to test this and other hypotheses to determine if theory can shed additional light on this fascinating result. Our calculations provide evidence for both the increased flexibility of pipecolic-acid-containing peptides, and structural changes in the transition structures necessary to produce the sequence ions. The most striking computational finding is inversion of the stereochemistry of the transition structures leading to "proline effect"-type amide bond fragmentation between the proline/pipecolic acid-congeners: R (proline) to S (pipecolic acid). Additionally, our calculations predict substantial stabilization of the amide bond cleavage barriers for the pipecolic acid congeners by reduction in deleterious steric interactions and provide evidence for the importance of experimental energy regime in rationalizing the spectra.

  8. Synthesis and biological activity of peptide proline-boronic acids as proteasome inhibitors.

    Han, Liqiang; Wen, Yanzhao; Li, Ridong; Xu, Bo; Ge, Zemei; Wang, Xin; Cheng, Tieming; Cui, Jingrong; Li, Runtao


    On the basis of the application of proline-boronic acid as pharmacophore in the kinase inhibitors and our previous research results, using proline-boronic acid as warhead, two series of peptide proline-boronic acids, dipeptide proline-boronic acids (I) and tripeptide proline-boronic acids (II), were designed and synthesized. All the synthesized compounds were first evaluated for their biological activity against MGC803 cell, and then, the best compound II-7 was selected to test its anti-tumor spectrum on six human tumor cell lines and proteasome inhibition against three subunits. The results indicated that series II have much better biological activities than series I. The compound II-7 exhibited not only excellent biological activities with IC50 values of nM level in both cell and proteasome models, but also much better subunit selectivity. Thus, proline-boronic acid as warhead is reasonable in the design of proteasome inhibitors. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Charge accumulation in lossy dielectrics: a review

    Rasmussen, Jørgen Knøster; McAllister, Iain Wilson; Crichton, George C


    such that the material parameters which influence charge accumulation are clearly identified; viz. the conductivity, permittivity and dimensions of the insulating media. The two former parameters, together with the applied voltage, govern both the magnitude and polarity of the accumulated charge....

  10. The proline-rich motif of the proDer p 3 allergen propeptide is crucial for protease-protease interaction.

    Marie-Eve Dumez

    Full Text Available The majority of proteases are synthesized in an inactive form, termed zymogen, which consists of a propeptide and a protease domain. The propeptide is commonly involved in the correct folding and specific inhibition of the enzyme. The propeptide of the house dust mite allergen Der p 3, NPILPASPNAT, contains a proline-rich motif (PRM, which is unusual for a trypsin-like protease. By truncating the propeptide or replacing one or all of the prolines in the non-glycosylated zymogen with alanine(s, we demonstrated that the full-length propeptide is not required for correct folding and thermal stability and that the PRM is important for the resistance of proDer p 3 to undesired proteolysis when the protein is expressed in Pichia pastoris. Additionally, we followed the maturation time course of proDer p 3 by coupling a quenched-flow assay to mass spectrometry analysis. This approach allowed to monitor the evolution of the different species and to determine the steady-state kinetic parameters for activation of the zymogen by the major allergen Der p 1. This experiment demonstrated that prolines 5 and 8 are crucial for proDer p 3-Der p 1 interaction and for activation of the zymogen.

  11. Disruption of AtWNK8 Enhances Tolerance of Arabidopsis to Salt and Osmotic Stresses via Modulating Proline Content and Activities of Catalase and Peroxidase

    Hong Liao


    Full Text Available With no lysine kinases (WNKs play important roles in plant growth and development. However, its role in salt and osmotic stress tolerance is unclear. Here, we report that AtWNK8 is mainly expressed in primary root, hypocotyl, stamen and pistil and is induced by NaCl and sorbitol treatment. Compared to the wild-type, the T-DNA knock-out wnk8 mutant was more tolerant to severe salinity and osmotic stresses, as indicated by 27% and 198% more fresh weight in the NaCl and sorbitol treatment, respectively. The wnk8 mutant also accumulated 1.43-fold more proline than the wild-type in the sorbitol treatment. Under NaCl and sorbitol stresses, catalase (CAT activity in wnk8 mutant was 1.92- and 3.7-times of that in Col-0, respectively. Similarly, under salt and osmotic stress conditions, peroxidase (POD activities in wnk8 mutant were 1.81- and 1.58-times of that in Col-0, respectively. Taken together, we revealed that maintaining higher CAT and POD activities might be one of the reasons that the disruption of AtWNK8 enhances the tolerance to salt stress, and accumulating more proline and higher activities of CAT and POD might result in the higher tolerance of WNK8 to osmotic stress.

  12. Studying of Salinity Stress Effect on Germination, Proline, Sugar, Protein, Lipid and Chlorophyll Content in Purslane (Portulaca oleracea L. Leaves

    Parvaneh Rahdari


    Full Text Available Approximately 30% of country regions are under influence of salt and one of the main important methods for increasing of hectare performance in salt soils, using of resistant plant against with salt such as medicinal annual plant of Purslane (Portulaca oleraceae L..Studying of the salt stress effect on germination and some physiological parameters in Purslane was carried out in completely random framework and with three replications and under controlled conditions. In this study, salinity was imposed by sodium chloride (NaCl and in five levels of framework included of Control (distilled water, 50, 100, 150 and 200 Mmol from NaCl. Studied physiological parameters included of proline, sugar, protein, lipid and chlorophyll in Purslane leaves.In relation with observed germination that increasing of salt level, germination rate was reduced that the least germination rate in 200 Mmol and the highest germination was observed in control which in statistical level 0.01 showed meaningful difference. Also, results showed an increasing of leaf proline and sugar that was synchronize with increasing of sodium chloride, this increasing in statistical level of P<0.01 was meaningful, while blank had the least rate and salt level 200 Mmol was observed. Synchronize with increasing of salinity level, lipid and protein concentration decreasing that was meaningful (P<0.01 which highest of leaf lipid and protein content in control level and least concentration was observed in 200 Mmol of salinity level. Also, salinity stress due to increasing of chlorophyll a and b levels in Purslane leaves which this increasing in leaf chlorophyll b concentrations in 0.01 statistical level was meaningful and the least level chlorophyll a and b in blank and the highest level of chlorophyll a and b by order in 150 and 200 Mmol of NaCl was observed. The results showed that plant for resistance against salinity increasing of proline, sugar and chlorophyll content in leaves are useful for

  13. L-Proline catalyzed aldol reactions between acetone and aldehydes in supercritical fluids:An environmentally friendly reaction procedure


    The direct asymmetric aldol reaction between various aldehydes and acetone catalyzed by L-proline catalyst was successfully carried out in supercritical CO2 (scCO2) and 1,1,1,2-tetrafluoroethane (R-134a) fluids.The enantioselectivity of 84% ee to the targeted product was achieved under 20 MPa,40 °C,and 15 mol% of the catalyst in supercritical CO2 (scCO2) fluid.The effects of reaction parameters,such as temperature,pressure,catalyst loading and different substituted aldehydes on both enantioselectivity and aldol yield were discussed.The titled reaction was also performed in 1,1,1,2-tetrafluoroethane,and the obtained results were compared with those in scCO2.This new reaction procedure provides an environmental asymmetric aldol reaction system as compared with that in organic solvents.

  14. Proline Metabolism is Essential for Trypanosoma brucei brucei Survival in the Tsetse Vector

    Mantilla, Brian S.; Dyer, Naomi A.; Biran, Marc; Bringaud, Frédéric; Lehane, Michael J.; Acosta-Serrano, Alvaro


    Adaptation to different nutritional environments is essential for life cycle completion by all Trypanosoma brucei sub-species. In the tsetse fly vector, L-proline is among the most abundant amino acids and is mainly used by the fly for lactation and to fuel flight muscle. The procyclic (insect) stage of T. b. brucei uses L-proline as its main carbon source, relying on an efficient catabolic pathway to convert it to glutamate, and then to succinate, acetate and alanine as the main secreted end products. Here we investigated the essentiality of an undisrupted proline catabolic pathway in T. b. brucei by studying mitochondrial Δ1-pyrroline-5-carboxylate dehydrogenase (TbP5CDH), which catalyzes the irreversible conversion of gamma-glutamate semialdehyde (γGS) into L-glutamate and NADH. In addition, we provided evidence for the absence of a functional proline biosynthetic pathway. TbP5CDH expression is developmentally regulated in the insect stages of the parasite, but absent in bloodstream forms grown in vitro. RNAi down-regulation of TbP5CDH severely affected the growth of procyclic trypanosomes in vitro in the absence of glucose, and altered the metabolic flux when proline was the sole carbon source. Furthermore, TbP5CDH knocked-down cells exhibited alterations in the mitochondrial inner membrane potential (ΔΨm), respiratory control ratio and ATP production. Also, changes in the proline-glutamate oxidative capacity slightly affected the surface expression of the major surface glycoprotein EP-procyclin. In the tsetse, TbP5CDH knocked-down cells were impaired and thus unable to colonize the fly’s midgut, probably due to the lack of glucose between bloodmeals. Altogether, our data show that the regulated expression of the proline metabolism pathway in T. b. brucei allows this parasite to adapt to the nutritional environment of the tsetse midgut. PMID:28114403

  15. Proline and Glutamine Improvein vitro Callus Induction and Subsequent Shooting in Rice

    Bhausaheb PAWAR; Prashant KALE; Jyoti BAHURUPE; Ashok JADHAV; Anil KALE; Sharad PAWAR


    This study was conducted to evaluate the effects of proline and glutamine onin vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four ricegenotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency (83.2%) was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency (83.2%), whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used forAgrobacterium mediated genetic transformation in rice.

  16. Sinorhizobium meliloti chemoreceptor McpU mediates chemotaxis toward host plant exudates through direct proline sensing.

    Webb, Benjamin A; Hildreth, Sherry; Helm, Richard F; Scharf, Birgit E


    Bacterial chemotaxis is an important attribute that aids in establishing symbiosis between rhizobia and their legume hosts. Plant roots and seeds exude a spectrum of molecules into the soil to attract their bacterial symbionts. The alfalfa symbiont Sinorhizobium meliloti possesses eight chemoreceptors to sense its environment and mediate chemotaxis toward its host. The methyl accepting chemotaxis protein McpU is one of the more abundant S. meliloti chemoreceptors and an important sensor for the potent attractant proline. We established a dominant role of McpU in sensing molecules exuded by alfalfa seeds. Mass spectrometry analysis determined that a single germinating seed exudes 3.72 nmol of proline, producing a millimolar concentration near the seed surface which can be detected by the chemosensory system of S. meliloti. Complementation analysis of the mcpU deletion strain verified McpU as the key proline sensor. A structure-based homology search identified tandem Cache (calcium channels and chemotaxis receptors) domains in the periplasmic region of McpU. Conserved residues Asp-155 and Asp-182 of the N-terminal Cache domain were determined to be important for proline sensing by evaluating mutant strains in capillary and swim plate assays. Differential scanning fluorimetry revealed interaction of the isolated periplasmic region of McpU (McpU40-284) with proline and the importance of Asp-182 in this interaction. Using isothermal titration calorimetry, we determined that proline binds with a Kd (dissociation constant) of 104 μM to McpU40-284, while binding was abolished when Asp-182 was substituted by Glu. Our results show that McpU is mediating chemotaxis toward host plants by direct proline sensing.

  17. [Agrobacterium-mediated sunflower transformation (Helianthus annuus L.) in vitro and in Planta using strain of LBA4404 harboring binary vector pBi2E with dsRNA-suppressor proline dehydrogenase gene].

    Tishchenko, E N; Komisarenko, A G; Mikhal'skaia, S I; Sergeeva, L E; Adamenko, N I; Morgun, B V; Kochetov, A V


    To estimate the efficiency of proline dehydrogenase gene suppression towards increasing of sunflower (Helianthus annuus L.) tolerance level to water deficit and salinity, we employed strain LBA4404 harboring pBi2E with double-stranded RNA-suppressor, which were prepared on basis arabidopsis ProDH1 gene. The techniques of Agrobacterium-mediated transformation in vitro and in planta during fertilization sunflower have been proposed. There was shown the genotype-depended integration of T-DNA in sunflower genome. PCR-analysis showed that ProDH1 presents in genome of inbred lines transformed in planta, as well as in T1- and T2-generations. In trans-genic regenerants the essential accumulation of free L-proline during early stages of in vitro cultivation under normal conditions was shown. There was established the essential accumulation of free proline in transgenic regenerants during cultivation under lethal stress pressure (0.4 M mannitol and 2.0% sea water salts) and its decline upon the recovery period. These data are declared about effectiveness of suppression of sunflower ProDH and gene participation in processes connected with osmotolerance.

  18. Assessment of heavy metal accumulation and performance Of some physiological parameters in zea mays l. And Vic1a faba l. Grown on soil amended by sewage Sludge resulting from sewage water Treatment in the state of qatar

    Mazen, A. M. A.


    Pot experiment was conducted to explore extent of concerns regarding agricultural use of sewage sludge for crop production in the state of Qatar. Extent of heavy metal accumulation and its subsequent impact on physiological performance in Zea mays and Viciafaba plants grown on sludge - amended soils was explored. Analysis revealed that all tested heavy metals were several times higher in pure sludge compared to pure garden soil. Plants grown in sludge -mixed soil accumulated large amounts of ...

  19. Asymmetric Aldol Reaction Catalyzed by L-Proline and Achiral Thiourea Fluoroboric Acid Salt

    Cho, Eun; Lee, Haney; Kim, Taek Hyeon [Chonnam National University, Gwangju (Korea, Republic of)


    Considering its ready availability and low cost, L-proline would be the first choice catalyst for preparing aldol adducts with high diastereo- and enantioselectivity. However, proline presents some major drawbacks, including poor performance in direct aldol reactions with aromatic aldehydes, limited solubility, and reactivity in nonpolar organic solvents, and side reactions that make using high catalyst loadings necessary to reach satisfactory conversions. Therefore, numerous proline-modified organo catalysts such as prolinamides, proline thioamides, sulfonamides, chiral amines, and organic salts have been designed for direct aldol reactions. An alternative is to add a readily available additive to the reactions containing proline. This last approach is clearly advantageous in avoiding tedious chemical syntheses of organo catalysts and would ultimately allow the construction of libraries of catalyst protocols by simply changing the additive. Acid additives can influence the outcome of enamine mediated reactions; however, only a few screening studies of acid additions to thiourea organo catalysts are available in the literature. The reaction between cyclohexanone and 4-nitrobenzaldehyde was selected as a standard model reaction for screening of more effective acid additives to thiourea.

  20. Use of prolines for improving growth and other properties of plants and algae

    Unkefer, Pat J.; Knight, Thomas J.; Martinez, Rodolfo A.


    Increasing the concentration of prolines, such as 2-hydroxy-5-oxoproline, in the foliar portions of plants has been shown to cause an increase in carbon dioxide fixation, growth rate, dry weight, nutritional value (amino acids), nodulation and nitrogen fixation, photosynthetically derived chemical energy, and resistance to insect pests over the same properties for wild type plants. This can be accomplished in four ways: (1) the application of a solution of the proline directly to the foliar portions of the plant by spraying these portions; (2) applying a solution of the proline to the plant roots; (3) genetically engineering the plant and screening to produce lines that over-express glutamine synthetase in the leaves which gives rise to increased concentration of the metabolite, 2-hydroxy-5-oxoproline (this proline is also known as 2-oxoglutaramate); and (4) impairing the glutamine synthetase activity in the plant roots which causes increased glutamine synthetase activity in the leaves which gives rise to increased concentration of 2-hydroxy-5-oxoproline. Prolines have also been found to induce similar effects in algae.

  1. Use of prolines for improving growth and other properties of plants and algae

    Unkefer, Pat J. (Los Alamos, NM); Knight, Thomas J. (Portland, ME); Martinez, Rodolfo A. (Santa Fe, NM)


    Increasing the concentration of prolines, such as 2-hydroxy-5-oxoproline, in the foliar portions of plants has been shown to cause an increase in carbon dioxide fixation, growth rate, dry weight, nutritional value (amino acids), nodulation and nitrogen fixation, photosynthetically derived chemical energy, and resistance to insect pests over the same properties for wild type plants. This can be accomplished in four ways: (1) the application of a solution of the proline directly to the foliar portions of the plant by spraying these portions; (2) applying a solution of the proline to the plant roots; (3) genetically engineering the plant and screening to produce lines that over-express glutamine synthetase in the leaves which gives rise to increased concentration of the metabolite, 2-hydroxy-5-oxoproline (this proline is also known as 2-oxoglutaramate); and (4) impairing the glutamine synthetase activity in the plant roots which causes increased glutamine synthetase activity in the leaves which gives rise to increased concentration of 2-hydroxy-5-oxoproline. Prolines have also been found to induce similar effects in algae.

  2. Use of prolines for improving growth and other properties of plants and algae

    Unkefer, Pat J. (Los Alamos, NM); Knight, Thomas J. (Portland, ME); Martinez, Rodolfo A. (Santa Fe, NM)


    Increasing the concentration of prolines such as 2-hydroxy-5-oxoproline, in the foliar portions of plants has been shown to cause an increase in carbon dioxide fixation, growth rate, dry weight, nutritional value (amino acids), nodulation and nitrogen fixation, photosynthetically derived chemical energy, and resistance to insect pests over the same properties for wild type plants. This can be accomplished in four ways: (1) the application of a solution of the proline directly to the foliar portions of the plant by spraying these portions; (2) applying a solution of the proline to the plant roots; (3) genetically engineering the plant and screening to produce lines that overexpress glutamine synthetase in the leaves which gives rise to increased concentration of the metabolite, 2-hydroxy-5-oxoproline (this proline is also known as 2-oxoglutaramnate); and (4) impairing the glutamine synthetase activity in the plant roots which causes increased glutamine synthetase activity in the leaves which gives rise to increased concentration of 2-hydroxy-5-oxoproline. Prolines have also been found to induce similar effects in algae.

  3. Hydroxyproline and proline inhibit α-amylase from isolated barley aleurone layers

    Craig C. Freudenrich


    Full Text Available Previously, we reported that 1 mM hydroxyproline appeared to inhibit the gibberellic acid-induced release of α-amylase from isolated Hordeum vulgare L. cv. Himalaya aleurone layers into an incubation medium. Here, we report our attempts to determine the mechanism(s for this inhibition and whether this inhibition can be caused by other proline analogues. Both 1 mM hydroxyproline and proline inhibited extracellular a-amylase activity without affecting its intracellular activity. This suggested that neither hydroxyproline nor proline impaired the release of a-amylase. Lineweaver-Burk plots revealed that both hydroxyproline and proline uncompetitively inhibited α-amy-lase. Thus, the inhibition is probably an assay artifact resulting from the formation of an enzyme-substrate-hydroxyproline or -proline complex. Because azetidine-2-carboxylic acid, glutamic acid and pipecolic acid did not inhibit extracellular α-amylase activity, the uncompetitive inhibition of a-amylase must be unique to imino aicids as well as their precursors and derivatives which possess a five membered ring.

  4. Water-Deficit Tolerance in Sweet Potato [Ipomoea batatas (L. Lam.] by Foliar Application of Paclobutrazol: Role of Soluble Sugar and Free Proline

    Suravoot Yooyongwech


    Full Text Available The objective of this study was to elevate water deficit tolerance by improving soluble sugar and free proline accumulation, photosynthetic pigment stabilization, photosynthetic abilities, growth performance and storage root yield in sweet potato cv. ‘Tainung 57’ using a foliar application of paclobutrazol (PBZ. The experiment followed a Completely Randomized Block Design with four concentrations of PBZ: 0 (control, 17, 34, and 51 μM before exposure to 47.5% (well irrigation, 32.3% (mild water deficit or 17.5% (severe water deficit soil water content. A sweet potato cultivar, ‘Japanese Yellow’, with water deficit tolerance attributes was the positive check in this study. Total soluble sugar content (sucrose, glucose, and fructose increased by 3.96-folds in ‘Tainung 57’ plants treated with 34 μM PBZ grown under 32.3% soil water content (SWC compared to the untreated plants, adjusting osmotic potential in the leaves and controlling stomatal closure (represented by stomatal conductance and transpiration rate. In addition, under the same treatment, free proline content (2.15 μmol g-1 FW increased by 3.84-folds when exposed to 17.5% SWC. PBZ had an improved effect on leaf size, vine length, photosynthetic pigment stability, chlorophyll fluorescence, and net photosynthetic rate; hence, delaying wilting symptoms and maintaining storage root yield (26.93 g plant-1 at the harvesting stage. A positive relationship between photon yield of PSII (ΦPSII and net photosynthetic rate was demonstrated (r2 = 0.73. The study concludes that soluble sugar and free proline enrichment in PBZ-pretreated plants may play a critical role as major osmoprotectant to control leaf osmotic potential and stomatal closure when plants were subjected to low soil water content, therefore, maintaining the physiological and morphological characters as well as storage root yield.

  5. The Antiproton Accumulator (AA)


    Section 06 - 08*) of the AA where the dispersion (and hence the horizontal beam size) is large. One can distinguish (left to right): A vacuum-tank, two bending magnets (BST06 and BST07 in blue) with a quadrupole (QDN07, in red) in between, another vacuum-tank, a wide quadrupole (QFW08) and a further tank . The tanks are covered with heating tape for bake-out. The tank left of BST06 contained the stack core pickup for stochastic cooling (see 7906193, 7906190, 8005051), the two other tanks served mainly as vacuum chambers in the region where the beam was large. Peter Zettwoch works on BST06. *) see: H. Koziol, Antiproton Accumulator Parameter List, PS/AA/Note 84-2 (1984)

  6. Synthesis, Surface Active Properties and Cytotoxicity of Sodium N-Acyl Prolines.

    Sreenu, Madhumanchi; Narayana Prasad, Rachapudi Badari; Sujitha, Pombala; Kumar, Chityal Ganesh


    Sodium N-acyl prolines (NaNAPro) were synthesized using mixture of fatty acids obtained from coconut, palm, karanja, Sterculia foetida and high oleic sunflower oils via Schotten-Baumann reaction in 58-75% yields to study the synergetic effect of mixture of hydrophobic fatty acyl functionalities like saturation, unsaturation and cyclopropene fatty acids with different chain lengths and aliphatic hetero cyclic proline head group on their surface and cytotoxicity activities. The products were characterized by chromatographic and spectral techniques. The synthesized products were evaluated for their surface active properties such as surface tension, wetting power, foaming characteristics, emulsion stability, calcium tolerance, critical micelle concentration (CMC) and thermodynamic properties. The results revealed that all the products exhibited superior surface active properties like CMC, calcium tolerance and emulsion stability as compared to the standard surfactant, sodium lauryl sulphate (SLS). In addition, palm, Sterculia foetida and high oleic sunflower fatty N-acyl prolines exhibited promising cytotoxicity against different tumor cell lines.

  7. Growth and characterization of new semi-organic L-proline strontium chloride monohydrate single crystals

    Gupta, Manoj K. [Crystal Lab, Department of Physics and Astrophysics, University of Delhi, Delhi 7 (India); Sinha, Nidhi [Department of Electronics, SGTB Khalsa College, University of Delhi, Delhi 7 (India); Kumar, Binay, E-mail: [Crystal Lab, Department of Physics and Astrophysics, University of Delhi, Delhi 7 (India)


    The present communication deals with the synthesis, single crystal growth and characterization of a new nonlinear optical material L-proline strontium chloride monohydrate (L-PSCM). Single crystals have been grown using the slow solvent evaporation technique. Single crystal XRD analysis confirmed that the crystal belongs to the orthorhombic structure with lattice parameter a=6.6966(3) A, b=12.4530(5) A, c=15.2432(5) A and space group P2{sub 1}2{sub 1}2{sub 1}. Presence of various functional groups in L-PSCM and protonation of the ions were confirmed by Fourier transform infrared spectroscopy (FT-IR) analysis. The melting point of the single crystal was found to be 126 {sup o}C using DSC. Ultraviolet-visible spectral analyses showed that the crystal has low UV cut-off at 226 nm combined with very good transparency of 90% in a wide range. The optical band gap was estimated to be 5.82 eV. Capacitance and dielectric-loss measurements were carried out at different temperatures in the frequency range 1 kHz-2 MHz. The dielectric constant and loss factor were found to be 21 and 0.03 at 1 kHz at room temperature, respectively. Microhardness mechanical studies show that hardness number (H{sub v}) increases with load for L-PSCM single crystals the by Vickers microhardness method. Second harmonic generation (SHG) efficiency was found to be 0.078 times the value of KDP.

  8. Heavy Metal Accumulation and Ecological Responses of Turfgrass to Rubbish Compost with EDTA Addition

    Li-An DUO; Yu-Bao GAO; Shu-Lan ZHAO


    concentrations >20 mmol/kg inhibited these parameters significantly for both species. Moreover, EDTA increased the chlorophyll and proline content at all concentrations tested. On the basis of the synthetic remediation index, the optimal EDTA concentration for turfgrass remediation of heavy metals in compost is approximately 10 mmol/kg.

  9. 颗粒离散元模拟堆积碎石土变形的参数灵敏度分析%Particles discrete element simulation accumulation of gravel soil deformation parameter sensitivity analysis

    董辉; 马一跃; 傅鹤林; 王智超; 陈铖


    The author uses the granular discrete element method to simulate the arbitrary shape stone and calibrate the mesoscopic parameters of gravel soil which was mainly constituted by weathering , unloading ,alluvial ,deluvial ,etc .Calibration is based on indoor triaxial compression experiments meas‐ured data at the same time considering the scale effect of sample .T his paper analyzed the sensitivity of the mesoscopic parameters w hich affection the accumulation of gravel soil macro deformation characteris‐tics through the virtual experiment .Studies have shown that :① The mesoscopic parameters of gravel soils based on indoor experimental calibration relative error is within 5% .② The size of the virtual experiment include model 1(101 mm × 200 mm) and the model 2(300 mm × 600 mm) .The model 2 to mesoscopi parameters calibration has scale effect ,but the relative error controlled within 9% .③ There are nonlinear positive relationships between the coefficient of friction of discrete element particles and the angle of internal friction ,and the shear strength ,and the residual strength .When the friction coefficient increased by 0 .1 ,the peak deviator stress average increased 118 .85 kPa and the residual strength average increased 90 .44 kPa .④ The greater the confining pressure ,the weaker the material dilatancy is ,when the confining pressure is changing betw een 100 kPa~500 kPa ,the dilatancy characteristic value K is obtained from 3 to 6 .The cohesive force of the damaged model nearly increases linearly as confining pressure increases .⑤ The greater the Young’s modulus ,the greater the shear strength of the gravel soil is ,but there is not a linear relationship between them .Moreover ,Young’s modulus does not affect residual strength of material significantly .%采用颗粒离散单元方法,实现任意形状块石的模拟,基于室内试验数据标定滑坡坡体物质的堆(残、坡)积碎石土的细观参数,并考虑试样尺

  10. Proline-hydroxylated hypoxia-inducible factor 1α (HIF-1α upregulation in human tumours.

    Cameron E Snell

    Full Text Available The stabilisation of HIF-α is central to the transcriptional response of animals to hypoxia, regulating the expression of hundreds of genes including those involved in angiogenesis, metabolism and metastasis. HIF-α is degraded under normoxic conditions by proline hydroxylation, which allows for recognition and ubiquitination by the von-Hippel-Lindau (VHL E3 ligase complex. The aim of our study was to investigate the posttranslational modification of HIF-1α in tumours, to assess whether there are additional mechanisms besides reduced hydroxylation leading to stability. To this end we optimised antibodies against the proline-hydroxylated forms of HIF-1α for use in formalin fixed paraffin embedded (FFPE immunohistochemistry to assess effects in tumour cells in vivo. We found that HIF-1α proline-hydroxylated at both VHL binding sites (Pro402 and Pro564, was present in hypoxic regions of a wide range of tumours, tumour xenografts and in moderately hypoxic cells in vitro. Staining for hydroxylated HIF-1α can identify a subset of breast cancer patients with poorer prognosis and may be a better marker than total HIF-1α levels. The expression of unhydroxylated HIF-1α positively correlates with VHL in breast cancer suggesting that VHL may be rate-limiting for HIF degradation. Our conclusions are that the degradation of proline-hydroxylated HIF-1α may be rate-limited in tumours and therefore provides new insights into mechanisms of HIF upregulation. Persistence of proline-hydroxylated HIF-1α in perinecrotic areas suggests there is adequate oxygen to support prolyl hydroxylase domain (PHD activity and proline-hydroxylated HIF-1α may be the predominant form associated with the poorer prognosis that higher levels of HIF-1α confer.

  11. Transfer hydrogenation reactions catalyzed by chiral half-sandwich Ruthenium complexes derived from Proline



    Chiral ruthenium half-sandwich complexes were prepared using a chelating diamine made from proline with a phenyl, ethyl, or benzyl group, instead of hydrogen on one of the coordinating arms. Three of these complexes were obtained as single diastereoisomers and their configuration identified by X-ray crystallography. The complexes are recyclable catalysts for the reduction of ketones to chiral alcohols in water. A ruthenium hydride species is identified as the active species by NMR spectroscopy and isotopic labelling experiments.Maximum enantio-selectivity was attained when a phenyl group was directly attached to the primary amine on the diamine ligand derived from proline.

  12. Unconventional N-H…N Hydrogen Bonds Involving Proline Backbone Nitrogen in Protein Structures.

    Deepak, R N V Krishna; Sankararamakrishnan, Ramasubbu


    Contrary to DNA double-helical structures, hydrogen bonds (H-bonds) involving nitrogen as the acceptor are not common in protein structures. We systematically searched N-H…N H-bonds in two different sets of protein structures. Data set I consists of neutron diffraction and ultrahigh-resolution x-ray structures (0.9 Å resolution or better) and the hydrogen atom positions in these structures were determined experimentally. Data set II contains structures determined using x-ray diffraction (resolution ≤ 1.8 Å) and the positions of hydrogen atoms were generated using a computational method. We identified 114 and 14,347 potential N-H…N H-bonds from these two data sets, respectively, and 56-66% of these were of the Ni+1-Hi+1…Ni type, with Ni being the proline backbone nitrogen. To further understand the nature of such unusual contacts, we performed quantum chemical calculations on the model compound N-acetyl-L-proline-N-methylamide (Ace-Pro-NMe) with coordinates taken from the experimentally determined structures. A potential energy profile generated by varying the ψ dihedral angle in Ace-Pro-NMe indicates that the conformation with the N-H…N H-bond is the most stable. An analysis of H-bond-forming proline residues reveals that more than 30% of the proline carbonyl groups are also involved in n → π(∗) interactions with the carbonyl carbon of the preceding residue. Natural bond orbital analyses demonstrate that the strength of N-H…N H-bonds is less than half of that observed for a conventional H-bond. This study clearly establishes the H-bonding capability of proline nitrogen and its prevalence in protein structures. We found many proteins with multiple instances of H-bond-forming prolines. With more than 15% of all proline residues participating in N-H…N H-bonds, we suggest a new, to our knowledge, structural role for proline in providing stability to loops and capping regions of secondary structures in proteins.

  13. Effect of sulphite ions on the proline and polyamine content in the bean Phaseolus vulgaris

    Piotr Karolewski


    Full Text Available The influence of sulphite (0.0125-0.2% on changes in the levels of free proline, protein bound proline and hydroxyproline and of free polyamines in the roots and leaves of two cultivars of beans (Phaseolus vulgaris differing in sensitivity to these ions was investigated. It was found that both cultivars reacted similarily, the response of the seedlings of the more sensitive cultivar occuring at lower concentrations of sulphite.The observed changes in the content of imino acids and polyamines are discussed.

  14. Metabolic network capacity of Escherichia coli for Krebs cycle-dependent proline hydroxylation.

    Theodosiou, Eleni; Frick, Oliver; Bühler, Bruno; Schmid, Andreas


    Understanding the metabolism of the microbial host is essential for the development and optimization of whole-cell based biocatalytic processes, as it dictates production efficiency. This is especially true for redox biocatalysis where metabolically active cells are employed because of the cofactor/cosubstrate regenerative capacity endogenous in the host. Recombinant Escherichia coli was used for overproducing proline-4-hydroxylase (P4H), a dioxygenase catalyzing the hydroxylation of free L-proline into trans-4-hydroxy-L-proline with a-ketoglutarate (a-KG) as cosubstrate. In this whole-cell biocatalyst, central carbon metabolism provides the required cosubstrate a-KG, coupling P4H biocatalytic performance directly to carbon metabolism and metabolic activity. By applying both experimental and computational biology tools, such as metabolic engineering and (13)C-metabolic flux analysis ((13)C-MFA), we investigated and quantitatively described the physiological, metabolic, and bioenergetic response of the whole-cell biocatalyst to the targeted bioconversion and identified possible metabolic bottlenecks for further rational pathway engineering. A proline degradation-deficient E. coli strain was constructed by deleting the putA gene encoding proline dehydrogenase. Whole-cell biotransformations with this mutant strain led not only to quantitative proline hydroxylation but also to a doubling of the specific trans-4-L-hydroxyproline (hyp) formation rate, compared to the wild type. Analysis of carbon flux through central metabolism of the mutant strain revealed that the increased a-KG demand for P4H activity did not enhance the a-KG generating flux, indicating a tightly regulated TCA cycle operation under the conditions studied. In the wild type strain, P4H synthesis and catalysis caused a reduction in biomass yield. Interestingly, the ΔputA strain additionally compensated the associated ATP and NADH loss by reducing maintenance energy demands at comparably low glucose

  15. Transgenic tobacco plants accumulating osmolytes show reduced oxidative damage under freezing stress.

    Parvanova, Daniela; Ivanov, Sergei; Konstantinova, Tatyana; Karanov, Emanuil; Atanassov, Atanas; Tsvetkov, Tsvetan; Alexieva, Vera; Djilianov, Dimitar


    We studied the reaction to the oxidative component of freezing in several tobacco lines, transformed with genes coding for enzymes involved in the synthesis of osmoprotectants (proline, fructan or glycine betaine) along with their wild type. The levels of some oxidative stress markers (leakage of electrolytes, hydrogen peroxide and malondialdehyde) as well as the activity of antioxidative enzymes catalase (EC and guaiacol peroxidase (EC have been followed at acclimation, 12 and 24 h freezing and at recovery. Freezing for 24 h resulted in severe damages for the wild type. A corresponding increase of electrolyte leakage, hydrogen peroxide and malondialdehyde contents, a rise of peroxidase activity and inhibition of catalase activity occurred in the non-transformants. Similar, but significantly lower trend of the same parameters has been found for the transgenic lines. Moreover, the oxidative markers returned to their normal levels when the transformants were able to recover from freezing. It could be speculated that transfer of genes, coding for accumulation of osmoprotectants, is related to reduced intensity of freezing-induced oxidative processes. Our lines and model system could serve as a good prerequisite for additional studies to gain further insights into the complex role of osmoprotectants in freezing tolerance.

  16. Reduced collagen accumulation after major surgery

    Jorgensen, L N; Kallehave, F; Karlsmark, T;


    The preoperative and postoperative wound-healing capacity of 23 patients undergoing elective major abdominal, thoracic or urological surgery was tested objectively by the subcutaneous accumulation of hydroxyproline and proline in an expanded polytetrafluoroethylene (ePTFE) tube. Before scheduled...... surgery two ePTFE tubes were implanted for removal after 5 and 10 days. This was repeated for each patient immediately after surgery. After 10 days a higher amount of hydroxyproline was measured before than after operation (median 2.91 (range 0.37-14.45) versus 1.45 (range 0.26-6.94) micrograms/cm, P = 0.......01)). This decline was significantly higher in the six patients who had a postoperative infection (median 3.02 (range -0.06 to 6.14) versus 0.36 (range -1.56 to 12.60) micrograms/cm, P = 0.02). This study shows that major surgery is associated with impairment of subcutaneous collagen accumulation in a test wound...

  17. Study of enzymatic activity and proline change in potted mini carnation (Dianthus caryophyllus L. flowers under ethylene stress

    M. Karimi


    Full Text Available This study was performed to evaluate the role of ethylene stress, ethylene inhibitors and action treatments on longevity of potted-carnation flowers. For this purpose, first, potted-carnation flowers were pretreated with different concentrations of amino-oxyacetic acid (AOA, benzyladenine (BA and 1-methylcyclopropene (1-MCP. Then, ethephon (as stress treatment was sprayed on the pretreated flowers. Results showed that the lowest longevity of carnation flowers (5 and 6 days, respectively in both studied cultivars (‘Silver pink’ and ‘Lilac on purple’ was related to 30 mg/L ethephon. Maximum flower longevity (11.5 days was related to ‘Lilac on purple’ cultivar. The 0.6 µL/L of 1-MCP treatment, in both cultivars, was the most effective treatment in inhibition of ethephon on reduction of flowers longevity. Also, the lowest amount of ethylene production and the highest enzyme activity was observed when the flowers were pretreated by 0.6 µL/L of 1-MCP before the stress is applied. The highest proline accumulation was related to 30 mg/L ethephon treatment. Maximum α-amylase activity occurred in 30 mg/L ethephon, 10 and 20 mg/L BA and 50 mg/L AOA pretreatments.

  18. Kinetics of absorption of carbon dioxide into aqueous potassium salt of proline

    Paul, Subham; Thomsen, Kaj


    The absorption of carbon dioxide (CO2) into aqueous solution of potassium prolinate (KPr) are studied at 303, 313, and 323K within the salt concentration range of 0.5–3.0kmolm−3 using a wetted wall column absorber. The experimental results are used to interpret the kinetics of the reaction of CO2...

  19. Recyclable L-proline organocatalyst forWieland-Miescher ketone synthesis

    Vivek Srivastava


    Wieland-Miescher ketone 4 was synthesised using L-proline catalyst in the ionic liquid medium via non-selective conjugated addition reaction followed by an enantioselective intermolecular Aldol condensation reaction of triketone 3 intermediate. Short reaction time, recycling of the catalyst, good yield and selectivity are major outcomes of this proposed protocol.

  20. Proline-poor hydrophobic domains modulate the assembly and material properties of polymeric elastin.

    Muiznieks, Lisa D; Reichheld, Sean E; Sitarz, Eva E; Miao, Ming; Keeley, Fred W


    Elastin is a self-assembling extracellular matrix protein that provides elasticity to tissues. For entropic elastomers such as elastin, conformational disorder of the monomer building block, even in the polymeric form, is essential for elastomeric recoil. The highly hydrophobic monomer employs a range of strategies for maintaining disorder and flexibility within hydrophobic domains, particularly involving a minimum compositional threshold of proline and glycine residues. However, the native sequence of hydrophobic elastin domain 30 is uncharacteristically proline-poor and, as an isolated polypeptide, is susceptible to formation of amyloid-like structures comprised of stacked β-sheet. Here we investigated the biophysical and mechanical properties of multiple sets of elastin-like polypeptides designed with different numbers of proline-poor domain 30 from human or rat tropoelastins. We compared the contributions of these proline-poor hydrophobic sequences to self-assembly through characterization of phase separation, and to the tensile properties of cross-linked, polymeric materials. We demonstrate that length of hydrophobic domains and propensity to form β-structure, both affecting polypeptide chain flexibility and cross-link density, play key roles in modulating elastin mechanical properties. This study advances the understanding of elastin sequence-structure-function relationships, and provides new insights that will directly support rational approaches to the design of biomaterials with defined suites of mechanical properties.




    Addition of the Schiff bases derived from reaction of glycine alkyl esters with benzophenoneimine to alpha,beta-unsaturated ketones, followed by hydrogenation of the addition products, leads to 5- or 3,5-substituted prolines. Hydrolysis of the Michael adducts rather than hydrogenation allows

  2. Characteristics and Osmoregulatory Roles of Uptake Systems for Proline and Glycine Betaine in Lactococcus lactis

    Molenaar, Douwe; Hagting, Anja; Alkema, Harmen; Driessen, Arnold J.M.; Konings, Wilhelmus


    Lactococcus lactis subsp. lacti ML3 contains high pools of proline or betaine when grown under conditions of high osmotic strength. These pools are created by specific transport systems. A high-affinity uptake system for glycine betaine (betaine) with a Km of 1.5 µM is expressed constitutively. The

  3. Dimethylsulphopropionate (DMSP) and proline from the surface of the brown alga Fucus vesiculosus inhibit bacterial attachment.

    Saha, M; Rempt, M; Gebser, B; Grueneberg, J; Pohnert, G; Weinberger, F


    It was demonstrated previously that polar and non-polar surface extracts of the brown alga Fucus vesiculosus collected during winter from the Kiel Bight (Germany) inhibited bacterial attachment at natural concentrations. The present study describes the bioassay-guided identification of the active metabolites from the polar fraction. Chromatographic separation on a size-exclusion liquid chromatography column and bioassays identified an active fraction that was further investigated using nuclear magnetic resonance spectroscopy and mass spectrometry. This fraction contained the metabolites dimethylsulphopropionate (DMSP), proline and alanine. DMSP and proline caused the anti-attachment activity. The metabolites were further quantified on the algal surface together with its associated boundary layer. DMSP and proline were detected in the range 0.12-1.08 ng cm(-2) and 0.09-0.59 ng cm(-2), respectively. These metabolites were tested in the concentration range from 0.1 to 1000 ng cm(-2) against the attachment of five bacterial strains isolated from algae and sediment co-occurring with F. vesiculosus. The surface concentrations for 50% inhibition of attachment of these strains were always vesiculosus were also tested, but proved to be the least sensitive. This study shows that DMSP and proline have an ecologically relevant role as surface inhibitors against bacterial attachment on F. vesiculosus.




    Addition of the Schiff bases derived from reaction of glycine alkyl esters with benzophenoneimine to alpha,beta-unsaturated ketones, followed by hydrogenation of the addition products, leads to 5- or 3,5-substituted prolines. Hydrolysis of the Michael adducts rather than hydrogenation allows synthes

  5. Proline Catalyzed Asymmetric Aldol Reaction between Methyl Ketones and 1-Aryl-2,2,2-trifluoroethanones

    QIU Li-Hua; SHEN Zong-Xuan; SHI Chang-Qing; LIU Yan-Hua; ZHANG Ya-Wen


    Direct asymmetric aldol addition of methyl ketones to 2,2,2-trifluoro-1-phenylethanone and its ring-substituted derivatives was achieved using L-proline as a chiral promoter. Various optically active β-trifluoromethyl-β-hydroxy ketones were obtained in almost quantitative yields with moderate enantioselectivities up to 64 % ee.

  6. Proline-rich polypeptides in Alzheimer's disease and neurodegenerative disorders - Therapeutic potential or a mirage?

    Gladkevich, A.; Bosker, F.; Korf, J.; Yenkoyan, K.; Vahradyan, H.; Aghajanov, M.


    The development of effective and safe drugs for a growing Alzheimer disease population is an increasing need at present. Both experimental and clinical evidence support a beneficial effect of proline-rich polypeptides in a number of neurodegenerative diseases, including Alzheimer disease. Experiment

  7. Effect of drought stress on yield, proline and chlorophyll contents in three chickpea cultivars

    Mafakheri, A.; Siosemardeh, A.; Bahramnejad, B.; Struik, P.C.; Sohrabi, Y.


    Drought stress is one of the major abiotic stresses in agriculture worldwide. This study was carried out to investigate the effect of drought stress on proline content, chlorophyll content, photosynthesis and transpiration, stomatal conductance and yield characteristics in three varieties of chickpe

  8. Characteristics and Osmoregulatory Roles of Uptake Systems for Proline and Glycine Betaine in Lactococcus lactis

    Molenaar, Douwe; Hagting, Anja; Alkema, Harmen; Driessen, Arnold J.M.; Konings, Wilhelmus

    Lactococcus lactis subsp. lacti ML3 contains high pools of proline or betaine when grown under conditions of high osmotic strength. These pools are created by specific transport systems. A high-affinity uptake system for glycine betaine (betaine) with a Km of 1.5 µM is expressed constitutively. The

  9. Facial triad modelling using ferrous pyridinyl prolinate complexes : synthesis and catalytic applications

    Moelands, Marcel A. H.; Schamhart, Daniel J.; Folkertsma, Emma; Lutz, Martin; Spek, Anthony L.; Klein Gebbink, Bert


    A series of new chiral pyridinyl prolinate (RPyProR) ligands and their corresponding Fe(II) triflate and chloride complexes are reported. The ligands possess an NN'O coordination motif, as found in the active site of non-heme iron enzymes with the so-called 2-His-1-carboxylate facial triad. The coor

  10. Staphylococcal PknB as the First Prokaryotic Representative of the Proline-Directed Kinases

    Miller, Malgorzata; Donat, Stefanie; Rakette, Sonja; Stehle, Thilo; Kouwen, Thijs R. H. M.; Diks, Sander H.; Dreisbach, Annette; Reilman, Ewoud; Gronau, Katrin; Becher, Doerte; Peppelenbosch, Maikel P.; van Dijl, Jan Maarten; Ohlsen, Knut


    In eukaryotic cell types, virtually all cellular processes are under control of proline-directed kinases and especially MAP kinases. Serine/threonine kinases in general were originally considered as a eukaryote-specific enzyme family. However, recent studies have revealed that orthologues of eukaryo

  11. Amino Acid Salts for Carbon Dioxide Capture: Evaluating l-Prolinate at Desorber Conditions

    Majchrowicz, Magdalena Elzbieta; Brilman, Wim


    Vapor–liquid equilibrium (VLE) measurements for unloaded and CO2-saturated aqueous potassium salt solutions of l-proline have been performed using a high-pressure equilibrium cell. The solubility of CO2 has been determined at temperatures of 363, 393, and 403 K in 3 and 4 mol dm–3 aqueous solutions.

  12. Proline sulphonamide-catalysed Yamada-Otani condensation: reaction development, substrate scope and scaffold reactivity.

    Yang, Hua; Banerjee, Somdev; Carter, Rich G


    The development of a proline sulphonamide-catalysed method for enantioselective and diastereoselective construction of functionalized cyclohexenones is described. Impact of catalyst structure as well as solvent effects and additives are explored. A significant substrate scope is demonstrated by variation of both the aldehyde and the enone components. Diastereoselective derivatization of the cyclohexenone scaffold illustrates its utility as a building block for chemical synthesis.

  13. L-Proline Catalyzed Enamination of β-Dicarbonyl Compounds under Solvent-free Conditions

    KUNDU Dhiman; MAJEE Adinath; HAJRA Alakananda


    A simple,general and efficient method has been developed for synthesis of various β-enamino ketones and esters by reacting β-dicarbonyl compounds with amines using a catalytic amount of L-proline at room temperature under solvent-free conditions in excellent yields.

  14. A Facile Preparation of New Chiral Diamines/Amino-acids (Alcohols) Derived from L-Proline

    FU,Yu-Qin; TAO,Jing-Chao; DING,Li-Na; LIU,Wei-Min


    @@ Living systems use enzymes to catalyze stereoselective reactions with very high fidelity. Since proline was demonstrated as the simplest "enzyme" that mimics type Ⅰ aldolase to catalyze aldol reaction,[1~3] it has attracted much attention in chemistry and other relative fields to design and synthesize highly efficient chemzyme or organocatalysts that can activate normally unreactive substances to useful asymmetric catalytic transformations.

  15. Role of apoptosis-inducing factor, proline dehydrogenase, and NADPH oxidase in apoptosis and oxidative stress

    Becker DF


    Full Text Available Sathish Kumar Natarajan, Donald F BeckerDepartment of Biochemistry and Redox Biology Center, University of Nebraska-Lincoln, Lincoln, NEAbstract: Flavoproteins catalyze a variety of reactions utilizing flavin mononucleotide or flavin adenine dinucleotide as cofactors. The oxidoreductase properties of flavoenzymes implicate them in redox homeostasis, oxidative stress, and various cellular processes, including programmed cell death. Here we explore three critical flavoproteins involved in apoptosis and redox signaling, ie, apoptosis-inducing factor (AIF, proline dehydrogenase, and NADPH oxidase. These proteins have diverse biochemical functions and influence apoptotic signaling by unique mechanisms. The role of AIF in apoptotic signaling is two-fold, with AIF changing intracellular location from the inner mitochondrial membrane space to the nucleus upon exposure of cells to apoptotic stimuli. In the mitochondria, AIF enhances mitochondrial bioenergetics and complex I activity/assembly to help maintain proper cellular redox homeostasis. After translocating to the nucleus, AIF forms a chromatin degrading complex with other proteins, such as cyclophilin A. AIF translocation from the mitochondria to the nucleus is triggered by oxidative stress, implicating AIF as a mitochondrial redox sensor. Proline dehydrogenase is a membrane-associated flavoenzyme in the mitochondrion that catalyzes the rate-limiting step of proline oxidation. Upregulation of proline dehydrogenase by the tumor suppressor, p53, leads to enhanced mitochondrial reactive oxygen species that induce the intrinsic apoptotic pathway. NADPH oxidases are a group of enzymes that generate reactive oxygen species for oxidative stress and signaling purposes. Upon activation, NADPH oxidase 2 generates a burst of superoxide in neutrophils that leads to killing of microbes during phagocytosis. NADPH oxidases also participate in redox signaling that involves hydrogen peroxide-mediated activation of

  16. Role of proline and pyrroline-5-carboxylate metabolism in plant defense against invading pathogens

    Aarzoo eQamar


    Full Text Available Pyrroline-5-carboxylate (P5C is an intermediate product of both proline biosynthesis and catabolism. Recent evidences indicate that proline-P5C metabolism is tightly regulated in plants, especially during pathogen infection and abiotic stress. However, role of P5C and its metabolism in plants has not yet been fully understood. Studies indicate that P5C synthesized in mitochondria has a role in both resistance (R-gene-mediated and nonhost resistance against invading pathogens. Proline dehydrogenase (ProDH and delta-ornithine amino transferase (δOAT-encoding genes, both involved in P5C synthesis in mitochondria are implicated in defense response of Nicotiana benthamiana and Arabidopsis thaliana against bacterial pathogens. Such defense response is proposed to involve salicylic acid-dependent pathway, reactive oxygen species (ROS and hypersensitive response (HR-associated cell death. Recently HR, a form of programmed cell death (PCD, has been proposed to be induced by changes in mitochondrial P5C synthesis or the increase in P5C levels per se in plants inoculated with either a host pathogen carrying suitable avirulent (Avr gene or a nonhost pathogen. Consistently, A. thaliana mutant plants deficient in P5C catabolism showed HR like cell death when grown in external P5C or proline supplemented medium. Similarly, yeast and plant cells under oxidative stress were shown to increase ROS production and PCD due to increase in P5C levels. Similar mechanism has also been reported as one of the triggers for apoptosis in mammalian cells. This review critically analyzes results from various studies and enumerates the pathways for regulation of P5C levels in the plant cell, especially in mitochondria, during pathogen infection. Further, mechanisms regulating P5C- mediated defense responses, namely HR are outlined. This review also provides new insights into the differential role of proline-P5C metabolism in plants exposed to pathogen infection.

  17. S-Nitrosylation of proline-rich tyrosine kinase 2 involves its activation induced by oxygen-glucose deprivation.

    Yan, Xian-Liang; Liu, Dong-Hai; Zhang, Gong-Liang; Hu, Shu-Qun; Chen, Yu-Guo; Xu, Tie


    Previous studies have demonstrated that activation of proline-rich tyrosine kinase 2 (PYK2) in cerebral ischemia is involved in the modulation of N-methyl-d-aspartate-type (NMDA) glutamate receptor activity and Ca(2+) dynamics, resulting in ischemic neuron death ultimately. A number of reports indicate that PYK2 is a redox sensitive kinase that must be activated by an estrogen-induced reactive oxygen species (ROS). However, the mechanism of PYK2 activation remains incompletely illustrated. Accumulating attention is focused on nitric oxide (NO, a free radical) which plays a critical role in cellular signal transduction through stimulus-coupled S-nitrosylation of cysteine residues. Here we reported that PYK2 over-expressed in human embryonic kidney (HEK293) cells was S-nitrosylated (forming SNO-PYK2) by reacting with GSNO, an exogenous NO donor, at one critical cysteine residue (Cys534) with a biotin switch assay. Moreover, our results showed that S-nitrosylation and phosphorylation of PYK2 over-expressed in SH-SY5Y cells was significantly increased after oxygen-glucose deprivation (OGD). We further investigated whether the activation (phosphorylation) of PYK2 was associated with S-nitrosylation following SH-SY5Y cells OGD. Our results showed that the cysteine534 residue (site of S-nitrosylation) mutant PYK2 over-expressed in SH-SY5Y cells diminished S-nitrosylation of PYK2 and inhibited its phosphorylation induced by OGD. In addition, overexpression of the mutant PYK2 protein could prevent nuclear accumulation and abrogate neuronal cell death compared to wild type PYK2 in SH-SY5Y cells induced by OGD. These data suggest that the activation of PYK2 following OGD may be modulated by S-nitrosylation, which provides a new avenue for stroke therapy by targeting the post-translational modification machinery.




    Love, Samuel H. (Bowman Gray School of Medicine, Winston-Salem, N.C.), and Frank H. Hulcher. Green fluorescent pigment accumulated by a mutant of Cellvibrio gilvus. J. Bacteriol. 87:39-45. 1964.-A mutant of Cellvibrio gilvus, designated strain 139A, liberated a green, fluorescent pigment into the surrounding culture medium. A study of the factors which affected the accumulation of this pigment led to the development of a chemically defined medium which supported maximal pigment accumulation in aerated, liquid cultures. d-Glucose, glycine or l-serine, l-phenylalanine, l-proline, and l-lysine comprised the organic components of this medium. The visible absorption spectrum of the pigment showed a maximal band at 400 mmu (pH 7.0). A difference spectrum between reduced and oxidized pigment showed loss of the band at 400 mmu upon oxidation. However, a methanol-extractable, flavinelike compound occurred in the wild strain but not in the mutant. Ferric ions added to the defined medium stimulated growth, with a concomitant reduction of pigment accumulation. Pigment was formed at a maximal rate during the stationary growth phase, and the highest yield was obtained by 18 hr. Organic solvents did not extract the pigment from water solutions. One and sometimes two, compounds absorbing at 400 mmu could be eluted by ion-exchange chromatography on Cellex-P (H(+)), which was used to separate the pigment from other components in the culture supernatants so that the radioactivity of the pigment could be measured. The mutant synthesized C(14)-labeled pigment from d-glucose-U-C(14) and from each of four amino acids (glycine-1-C(14), l-phenylalanine-U-C(14), l-proline-U-C(14), and l-lysine-U-C(14). Delta-Amino-levulenic acid-4-C(14) did not contribute C(14) to the pigment.

  19. Sorption isotherms, kinetic and optimization process of amino acid proline based polymer nanocomposite for the removal of selected textile dyes from industrial wastewater.

    Raghunath, Sharista; Anand, K; Gengan, R M; Nayunigari, Mithil Kumar; Maity, Arjun


    In this article, adsorption and kinetic studies were carried out on three textile dyes, namely Reactive Blue 222 (RB 222), Reactive Red 195 (RR 195) and Reactive Yellow 145 (RY 145). The dyes studied in a mixture were adsorbed under various conditions onto PRO-BEN, a bentonite modified with a new cationic proline polymer (l-proline-epichlorohydrin polymer). The proline polymer was characterized by (1)H NMR, Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS) and TEM. The PRO-BEN composite was characterized by FT-IR, dynamic light scattering (DLS) (zeta potential), TEM imaging, SEM/EDX and X-ray photoelectron spectroscopy (characterize the binding energy). During adsorption studies, factors involving pH, temperature, the initial concentrations of the dyes and the quantity of PRO-BEN used during adsorption were established. The results revealed that the adsorption mechanism was categorized by the Langmuir type 1 isotherm. The adsorption data followed the pseudo-second order kinetic model. The intraparticle diffusion model indicated that adsorption did not only depend on the intraparticle diffusion of the dyes. The thermodynamic parameters verified that the adsorption process was spontaneous and exothermic. The Gibbs free energy values indicated that physisorption had occurred. Successful adsorption of dyes from an industrial effluent was achieved. Desorption studies concluded that PRO-BEN desorbed the dyes better than alumina. This can thereby be viewed as a recyclable remediation material. The PRO-BEN composite could be a cost efficient alternative towards the removal of organic dyes in wastewater treatment. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Inflammatory effects of blood leukocytes: association with vascular function in neuropeptide Y proline 7-genotyped type 2 diabetes patients.

    Kakko, Tanja; Jaakkola, Ulriikka; Raitakari, Olli T; Kallio, Jaana


    We investigated the associations of inflammatory blood cell activation with vascular parameters in patients with type 2 diabetes to elucidate the possible mechanisms of accelerated atherosclerosis observed in subjects with the Leucine 7 to Proline 7 polymorphism (Leu7Pro) in the neuropeptide Y (NPY). Our study included 31 Caucasian patients with type 2 diabetes; 12 of them had the Leu7Pro7 (heterozygous), and 19 had the Leu7Leu7 (wild type) genotype. Vascular parameters were determined by ultrasound methods. Leukocyte analyses were performed from blood samples using flow cytometry. NPY concentrations were determined in plasma. The amount of platelet-granulocyte complexes was positively correlated with NPY concentration (p=0.008) and carotid intima-media thickness (p=0.035) in the Leu7Pro7 group. Interferon gamma (IFN-γ) expression in monocytes correlated negatively with brachial artery flow-mediated dilatation also in the Leu7Pro7 group (p=0.037). The expression of tissue factor on monocytes correlated negatively with brachial artery diameter in the Leu7Pro7 patients as well (p=0.019). The results indicate significant associations between inflammatory cell activation in blood and vascular atherosclerosis in genetically prone subjects, and provide possible mechanistic information about the role of NPY and the Leu7Pro polymorphism in the development of atherosclerosis.

  1. Variance and covariance of accumulated displacement estimates.

    Bayer, Matthew; Hall, Timothy J


    Tracking large deformations in tissue using ultrasound can enable the reconstruction of nonlinear elastic parameters, but poses a challenge to displacement estimation algorithms. Such large deformations have to be broken up into steps, each of which contributes an estimation error to the final accumulated displacement map. The work reported here measured the error variance for single-step and accumulated displacement estimates using one-dimensional numerical simulations of ultrasound echo signals, subjected to tissue strain and electronic noise. The covariance between accumulation steps was also computed. These simulations show that errors due to electronic noise are negatively correlated between steps, and therefore accumulate slowly, whereas errors due to tissue deformation are positively correlated and accumulate quickly. For reasonably low electronic noise levels, the error variance in the accumulated displacement estimates is remarkably constant as a function of step size, but increases with the length of the tracking kernel.

  2. Proline Scan of the hERG Channel S6 Helix Reveals the Location of the Intracellular Pore Gate

    Thouta, Samrat; Sokolov, Stanislav; Abe, Yuki; Clark, Sheldon J.; Cheng, Yen M.; Claydon, Tom W.


    In Shaker-like channels, the activation gate is formed at the bundle crossing by the convergence of the inner S6 helices near a conserved proline-valine-proline motif, which introduces a kink that allows for electromechanical coupling with voltage sensor motions via the S4-S5 linker. Human ether-a-go-go-related gene (hERG) channels lack the proline-valine-proline motif and the location of the intracellular pore gate and how it is coupled to S4 movement is less clear. Here, we show that proline substitutions within the S6 of hERG perturbed pore gate closure, trapping channels in the open state. Performing a proline scan of the inner S6 helix, from Ile655 to Tyr667 revealed that gate perturbation occurred with proximal (I655P-Q664P), but not distal (R665P-Y667P) substitutions, suggesting that Gln664 marks the position of the intracellular gate in hERG channels. Using voltage-clamp fluorimetry and gating current analysis, we demonstrate that proline substitutions trap the activation gate open by disrupting the coupling between the voltage-sensing unit and the pore of the channel. We characterize voltage sensor movement in one such trapped-open mutant channel and demonstrate the kinetics of what we interpret to be intrinsic hERG voltage sensor movement. PMID:24606930

  3. The Antiproton Accumulator (AA)


    A section of the AA where the dispersion (and hence the horizontal beam size) is large. One can distinguish (left to right): A large vacuum-tank, a quadrupole (QDN09*), a bending magnet (BST08), another vacuum-tank, a wide quadrupole (QFW08) and (in the background) a further bending magnet (BST08). The tanks are covered with heating tape for bake-out. The tank left of QDN09 contained the kickers for stochastic pre-cooling (see 790621, 8002234, 8002637X), the other one served mainly as vacuum chamber in the region where the beam was large. Peter Zettwoch works on QFW08. * see: H. Koziol, Antiproton Accumulator Parameter List, PS/AA/Note 84-2 (1984) See under 7911303, 7911597X, 8004261 and 8202324. For photos of the AA in different phases of completion (between 1979 and 1982) see: 7911303, 7911597X, 8004261, 8004608X, 8005563X, 8005565X, 8006716X, 8006722X, 8010939X, 8010941X, 8202324, 8202658X, 8203628X .

  4. Structural characterization of the HIV-1 Vpr N terminus: evidence of cis/trans-proline isomerism.

    Bruns, Karsten; Fossen, Torgils; Wray, Victor; Henklein, Peter; Tessmer, Uwe; Schubert, Ulrich


    The 96-residue human immunodeficiency virus (HIV) accessory protein Vpr serves manifold functions in the retroviral life cycle including augmentation of viral replication in non-dividing host cells, induction of G2 cell cycle arrest, and modulation of HIV-induced apoptosis. Using a combination of dynamic light scattering, circular dichroism, and NMR spectroscopy the N terminus of Vpr is shown to be a unique domain of the molecule that behaves differently from the C-terminal domain in terms of self-association and secondary structure folding. Interestingly, the four highly conserved proline residues in the N terminus are predicted to have a high propensity for cis/trans isomerism. Thus the high resolution structure and folding of a synthetic N-terminal peptide (Vpr1-40) and smaller fragments thereof have been investigated. 1H NMR data indicate Vpr1-40 possesses helical structure between residues 17-32, and for the first time, this helix, which is bound by proline residues, was observed even in aqueous solution devoid of any detergent supplements. In addition, NMR data revealed that all of the proline residues undergo a cis/ trans isomerism to such an extent that approximately 40% of all Vpr molecules possess at least one proline in a cis conformation. This phenomenon of cis/trans isomerism, which is unprecedented for HIV-1 Vpr, not only provides an explanation for the molecular heterogeneity observed in the full-length molecule but also indicates that in vivo the folding and function of Vpr should depend on a cis/trans-proline isomerase activity, particularly as two of the proline residues in positions 14 and 35 show considerable amounts of cis isomers. This prediction correlates well with our recent observation (Zander, K., Sherman, M. P., Tessmer, U., Bruns, K., Wray, V., Prechtel, A. T., Schubert, E., Henklein, P., Luban, J., Neidleman, J., Greene, W. C., and Schubert, U. (2003) J. Biol. Chem. 278, 43170-43181) of a functional interaction between the major

  5. [Effects of drought stress and nitrogen fertilization rate on the accumulation of osmolytes in Jatropha curcas seedlings].

    Yin, Li; Liu, Yong-An; Xie, Cai-Yong; Jiang, Xue; Wang, Yong-Jie; Li, Yin-Hua; Yan, Zhen; Hu, Ting-Xing


    A pot experiment with controlled water supply was conducted to study the effects of different drought stress degree (80% FC, 60% FC, 40% FC, and 20% FC) and nitrogen fertilization rate (0 g x pot(-1), 1.2 g x pot(-1), 3.6 g x pot(-1), and 6.0 g x pot(-1)) on the accumulation of osmolytes in different organs of Jatropha curcas seedlings. Under drought stress, the soluble protei and free proline in seedling shoots and roots and the soluble sugar in seedling shoots had a great accumulation, and the free proline content in seedling leaves had a great increase with increasing drought stress degree. Also under drought stress, the Na+, Ca2+, and Mg2+ all highly accumulated in seedling various organs, while K only accumulated greatly in shoots but slightly in leaves and roots. The effects of nitrogen fertilization on the accumulation of osmolytes in seedlings depended on drought stress degree and nitrogen fertilization rate. At 80% FC and 60% FC, increasing nitrogen fertilization rate could markedly promote the accumulation of osmolytes in the organs of J. curcas seedlings; at 40% FC, applying 6.0 g x pot(-1) weakened the promotion effect on the osmolytes accumulation; whereas at 20%, applying 1.2 g x pot(-1) made the plants have a higher capability in osmoregulation, but applying 3.6 g x pot(-1) and 6.0 g x pot(-1) had less promotion effect, and even, inhibited osmolytes accumulation.

  6. N-Butoxycarbonyl-5-oxo-l-proline ethyl ester

    P. Rajalakshmi


    Full Text Available The molecular structure of the title compound, C12H19NO5, may be visualized as made up of two nearly perpendicular planes [dihedral angle = 87.39 (12°] and its crystal structure is a good example of C—H...O interactions assuming significance in optimizing supramolecular aggregation in crystals in a molecule which is severely imbalanced in terms of donors to acceptor atoms. The pyrrolidine ring adopts a (3T2 twist conformation with puckering parameters Q = 0.2630 (4 Å and ϕ = 59 (9°. The crystal structure features R24(10 and R34(26 ring motifs formed by four weak C—H...O interactions, leading to supramolecular sheets lying parallel to the bc plane.

  7. On the influence of low-energy ionizing radiation on the amino acid molecule: proline

    Tamuliene, Jelena; Romanova, Liudmila; Vukstich, Vasyl; Papp, Alexander; Shkurin, Serhiy; Baliulyte, Laura; Snegursky, Alexander


    New data on the electron-impact fragmentation of the amino acid proline molecule are presented as being related to the formation of the ionized products due to the influence of low-energy ionizing radiation on the above molecule. An extensive DFT-theory based on the theoretical approach enabled the main pathways of the proline molecules fragmentation to be elucidated. A series of the produced fragments have been identified. The absolute appearance energies for some of them have been both measured experimentally and calculated theoretically. The data of the experimental studies and theoretical calculations are compared and analyzed. Contribution to the Topical Issue "Low-Energy Interactions related to Atmospheric and Extreme Conditions", edited by S. Ptasinska, M. Smialek-Telega, A. Milosavljevic, B. Sivaraman.

  8. Pin1-Induced Proline Isomerization in Cytosolic p53 Mediates BAX Activation and Apoptosis.

    Follis, Ariele Viacava; Llambi, Fabien; Merritt, Parker; Chipuk, Jerry E; Green, Douglas R; Kriwacki, Richard W


    The cytosolic fraction of the tumor suppressor p53 activates the apoptotic effector protein BAX to trigger apoptosis. Here we report that p53 activates BAX through a mechanism different from that associated with activation by BH3 only proteins (BIM and BID). We observed that cis-trans isomerization of proline 47 (Pro47) within p53, an inherently rare molecular event, was required for BAX activation. The prolyl isomerase Pin1 enhanced p53-dependent BAX activation by catalyzing cis-trans interconversion of p53 Pro47. Our results reveal a signaling mechanism whereby proline cis-trans isomerization in one protein triggers conformational and functional changes in a downstream signaling partner. Activation of BAX through the concerted action of cytosolic p53 and Pin1 may integrate cell stress signals to induce a direct apoptotic response. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Proline dehydrogenase is a positive regulator of cell death in different kingdoms.

    Cecchini, Nicolás M; Monteoliva, Mariela I; Alvarez, María E


    Proline dehydrogenase (ProDH) catalyzes the flavin-dependent oxidation of Pro into Δ1-pyrroline-5-carboxylate (P5C). This is the first of the two enzymatic reactions that convert proline (Pro) into glutamic acid (Glu). The P5C thus produced is non-enzymatically transformed into glutamate semialdehyde (GSA), which acts as a substrate of P5C dehydrogenase (P5CDH) to generate Glu. Activation of ProDH can generate different effects depending on the behaviour of other enzymes of this metabolism. Under different conditions it can generate toxic levels of P5C, alter the cellular redox homeostasis and even produce reactive oxygen species (ROS). Recent studies indicate that in Arabidopsis, the enzyme potentiates the oxidative burst and cell death associated to the Hypersensitive Responses (HR). Interestingly, activation of ProDH can also produce harmful effects in other organisms, suggesting that the enzyme may play a conserved role in the control of cell death.

  10. Palladium-catalyzed asymmetric Heck arylation of 2,3-dihydrofuran--effect of prolinate salts.

    Morel, Adam; Silarska, Ewelina; Trzeciak, Anna M; Pernak, Juliusz


    Chiral ionic liquids (CILs) containing L-prolinate and L-lactate anions and non-chiral quaternary ammonium cations were employed in the palladium catalyzed enantioselective Heck arylation of 2,3-dihydrofuran with aryl iodides (iodobenzene, 4-iodotoluene, 2-iodoanisole, 4-iodoanisole, 4-iodoacetophenone). In all the reactions 2-aryl-2,3-dihydrofuran (3) was obtained as the main product with the yield up to 52% at the total conversion reaching 83%. Product 3, 2-phenyl-2,3-dihydrofuran, was obtained with excellent enantioselectivity (>99% ee) in a 6 h reaction with tetrabutylammonium L-prolinate. In the proposed homogeneous reaction Pd(0) nanoparticles are considered as a resting state of the catalyst and a source of soluble palladium species catalyzing the Heck reaction. The yield and stereoselectivity of the Heck reaction are strongly influenced by the kind of non-chiral cations present in CILs.

  11. D-Amino acid oxidase and presence of D-proline in Xenopus laevis.

    Soma, Hiroki; Furuya, Ryuji; Kaneko, Ryo; Tsukamoto, Ayaka; Shirasu, Kazumitsu; Tanigawa, Minoru; Nagata, Yoko


    We purified D-amino acid oxidase (EC, DAO) from Xenopus laevis tadpoles. The optimal temperature and pH for enzyme activity were 35-40 °C and 8.3-9.0, respectively, depending on the substrate amino acids available to the enzyme; the highest activity was observed with D-proline followed by D-phenylalanine. Activity was significantly inhibited by p-hydroxymercuribenzoate, but only moderately by p-chloromercuribenzoate or benzoate. Enzyme activity was increased until the final tadpole stage, but was reduced to one-third in the adult and was localized primarily in the kidney. The tadpoles contained high concentrations of D-proline close to the final developmental stage and nearly no D-amino acids were detected in the adult frog, indicating that D-amino acid oxidase functions in metamorphosis.

  12. Specific interactions outside the proline-rich core of two classes of Src homology 3 ligands.

    Feng, S; Kasahara, C; Rickles, R J; Schreiber, S L


    Two dodecapeptides belonging to distinct classes of Src homology 3 (SH3) ligands and selected from biased phage display libraries were used to investigate interactions between a specificity pocket in the Src SH3 domain and ligant residues flanking the proline-rich core. The solution structures of c-Src SH3 complexed with these peptides were solved by NMR. In addition to proline-rich, polyproline type II helix-forming core, the class I and II ligands each possesses a flanking sequence that occupies a large pocket between the RT and n-Src loops of the SH3 domain. Structural and mutational analyses illustrate how the two classes of SH3 ligands exploit a specificity pocket on the receptor differently to increase binding affinity and specificity. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:8618911

  13. Study on Synthesis and Intercalation Behavior of Layered Zirconium Proline-N-methylphosphonate-phosphate

    Xiao Ping RAO; Xiang Kai FU; Kai RAO


    Zirconium proline-N-methylphosphonate-phosphate (α-ZPMPP) was prepared in the presence of HF for the first time. The α-ZPMPP sample is highly crystallized with interlayer distance of 1.52 nm. The interlayer distance of complex of α-ZPMPP with n-butylamine (α-ZPMPP-BA) is in 0.45 nm larger than that of α-ZPMPP. The α-ZPMPP possesses different intercalation behavior of host-guest compound from α-ZP.

  14. Synthesis, docking and anticancer activity studies of D-proline-incorporated wainunuamide

    M Himaja; A Ranjitha; Sunil V Mali


    D-proline-incorporated wainunuamide — a cyclic octapeptide was synthesized and characterized by FTIR, 1H and 13C NMR and Mass spectral analysis. Molecular docking studies were carried out for the designed cyclic octapeptide and the results showed greater affinity for HPV18-2IOI receptor (HeLa cancer cell line). The synthesized cyclic octapeptide exhibited potent anticancer activity against HeLa cancer cells.

  15. Oxidized low-density lipoproteins upregulate proline oxidase to initiate ROS-dependent autophagy

    Zabirnyk, Olga; Liu, Wei; Khalil, Shadi; Sharma, Anit; Phang, James M.


    Epidemiological studies showed that high levels of oxidized low-density lipoproteins (oxLDLs) are associated with increased cancer risk. We examined the direct effect of physiologic concentrations oxLDL on cancer cells. OxLDLs were cytotoxic and activate both apoptosis and autophagy. OxLDLs have ligands for peroxisome proliferator-activated receptor gamma and upregulated proline oxidase (POX) through this nuclear receptor. We identified 7-ketocholesterol (7KC) as a main component responsible ...

  16. Oxidized low-density lipoproteins upregulate proline oxidase to initiate ROS-dependent autophagy


    Epidemiological studies showed that high levels of oxidized low-density lipoproteins (oxLDLs) are associated with increased cancer risk. We examined the direct effect of physiologic concentrations oxLDL on cancer cells. OxLDLs were cytotoxic and activate both apoptosis and autophagy. OxLDLs have ligands for peroxisome proliferator-activated receptor gamma and upregulated proline oxidase (POX) through this nuclear receptor. We identified 7-ketocholesterol (7KC) as a main component responsible ...

  17. Activating Mutations of the TRPML1 Channel Revealed by Proline-scanning Mutagenesis*


    The mucolipin TRP (TRPML) proteins are a family of endolysosomal cation channels with genetically established importance in humans and rodent. Mutations of human TRPML1 cause type IV mucolipidosis, a devastating pediatric neurodegenerative disease. Our recent electrophysiological studies revealed that, although a TRPML1-mediated current can only be recorded in late endosome and lysosome (LEL) using the lysosome patch clamp technique, a proline substitution in TRPML1 (TRPML1V432P) results in a...

  18. Highly Efficient Asymmetric Synthesis Usomg Organocatalyst Derived From(S)-proline



    1 Results Much effort has been focused on organocatalytic asymmetric synthesis in these several years. We have already documented highly efficient organocatalytic asymmetric acylation of a wide variety of racemic alcohols and meso-diols catalyzed bya chiral 1,2-diamine derived from (S)-proline[1]. (S)-Homoproline seems to be a potentially interesting organocatalyst, but no examples using (S)-homoproline itself in asymmetric synthesis has been reported. We have accomplished an efficient and practical syn...

  19. Protected aminooxyprolines for expedited library synthesis: application to Tsg101-directed proline-oxime containing peptides.

    Liu, Fa; Stephen, Andrew G; Fisher, Robert J; Burke, Terrence R


    The stereoselective synthesis of aminooxy-containing proline analogues bearing Fmoc/Boc or Fmoc/Mtt protection that renders them suitable for incorporation into peptides using Fmoc protocols is reported. Acid-catalyzed unmasking at the completion of peptide synthesis yields free aminooxy-functionalities for oxime formation through reaction with libraries of aldehydes. This allows post solid-phase diversification strategies that may facilitate structure-activity relationship studies.

  20. Influence of arbuscular mycorrhizal fungi and copper on growth, accumulation of osmolyte, mineral nutrition and antioxidant enzyme activity of pepper (Capsicum annuum L.).

    Abdel Latef, Arafat Abdel Hamed


    The effect of arbuscular mycorrhizal (AM) fungi inoculation on pepper (Capsicum annuum L. cv. Zhongjiao 105) plant growth and on some physiological parameters in response to increasing soil Cu concentrations was studied. Treatments consisted of inoculation or not with Glomus mosseae and the addition of Cu to soil at the concentrations of 0 (control), 2 (low), 4 (medium), and 8 (high) mM CuSO(4). AM fungal inoculation decreased Cu concentrations in plant organs and promoted biomass yields as well as the contents of chlorophyll, soluble sugar, total protein, and the concentrations of P, K, Ca, and Mg. Plants grown in high Cu concentration exhibited a Cu-induced proline accumulation and also an increase in total free amino acid contents; however, both were lower in mycorrhizal pepper. Cu-induced oxidative stress by increasing lipid peroxidation rates and the activity of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase, and AM symbiosis enhanced these antioxidant enzyme activities and decreased oxidative damage to lipids. In conclusion G. mosseae was able to maintain an efficient symbiosis with pepper plants in contaminated Cu soils, improving plant growth under these conditions, which is likely to be due to reduced Cu accumulation in plant tissues, reduced oxidative stress and damage to lipids, or enhanced antioxidant capacity.

  1. L-proline-based deep eutectic solvents (DESs) for deep catalytic oxidative desulfurization (ODS) of diesel.

    Hao, Lingwan; Wang, Meiri; Shan, Wenjuan; Deng, Changliang; Ren, Wanzhong; Shi, Zhouzhou; Lü, Hongying


    A series of L-proline-based DESs was prepared through an atom economic reaction between L-proline (L-Pro) and four different kinds of organic acids. The DESs were characterized by Fourier transform infrared spectroscopy (FT-IR), H nuclear magnetic resonance ((1)HNMR), cyclic voltammogram (CV) and the Hammett method. The synthesized DESs were used for the oxidative desulfurization and the L-Pro/p-toluenesultonic acid (L-Pro/p-TsOH) system shows the highest catalytic activity that the removal of dibenzothiophene (DBT) reached 99% at 60°C in 2h, which may involve the dual activation of the L-Pro/p-TsOH. The acidity of four different L-proline-based DESs was measured and the results show that it could not simply conclude that the correlation between the acidity of DESs and desulfurization capability was positive or negative. The electrochemical measurements evidences and recycling experiment indicate a good stability performance of L-Pro/p-TsOH in desulfurization. This work will provide a novel and potential method for the deep oxidation desulfurization. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Functional importance of motif I of pseudouridine synthases: mutagenesis of aligned lysine and proline residues.

    Spedaliere, C J; Hamilton, C S; Mueller, E G


    On the basis of sequence alignments, the pseudouridine synthases were grouped into four families that share no statistically significant global sequence similarity, though some common sequence motifs were discovered [Koonin, E. V. (1996) Nucleic Acids. Res. 24, 2411-2415; Gustafsson, C., Reid, R., Greene, P. J., and Santi, D. V. (1996) Nucleic Acids Res. 24, 3756-3762]. We have investigated the functional significance of these alignments by substituting the nearly invariant lysine and proline residues in Motif I of RluA and TruB, pseudouridine synthases belonging to different families. Contrary to our expectations, the altered enzymes display only very mild kinetic impairment. Substitution of the aligned lysine and proline residues does, however, reduce structural stability, consistent with a temperature sensitive phenotype that results from substitution of the cognate proline residue in Cbf5p, a yeast homologue of TruB [Zerbarjadian, Y., King, T., Fournier, M. J., Clarke, L., and Carbon, J. (1999) Mol. Cell. Biol. 19, 7461-7472]. Together, our data support a functional role for Motif I, as predicted by sequence alignments, though the effect of substituting the highly conserved residues was milder than we anticipated. By extrapolation, our findings also support the assignment of pseudouridine synthase function to certain physiologically important eukaryotic proteins that contain Motif I, including the human protein dyskerin, alteration of which leads to the disease dyskeratosis congenita.

  3. δ(15) N from soil to wine in bulk samples and proline.

    Paolini, Mauro; Ziller, Luca; Bertoldi, Daniela; Bontempo, Luana; Larcher, Roberto; Nicolini, Giorgio; Camin, Federica


    The feasibility of using δ(15) N as an additional isotopic marker able to link wine to its area of origin was investigated. The whole production chain (soil-leaves-grape-wine) was considered. Moreover, the research included evaluation of the effect of the fermentation process, the use of different types of yeast and white and red vinification, the addition of nitrogen adjuvants and ultrasound lysis simulating wine ageing. The δ(15) N of grapes and wine was measured in bulk samples and compounds, specifically in proline, for the first time. Despite isotopic fractionation from soil to wine, the δ(15) N values of leaves, grapes, wine and particularly must and wine proline conserved the variability of δ(15) N in the growing soil. Fermentation and ultrasound treatment did not affect the δ(15) N values of grape must, which was therefore conserved in wine. The addition of inorganic or organic adjuvants was able to influence the δ(15) N of bulk wine, depending on the amount and the difference between the δ(15) N of must and that of the adjuvant. The δ(15) N of wine proline was not influenced by adjuvant addition and is therefore the best marker for tracing the geographical origin of wine. Copyright © 2016 John Wiley & Sons, Ltd.

  4. PRGL:A cell wall proline-rich protein containning GASA domain in Gerbera hybrida


    PRPs (proline-rich proteins) are a group of cell wall proteins characterized by their proline and hy- droproline-rich repetitive peptides. The expression of PRPs in plants is stimulated by wounding and environmental stress. GASA (gibberellic acid stimulated in Arabidopsis) proteins are small peptides sharing a 60 amino acid conserved C-terminal domain containing twelve invariant cysteine residues. Most of GASAs reported are localized to apoplasm or cell wall and their expression was regulated by gibberellins (GAs). It has been reported that, in French bean, these two proteins encoding by two distinct genes formed a two-component chitin-receptor involved in plant-pathogen interactions when plant was infected. We cloned a full-length cDNA of PRGL (proline-rich GASA-like) gene which encodes a protein containing both PRP and GASA-like domains. It is demonstrated that PRGL is a new protein with characteristics of PRP and GASA by analyzing its protein structure and gene expression.

  5. Allotides: Proline-Rich Cystine Knot α-Amylase Inhibitors from Allamanda cathartica.

    Nguyen, Phuong Q T; Luu, Thuy T; Bai, Yang; Nguyen, Giang K T; Pervushin, Konstantin; Tam, James P


    Cystine knot α-amylase inhibitors belong to a knottin family of peptidyl inhibitors of 30-32 residues and contain two to four prolines. Thus far, only four members of the group of cystine knot α-amylase inhibitors have been characterized. Herein, the discovery and characterization of five cystine knot α-amylase inhibitors, allotides C1-C5 (Ac1-Ac5) (1-5), from the medicinal plant Allamanda cathartica are reported using both proteomic and genomic methods. Proteomic analysis showed that 1-5 are 30 amino acids in length with three or four proline residues. NMR determination of 4 revealed that it has two cis- and one trans-proline residues and adopts two equally populated conformations in solution. Determination of disulfide connectivity of 2 by differential S-reduction and S-alkylation provided clues of its unfolding process. Genomic analysis showed that allotide precursors contain a three-domain arrangement commonly found in plant cystine knot peptides with conserved residues flanking the processing sites of the mature allotide domain. This work expands the number of known cystine knot α-amylase inhibitors and furthers the understanding of both the structural and biological diversity of this type of knottin family.

  6. Structure and function of the proline-rich region of myelin basic protein.

    Fraser, P E; Deber, C M


    Myelin basic protein (MBP)--the major extrinsic membrane protein of central nervous system myelin--from several species contains a rarely encountered highly conserved triproline segment as residues 99-101 of its 170-residue sequence. Cis peptide bonds are known to arise at X-Pro junctions in proteins and may be of functional significance in protein folding, chain reversal, and/or maintenance of tertiary structure. We have examined the conformation of this proline-rich region using principally 13C nuclear magnetic resonance spectroscopy (125 MHz) both in intact bovine MBP and in several MBP fragment peptides which we synthesized, including octapeptide 97-104 (Arg-Thr-Pro-Pro-Pro-Ser-Gln-Gly). Results suggested an all-trans conformation in aqueous solution for the triproline segment in MBP hexapeptide (99-104), heptapeptide (98-104), and octapeptide. Comparison with the 13C spectrum of intact MBP (125 MHz) suggested that the proline-rich region, as well as all other X-Pro MBP peptide junctures, was also essentially all trans in aqueous solution. Although experiments in which octapeptide 97-104 was bound to a lipid preparation (4:1 dipalmitoylphosphatidylcholine/dimyristoylphosphatidic acid) demonstrated that cis-proline bonds do arise (to the extent of ca. 5%) in the membrane environment, a role of linear chain propagation is suggested for the triproline segment of myelin basic protein.

  7. Long term salinity stress in relation to lipid peroxidation, super oxide dismutase activity and proline content of salt-sensitive and salt-tolerant wheat cultivars Estrés salino a largo plazo en relación con peroxidación lipídica, actividad superóxido dismutasa y contenido de prolina de cultivares de trigo sensibles y tolerantes a la salinidad

    Azam Borzouei


    Full Text Available Salinity is a widespread root medium problem limiting productivity of cereal crops worldwide. The ability of plants to tolerate salt is determined by multiple biochemical pathways that facilitate retention and/or acquisition of water, protect chloroplast functions, and maintain ion homeostasis. Therefore, the ability of salt-sensitive ('Tajan' and salt-tolerant cultivar ('Bam' of Triticum aestivum L. to adapt to a saline environment were evaluated in a set of greenhouse experiments under salt stress during three growth stages (tillering, 50% anthesis, and 10 d after anthesis. Plants were irrigated by different saline waters with electrical conductivities of 1.3, 6, 8, 10, and 12 dS m-1, which were obtained by adding NaCl:CaCl2 in 10:1 molar ratio to fresh water. Differences in growth parameters, lipid peroxidation, superoxide dismutase (SOD activity, and proline accumulation were tested in order to put forward the relative tolerance or sensitivity of cultivars. Results indicated that both parameters differ according to the cultivar's ability in coping oxidative stress caused by salinity. We observed a greater decline in the growth parameters and grain yield under salt stress in 'Tajan' than in 'Bam'. Malondialdehyde content was also higher in 'Tajan'. The improved performance of the 'Bam' under high salinity was accompanied by an increase in SOD (EC activity and proline content at all growth stages. Growth parameters, lipid peroxidation and proline accumulation results are also in good correlation with supporting this cultivar is being relatively tolerant.La salinidad es un problema del medio radical ampliamente distribuido que limita la productividad de los cultivos de cereal en todo el mundo. La capacidad de las plantas para tolerar la sal está determinada por multiples vías bioquímicas que facilitan la retención y/o adquisición de agua, protegen las funciones del cloroplasto, y mantienen la homeostasis iónica. Por lo tanto, se

  8. Tolerance and Accumulation of Heavy Metals by Descurainia sophia L.

    Hoda Karamooz


    Full Text Available Today, biosphere pollution has accelerated strongly with start of industrial revolution by toxicity of heavy metals. One of existing pollution is soil pollution. Unfortunately, soil pollution by metals is as intensive environmental stress for plant hence for human. Plants, which are able to store heavy metals in their organs, can be used for phytoremediation of polluted soils and utilization of these plants is effective for phytoremediation as a cheap and economic method. In this research, the absorption rate of Cd (II, Ni (II by Descurainia sophia was considered in hydroponic conditions. Plants were grown in Hoagland media containing different concentrations of Cd (II, Ni (II. An experiment in a completely randomized design with three replications was conducted. Two weeks after treatment of plants the sample were gathered and metal concentration was measured by atomic absorption spectroscopy. Besides, the content of chlorophyll and proline was measured. The results showed the chlorophyll content in high concentrations of the metals (Cd (II, Ni (II was decreased in plants that were sign of pigment degradation in presence of heavy metals. Similarly, the proline content in plants was increased under stress which was sign of damage of heavy metal stress on plant and activation of defensive mechanisms in this condition. The effects of toxic concentration of nickel and cadmium on metal accumulation in these plants showed that roots were able to absorb more than shoots, which is sign of elements connection to root cell wall.

  9. Enthalpy characteristics of L-proline dissolution in certain water-organic mixtures at 298.15 K

    Badelin, V. G.; Smirnov, V. I.


    A thermochemical study of the processes of L-proline dissolution in aqueous solutions of acetonitrile, 1,4-dioxane, acetone, dimethyl sulfoxide, nitromethane and tetrahydrofuran at T = 298.15 K in the range of organic solvent concentrations x2 = 0-0.25 mole fractions is performed. Standard values of the enthalpies of solution and transfer of L-proline from water to mixed solvent, and the enthalpy coefficients of pairwise interactions between L-proline and molecules of organic solvents, are calculated. The effect the composition of a water-organic mixture and the structure of organic solvents have on the enthalpy characteristics of L-proline dissolution and transfer is examined. The effect the energy properties of intermolecular interactions between components of a mixed solvent has on the intermolecular interactions between L-proline and molecules of cosolvent is estimated. The correlation between the enthalpy characteristics of L-proline dissolution and electron-donor properties of organic cosolvent in aqueous solutions is determined.

  10. The effect of proline on the network structure of major ampullate silks as inferred from their mechanical and optical properties.

    Savage, Ken N; Gosline, John M


    The silk that orb-weaving spiders produce for use as dragline and for the frame of the web is spun from the major ampullate (MA) glands, and it is renowned for its exceptional toughness. The fibroins that make up MA silk have previously been organized into two major groupings, spidroin-1 and spidroin-2, based largely on differences in amino acid sequence. The most apparent difference between spidroin-1 and spidroin-2 fibroins is the lack of proline in spidroin-1. The MA silk of Araneus diadematus comprises two spidroin-2 fibroins, and is therefore proline-rich, whereas spidroin-1 is preferentially expressed in Nephila clavipes MA silk, and so this silk is proline deficient. Together, these two silks provide a system for testing the consequences of proline-rich and proline-deficient fibroin networks. This study measures the mechanical and optical properties of dry and hydrated Araneus and Nephila MA silks. Since proline acts to disrupt secondary structure, it is hypothesized that the fibroin network of Araneus MA silk will contain less secondary structure than the network of Nephila MA silk. Mechanical and optical studies clearly support this hypothesis. Although the dry properties of these two silks are indistinguishable, there are large differences between the hydrated silks. Nephila silk does not swell upon hydration to the same degree as Araneus silk. In addition, upon hydration, Nephila MA silk retains more of its initial dry stiffness, and retains more molecular order, as indicated by birefringence measurements.

  11. Effective prevention of chill-haze in beer using an acid proline-specific endoprotease from Aspergillus niger.

    Lopez, Michel; Edens, Luppo


    Chill-haze formation during beer production is known to involve polyphenols that interact with proline-rich proteins. We hypothesized that incubating beer wort with a proline-specific protease would extensively hydrolyze these proline-rich proteins, yielding a peptide fraction that is unable to form a haze. Predigestion of the proline-rich wheat gliadin with different proteases pointed toward a strong haze-suppressing effect by a proline-specific enzyme. This finding was confirmed in small-scale brewing experiments using a recently identified proline-specific protease with an acidic pH optimum. Subsequent pilot plant trials demonstrated that, upon its addition during the fermentation phase of beer brewing, even low levels of this acidic enzyme effectively prevented chill-haze formation in bottled beer. Results of beer foam stability measurements indicated that the enzyme treatment leaves the beer foam almost unaffected. In combination with the enzyme's cost-effectiveness and regulatory status, these preliminary test results seem to favor further industrial development of this enzymatic beer stabilization method.

  12. Proline catalyzed α-aminoxylation reaction in the synthesis of biologically active compounds.

    Kumar, Pradeep; Dwivedi, Namrata


    The search for new and efficient ways to synthesize optically pure compounds is an active area of research in organic synthesis. Asymmetric catalysis provides a practical, cost-effective, and efficient method to create a variety of complex natural products containing multiple stereocenters. In recent years, chemists have become more interested in using small organic molecules to catalyze organic reactions. As a result, organocatalysis has emerged both as a promising strategy and as an alternative to catalysis with expensive proteins or toxic metals. One of the most successful and widely studied secondary amine-based organocatalysts is proline. This small molecule can catalyze numerous reactions such as the aldol, Mannich, Michael addition, Robinson annulation, Diels-Alder, α-functionalization, α-amination, and α-aminoxylation reactions. Catalytic and enantioselective α-oxygenation of carbonyl compounds is an important reaction to access a variety of useful building blocks for bioactive molecules. Proline catalyzed α-aminoxylation using nitrosobenzene as oxygen source, followed by in situ reduction, gives enantiomerically pure 1,2-diol. This molecule can then undergo a variety of organic reactions. In addition, proline organocatalysis provides access to an assortment of biologically active natural products including mevinoline (a cholesterol lowering drug), tetrahydrolipstatin (an antiobesity drug), R(+)-α-lipoic acid, and bovidic acid. In this Account, we present an iterative organocatalytic approach to synthesize both syn- and anti-1,3-polyols, both enantio- and stereoselectively. This method is primarily based on proline-catalyzed sequential α-aminoxylation and Horner-Wadsworth-Emmons (HWE) olefination of aldehyde to give a γ-hydroxy ester. In addition, we briefly illustrate the broad application of our recently developed strategy for 1,3-polyols, which serve as valuable, enantiopure building blocks for polyketides and other structurally diverse and

  13. Selenium Accumulation and Antioxidant Status of Rice Plants Grown on Seleniferous Soil from Northwestern India

    Sucheta SHARMA


    Full Text Available Greenhouse experiment was conducted to investigate selenium accumulation and its antioxidant response in two rice varieties (PR116 and Pusa Basmati 1121 grown on normal and seleniferous soils. The plant growth was reduced at early developmental stages and flowering was delayed by a period of 10 d on seleniferous soil. Selenium accumulation increased by 3–20 and 13–14 folds in leaves, 18 and 3 folds in grains from Pusa Basmati 1121 and PR116 varieties, respectively. Selenium accumulation in leaves from rice plants grown on seleniferous soil resulted in significant increase in chlorophyll content, hydrogen peroxide, proline, free amino acids, total phenol and tannin contents. Lipid peroxidation levels and peroxidase activities in leaves increased whereas catalase activity showed a reverse trend. It is concluded that selenium accumulation decreased dry matter content in rice during crop development but these plants were able to combat selenium toxicity by inducing alterations in their defense system.

  14. Selenium Accumulation and Antioxidant Status of Rice Plants Grown on Seleniferous Soil from Northwestern India

    Sucheta SHARMA; Reeti GOYAL; Upkar Singh SADANA


    Greenhouse experiment was conducted to investigate selenium accumulation and its antioxidant response in two rice varieties (PR116 and Pusa Basmati 1121) grown on normal and seleniferous soils. The plant growth was reduced at early developmental stages and flowering was delayed by a period of 10 d on seleniferous soil. Selenium accumulation increased by 3–20 and 13–14 folds in leaves, 18 and 3 folds in grains from Pusa Basmati 1121 and PR116 varieties, respectively. Selenium accumulation in leaves from rice plants grown on seleniferous soil resulted in significant increase in chlorophyll content, hydrogen peroxide, proline, free amino acids, total phenol and tannin contents. Lipid peroxidation levels and peroxidase activities in leaves increased whereas catalase activity showed a reverse trend. It is concluded that selenium accumulation decreased dry matter content in rice during crop development but these plants were able to combat selenium toxicity by inducing alterations in their defense system.

  15. Microbial accumulation of uranium, radium, and cesium

    Strandberg, G.W.; Shumate, S.E. II; Parrott, J.R. Jr.; North, S.E.


    Diverse microbial species varied considerably in their ability to accumulate uranium, cesium, and radium. Mechanistic differences in uranium uptake by Saccharomyces cerevisiae and Pseudomonas aeruginosa were indicated. S. serevisiae exhibited a slow (hours) surface accumulation of uranium which was subject to environmental factors, while P. aeruginosa accumulated uranium rapidly (minutes) as dense intracellular deposits and did not appear to be affected by environmental parameters. Metabolism was not required for uranium uptake by either organism. Cesium and radium were concentrated to a considerably lesser extent than uranium by the several species tested.

  16. Identification and characterization of trans-3-hydroxy-l-proline dehydratase and Δ1-pyrroline-2-carboxylate reductase involved in trans-3-hydroxy-l-proline metabolism of bacteria

    Seiya Watanabe


    Full Text Available trans-4-Hydroxy-l-proline (T4LHyp and trans-3-hydroxy-l-proline (T3LHyp occur mainly in collagen. A few bacteria can convert T4LHyp to α-ketoglutarate, and we previously revealed a hypothetical pathway consisting of four enzymes at the molecular level (J Biol Chem (2007 282, 6685–6695; J Biol Chem (2012 287, 32674–32688. Here, we first found that Azospirillum brasilense has the ability to grow not only on T4LHyp but also T3LHyp as a sole carbon source. In A. brasilense cells, T3LHyp dehydratase and NAD(PH-dependent Δ1-pyrroline-2-carboxylate (Pyr2C reductase activities were induced by T3LHyp (and d-proline and d-lysine but not T4LHyp, and no effect of T3LHyp was observed on the expression of T4LHyp metabolizing enzymes: a hypothetical pathway of T3LHyp → Pyr2C → l-proline was proposed. Bacterial T3LHyp dehydratase, encoded to LhpH gene, was homologous with the mammalian enzyme. On the other hand, Pyr2C reductase encoded to LhpI gene was a novel member of ornithine cyclodeaminase/μ-crystallin superfamily, differing from known bacterial protein. Furthermore, the LhpI enzymes of A. brasilense and another bacterium showed several different properties, including substrate and coenzyme specificities. T3LHyp was converted to proline by the purified LhpH and LhpI proteins. Furthermore, disruption of LhpI gene from A. brasilense led to loss of growth on T3LHyp, d-proline and d-lysine, indicating that this gene has dual metabolic functions as a reductase for Pyr2C and Δ1-piperidine-2-carboxylate in these pathways, and that the T3LHyp pathway is not linked to T4LHyp and l-proline metabolism.

  17. Embryogenic callus formation, growth and regeneration in callus and suspension cultures of Miscanthus x ogiformis Honda 'Giganteus' as affected by proline

    Holme, Inger Bæksted; Krogstrup, Peter; Hansen, Jürgen


    The effects of proline additions to culture systems of Miscanthus x ogiformis Honda Giganteus' were investigated. Proline was added in concentrations of 0, 12.5, 25, 50, 100 or 300 mM to the callus induction and suspension culture media containing either Murashige and Skoog or N6 basal salts and 22...... with Murashige and Skoog salts increased embryogenic callus formation on shoot apices and leaf explants while proline had no effect on embryogenic callus formation in medium with N6 salts. Increased growth with increasing proline concentration was obtained in suspension aggregates grown in medium with N6 salts......, whereas proline only increased growth of suspension aggregates grown in medium with Murashige and Skoog salts at concentrations of 12.5 or 25 mM. A stimulating effect of proline on plant regeneration was observed in short-term cultures of callus as well as in long-term cultures of suspension aggregates...

  18. Efeito do estresse salino sobre a concentração de pigmentos e prolina em folhas de arroz Effect of salt stress on pigments and proline concentrations in leaves of rice

    Maria da Graça de Souza Lima


    Full Text Available O experimento foi desenvolvido em casa de vegetação com as cultivares BRS Bojurú, IAS 12-9 Formosa e BRS Agrisul, a fim de analisar o efeito da concentração de NaCl (0, 25, 50, 75 e 100 mM nos teores de clorofilas, carotenóides totais e de prolina, determinados a 30 dias após salinização do solo. A salinidade reduziu fortemente o teor de clorofilas e carotenóides totais no genótipo Agrisul, mas esses pigmentos não foram afetados no Bojurú e Formosa pela concentração salina. O teor de prolina foi maior na cultivar Agrisul do que nas cultivares Bojurú e Formosa e acentuou-se com o incremento na concentração de NaCl. Foi verificada uma concentração salina limiar que inicia o processo de acúmulo de prolina nas folhas, sendo de 50 mM para a cultivar BRS Agrisul (sensível e de 75 mM para BRS Bojurú e IAS 12-9 Formosa (tolerantes.In order to analyse the effect of salt (0, 25, 50, 75 and 100 mM concentration on chlorophylls, total carotenoids and proline contents, an experiment was carried out in greenhouse conditions with cultivars BRS Bojurú, IAS 12-9 Formosa and BRS Agrisul. Total contents were determined 30 days after soil salinization. The salinity strongly reduced chlorophylls and total carotenoids contents on Agrisul genotype, but pigments were not quite affected on Bojuru and Formosa. Proline contents were ligher in Agrisul than in the Bujuru and Formosa genotypes, and the proline accumulation was accentuated with increase in salt concentration. A threshhold of salt concentration that starts the process of proline accumulation in leaves was verified, being 50 mM to cultivar BRS Agrisul (sensitive and 75 mM to cultivars BRS Bojuru and IAS 12-9 Formosa (tolerants.

  19. L-proline increases survival of tilapias infected by Streptococcus agalactiae in higher water temperature.

    Zhao, Xian-Liang; Han, Yi; Ren, Shi-Tong; Ma, Yan-Mei; Li, Hui; Peng, Xuan-Xian


    Streptococcosis causes massive tilapia kills, which results in heavy economic losses of tilapia farming industry. Out of the Streptococcosis, Streptococcus agalactiae is the major pathogen. The bacterium causes higher mortality of tilapias in higher than lower temperatures. However, effect of temperature on metabolic regulation which is related to the mortality is largely unknown. The present study showed 50% and 70% mortality of tilapias cultured in 25 °C and 30 °C, respectively, in comparison with no death in 20 °C following infection caused by S. agalactiae. Then, GC/MS based metabolomics was used to investigate a global metabolic response of tilapia liver to the two higher water temperatures compared to 20 °C. Thirty-six and forty-five varied abundance of metabolites were identified in livers of tilapias cultured at 25 °C and 30 °C, respectively. More decreasing abundance of amino acids and increasing abundance of carbohydrates were detected in 30 °C than 25 °C groups. On the other hand, out of the pathways enriched, the first five biggest impact pathways belong to amino acid metabolism. Decreasing abundance of l-proline was identified as a crucial biomarker for indexing higher water temperature and a potential modulator to reduce the high death. This was validated by engineering injection or oral addition of l-proline. Exogenous l-proline led to elevated amino acid metabolism, which contributes to the elevated survivals. Our findings provide a potential metabolic modulator for controlling the disease, and shed some light on host metabolic prevention to infectious diseases.

  20. A Proline-Based Neuraminidase Inhibitor: DFT Studies on the Zwitterion Conformation, Stability and Formation

    Zhi-Wei Yang


    Full Text Available The designs of potent neuraminidase (NA inhibitors are an efficient way to deal with the recent “2009 H1N1” influenza epidemic. In this work, density functional calculations were employed to study the conformation, stability and formation of the zwitterions of 5-[(1R,2S-1-(acetylamino-2-methoxy-2-methylpentyl]-4-[(1Z-1-propenyl]-(4S,5R-D-proline (BL, a proline-based NA inhibitor. Compared to proline, the zwitterion stability of BL is enhanced by 1.76 kcal mol-1 due to the introduction of functional groups. However, the zwitterion of BL will not represent a local minimum on the potential energy surface until the number of water molecules increases up to two (n = 2. With the addition of two and three water molecules, the energy differences between the zwitterions and corresponding canonical isomers were calculated at 3.13 and -1.54 kcal mol-1, respectively. The zwitterions of BL are mainly stabilized by the H-bonds with the water molecules, especially in the case of three water molecules where the carboxyl-O atoms are largely coordination-saturated by three H-bonds of medium strengths, causing the zwitterion stability even superior to the canonical isomer. With the presence of two and three water molecules, the energy barriers for the conversion processes from the canonical isomers to the zwitterions are equal to 4.96 and 3.13 kcal mol-1, respectively. It indicated that the zwitterion formation is facile to take place with addition of two molecules and further facilitated by more water molecules. Besides, the zwitterion formation of BL is finished in a single step, different from other NA inhibitors. Owing to the above advantages, BL is a good NA inhibitor candidate and more attention should be paid to explorations of BL-based drugs.

  1. Bicyclic proline analogues as organocatalysts for stereoselective aldol reactions: an in silico DFT study.

    Shinisha, C B; Sunoj, Raghavan B


    Density functional theory has been employed in investigating the efficiency of a series of bicyclic analogues of proline as stereoselective organocatalysts for the aldol reaction. Three classes of conformationally restricted proline analogues, as part of either a [2.2.1] or [2.1.1] bicyclic framework, have been studied. Transition states for the stereoselective C-C bond formation between enamines derived from [2.2.1] and [2.1.1] bicyclic amino acids and p-nitrobenzaldehyde, leading to enantiomeric products, have been identified. Analysis of the transition state geometries revealed that the structural rigidity of catalysts, improved transition state organization as well as other weak interactions influence the relative stabilities of diastereomeric transition states and help contribute to the overall stereoselectivity in the aldol reaction. These bicyclic catalysts are predicted to be substantially more effective in improving the enantiomeric excess than the widely used organocatalyst proline. Enantiomeric excesses in the range 82-95% are predicted for these bicyclic catalysts when a sterically unbiased substrate such as p-nitrobenzaldehyde is employed for the asymmetric aldol reaction. More interestingly, introduction of substituents, as simple as a methyl group, at the ortho position of the aryl aldehyde bring about an increase in the enantiomeric excess to values greater than 98%. The reasons behind the vital energy separation between diastereomeric transition states has been rationalized with the help of a number of weak interactions such as intramolecular hydrogen bonding and Coulombic interactions operating on the transition states. These predictions could have wider implications for the rational design of improved organocatalysts for stereoselective carbon-carbon bond-forming reactions.

  2. Characterization of the cell penetrating properties of a human salivary proline-rich peptide.

    Radicioni, Giorgia; Stringaro, Annarita; Molinari, Agnese; Nocca, Giuseppina; Longhi, Renato; Pirolli, Davide; Scarano, Emanuele; Iavarone, Federica; Manconi, Barbara; Cabras, Tiziana; Messana, Irene; Castagnola, Massimo; Vitali, Alberto


    Saliva contains hundreds of small proline-rich peptides most of which derive from the post-translational and post-secretory processing of the acidic and basic salivary proline-rich proteins. Among these peptides we found that a 20 residue proline-rich peptide (p1932), commonly present in human saliva and patented for its antiviral activity, was internalized within cells of the oral mucosa. The cell-penetrating properties of p1932 have been studied in a primary gingival fibroblast cell line and in a squamous cancer cell line, and compared to its retro-inverso form. We observed by mass-spectrometry, flow cytometry and confocal microscopy that both peptides were internalized in the two cell lines on a time scale of minutes, being the natural form more efficient than the retro-inverso one. The cytosolic localization was dependent on the cell type: both peptide forms were able to localize within nuclei of tumoral cells, but not in the nuclei of gingival fibroblasts. The uptake was shown to be dependent on the culture conditions used: peptide internalization was indeed effective in a complete medium than in a serum-free one allowing the hypothesis that the internalization could be dependent on the cell cycle. Both peptides were internalized likely by a lipid raft-mediated endocytosis mechanism as suggested by the reduced uptake in the presence of methyl-ß-cyclodextrin. These results suggest that the natural peptide may play a role within the cells of the oral mucosa after its secretion and subsequent internalization. Furthermore, lack of cytotoxicity of both peptide forms highlights their possible application as novel drug delivery agents.

  3. Physiologically relevant oxidative degradation of oligo(proline) cross-linked polymeric scaffolds.

    Yu, Shann S; Koblin, Rachel L; Zachman, Angela L; Perrien, Daniel S; Hofmeister, Lucas H; Giorgio, Todd D; Sung, Hak-Joon


    Chronic inflammation-mediated oxidative stress is a common mechanism of implant rejection and failure. Therefore, polymer scaffolds that can degrade slowly in response to this environment may provide a viable platform for implant site-specific, sustained release of immunomodulatory agents over a long time period. In this work, proline oligomers of varying lengths (P(n)) were synthesized and exposed to oxidative environments, and their accelerated degradation under oxidative conditions was verified via high performance liquid chromatography and gel permeation chromatography. Next, diblock copolymers of poly(ethylene glycol) (PEG) and poly(ε-caprolactone) (PCL) were carboxylated to form 100 kDa terpolymers of 4%PEG-86%PCL-10%cPCL (cPCL = poly(carboxyl-ε-caprolactone); i% indicates molar ratio). The polymers were then cross-linked with biaminated PEG-P(n)-PEG chains, where P(n) indicates the length of the proline oligomer flanked by PEG chains. Salt-leaching of the polymeric matrices created scaffolds of macroporous and microporous architecture, as observed by scanning electron microscopy. The degradation of scaffolds was accelerated under oxidative conditions, as evidenced by mass loss and differential scanning calorimetry measurements. Immortalized murine bone-marrow-derived macrophages were then seeded on the scaffolds and activated through the addition of γ-interferon and lipopolysaccharide throughout the 9-day study period. This treatment promoted the release of H(2)O(2) by the macrophages and the degradation of proline-containing scaffolds compared to the control scaffolds. The accelerated degradation was evidenced by increased scaffold porosity, as visualized through scanning electron microscopy and X-ray microtomography imaging. The current study provides insight into the development of scaffolds that respond to oxidative environments through gradual degradation for the controlled release of therapeutics targeted to diseases that feature chronic

  4. Alpha-helical, but not beta-sheet, propensity of proline is determined by peptide environment.

    Li, S C; Goto, N K; Williams, K A; Deber, C M


    Proline is established as a potent breaker of both alpha-helical and beta-sheet structures in soluble (globular) proteins. Thus, the frequent occurrence of the Pro residue in the putative transmembrane helices of integral membrane proteins, particularly transport proteins, presents a structural dilemma. We propose that this phenomenon results from the fact that the structural propensity of a given amino acid may be altered to conform to changes imposed by molecular environment. To test this hypothesis on proline, we synthesized model peptides of generic sequence H2N-(Ser-LyS)2-Ala- Leu-Z-Ala-Leu-Z-Trp-Ala-Leu-Z-(Lys-Ser)3-OH (Z = Ala and/or Pro). Peptide conformations were analyzed by circular dichroism spectroscopy in aqueous buffer, SDS, lysophosphatidylglycerol micelles, and organic solvents (methanol, trifluoroethanol, and 2-propanol). The helical propensity of Pro was found to be greatly enhanced in the membrane-mimetic environments of both lipid micelles and organic solvents. Proline was found to stabilize the alpha-helical conformation relative to Ala at elevated temperatures in 2-propanol, an observation that argues against the doctrine that Pro is the most potent alpha-helix breaker as established in aqueous media. Parallel studies in deoxycholate micelles of the temperature-induced conformational transitions of the single-spanning membrane bacteriophage IKe major coat protein, in which the Pro-containing wild type was compared with Pro30 --> Ala mutant, Pro was found to protect the helix, but disrupt the beta-sheet structure as effectively as it does to model peptides in water. The intrinsic capacity of Pro to disrupt beta-sheets was further reflected in a survey of porins where Pro was found to be selectively excluded from the core of membrane-spanning beta-sheet barrels. The overall data provide a rationale for predicting and understanding the structural consequences when Pro occurs in the context of a membrane.

  5. CuI/Proline-catalyzed N-Arylation of Nitrogen Heterocycles


    Ma's CuI/proline procedure for the catalytic cross coupling between nitrogen heterocycles and aryl halides was markedly improved. The key finding was that K3PO4 was a much better base than K2CO3 for the reaction. With this new reaction condition the cross coupling with aryl iodides could be accomplished in 1,4-dioxane instead of DMSO. This reactin also could be carried out in DMF. Furthermore, the coupling yields under the new conditions are usually higher than in Ma's original methods.

  6. DFT Study on the (S)-Proline-catalyzed Direct Aldol Reaction between Acetone and 4-Nitrobenzaldehyde


    DFT/6-31G* calculations were applied to study the direct aldol reaction between acetone and 4-nitrobenzaldehyde catalyzed by (S)-proline. Four transition states associated with the stereo-controlling step, corresponding to syn and anti arrangements of methylene moiety related to the carboxylic acid group in enamine intermediate and re and si attacks to the aldehyde carbonyl carbon have been obtained. The solvent effect of DMSO was investigated with polarized continuum model. The computed energies of transition states reveal the stereo-selectivity of the reaction.

  7. Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus.

    Huihui Cao

    Full Text Available Proline-rich antimicrobial peptides (PR-AMPs are a group of cationic host defense peptides that are characterized by a high content of proline residues. Up to now, they have been reported in some insects, vertebrate and invertebrate animals, but are not found in plants. In this study, we performed an in silico screening of antimicrobial peptides, which led to discovery of a Brassica napus gene encoding a novel PR-AMP. This gene encodes a 35-amino acid peptide with 13 proline residues, designated BnPRP1. BnPRP1 has 40.5% identity with a known proline-rich antimicrobial peptide SP-B from the pig. BnPRP1 was artificially synthetized and cloned into the prokaryotic expression vector pET30a/His-EDDIE-GFP. Recombinant BnPRP1 was produced in Escherichia coli and has a predicted molecular mass of 3.8 kDa. Analysis of its activity demonstrated that BnPRP1 exhibited strong antimicrobial activity against Gram-positive bacterium, Gram-negative bacterium, yeast and also had strong antifungal activity against several pathogenic fungi, such as Sclerotinia sclerotiorum, Mucor sp., Magnaporthe oryzae and Botrytis cinerea. Circular dichroism (CD revealed the main secondary structure of BnPRP1 was the random coil. BnPRP1 gene expression detected by qRT-PCR is responsive to pathogen inoculation. At 48 hours after S. sclerotiorum inoculation, the expression of BnPRP1 increased significantly in the susceptible lines while slight decrease occurred in resistant lines. These suggested that BnPRP1 might play a role in the plant defense response against S. sclerotiorum. BnPRP1 isolated from B. napus was the first PR-AMP member that was characterized in plants, and its homology sequences were found in some other Brassicaceae plants by the genome sequences analysis. Compared with the known PR-AMPs, BnPRP1 has the different primary sequences and antimicrobial activity. Above all, this study gives a chance to cast a new light on further understanding about the AMPs' mechanism

  8. Salicylic acid alleviates adverse effects of heat stress on photosynthesis through changes in proline production and ethylene formation.

    Khan, M Iqbal R; Iqbal, Noushina; Masood, Asim; Per, Tasir S; Khan, Nafees A


    We investigated the potential of salicylic acid (SA) in alleviating the adverse effects of heat stress on photosynthesis in wheat (Triticum aestivum L.) cv WH 711. Activity of ribulose 1,5-bisphosphate carboxylase (Rubisco), photosynthetic-nitrogen use efficiency (NUE), and net photosynthesis decreased in plants subjected to heat stress (40 °C for 6 h), but proline metabolism increased. SA treatment (0.5 mM) alleviated heat stress by increasing proline production through the increase in γ-glutamyl kinase (GK) and decrease in proline oxidase (PROX) activity, resulting in promotion of osmotic potential and water potential necessary for maintaining photosynthetic activity. Together with this, SA treatment restricted the ethylene formation in heat-stressed plants to optimal range by inhibiting activity of 1-aminocyclopropane carboxylic acid (ACC) synthase (ACS). This resulted in improved proline metabolism, N assimilation and photosynthesis. The results suggest that SA interacts with proline metabolism and ethylene formation to alleviate the adverse effects of heat stress on photosynthesis in wheat.

  9. Oral administration of putrescine and proline during the suckling period improves epithelial restitution after early weaning in piglets.

    Wang, J; Li, G R; Tan, B E; Xiong, X; Kong, X F; Xiao, D F; Xu, L W; Wu, M M; Huang, B; Kim, S W; Yin, Y L


    Polyamines are necessary for normal integrity and the restitution after injury of the gastrointestinal epithelium. The objective of this study was to investigate the effects of oral administration of putrescine and proline during the suckling period on epithelial restitution after early weaning in piglets. Eighteen neonatal piglets (Duroc × Landrace × Large Yorkshire) from 3 litters (6 piglets per litter) were assigned to 3 groups, representing oral administration with an equal volume of saline (control), putrescine (5 mg/kg BW), and proline (25 mg/kg BW) twice daily from d 1 to weaning at 14 d of age. Plasma and intestinal samples were obtained 3 d after weaning. The results showed that oral administration of putrescine or proline increased the final BW and ADG of piglets compared with the control (P putrescine- and proline-treated piglets compared with those of control piglets. The voltage-gated K+ channel (Kv) 1.1 protein expression in the jejunum of piglets administrated with putrescine and the Kv1.5 mRNA and Kv1.1 protein levels in the ileum of piglets administrated with proline were greater than those in control piglets (P < 0.05). These findings indicate that polyamine or its precursor could improve mucosal proliferation, intestinal morphology, as well as tight junction and potassium channel protein expressions in early-weaned piglets, with implications for epithelial restitution and barrier function after stress injury.

  10. Intracisternally Injected L-Proline Activates Hypothalamic Supraoptic, but Not Paraventricular, Vasopressin-Expressing Neurons in Conscious Rats

    Yumi Takemoto


    Full Text Available When injected into specific rat brain regions, the neurotransmitter candidate L-proline produces various cardiovascular changes through ionotropic excitatory amino acid receptors. The present study used an immunohistochemical double-labeling approach to determine whether intracisternally injected L-proline in freely moving rats, which increases blood pressure, activates hypothalamic vasopressin-expressing neurons and ventral medullary tyrosine-hydroxylase- (TH- containing neurons. Following injection of L-proline, the number of activated hypothalamic neurons that coexpressed vasopressin and c-Fos was much greater in the supraoptic nucleus (SON than in the paraventricular nucleus (PVN of rats with increased blood pressure. The number of activated TH-containing neurons was significantly greater following L-proline treatment than following control injections of artificial cerebrospinal fluid (ACSF. These results clearly demonstrate that intracisternally injected L-proline activates hypothalamic supraoptic, but not paraventricular, vasopressin-expressing neurons and medullary TH-containing (A1/C1 neurons in freely moving rats.

  11. A Plasmodium falciparum S33 proline aminopeptidase is associated with changes in erythrocyte deformability.

    da Silva, Fabio L; Dixon, Matthew W A; Stack, Colin M; Teuscher, Franka; Taran, Elena; Jones, Malcolm K; Lovas, Erica; Tilley, Leann; Brown, Christopher L; Trenholme, Katharine R; Dalton, John P; Gardiner, Donald L; Skinner-Adams, Tina S


    Infection with the apicomplexan parasite Plasmodium falciparum is a major cause of morbidity and mortality worldwide. One of the striking features of this parasite is its ability to remodel and decrease the deformability of host red blood cells, a process that contributes to disease. To further understand the virulence of Pf we investigated the biochemistry and function of a putative Pf S33 proline aminopeptidase (PfPAP). Unlike other P. falciparum aminopeptidases, PfPAP contains a predicted protein export element that is non-syntenic with other human infecting Plasmodium species. Characterization of PfPAP demonstrated that it is exported into the host red blood cell and that it is a prolyl aminopeptidase with a preference for N-terminal proline substrates. In addition genetic deletion of this exopeptidase was shown to lead to an increase in the deformability of parasite-infected red cells and in reduced adherence to the endothelial cell receptor CD36 under flow conditions. Our studies suggest that PfPAP plays a role in the rigidification and adhesion of infected red blood cells to endothelial surface receptors, a role that may make this protein a novel target for anti-disease interventions strategies.

  12. Dcp1 links coactivators of mRNA decapping to Dcp2 by proline recognition

    Borja, Mark S.; Piotukh, Kirill; Freund, Christian; Gross, John D.


    Cap hydrolysis is a critical step in several eukaryotic mRNA decay pathways and is carried out by the evolutionarily conserved decapping complex containing Dcp2 at the catalytic core. In yeast, Dcp1 is an essential activator of decapping and coactivators such as Edc1 and Edc2 are thought to enhance activity, though their mechanism remains elusive. Using kinetic analysis we show that a crucial function of Dcp1 is to couple the binding of coactivators of decapping to activation of Dcp2. Edc1 and Edc2 bind Dcp1 via its EVH1 proline recognition site and stimulate decapping by 1000-fold, affecting both the KM for mRNA and rate of the catalytic step. The C-terminus of Edc1 is necessary and sufficient to enhance the catalytic step, while the remainder of the protein likely increases mRNA binding to the decapping complex. Lesions in the Dcp1 EVH1 domain or the Edc1 proline-rich sequence are sufficient to block stimulation. These results identify a new role of Dcp1, which is to link the binding of coactivators to substrate recognition and activation of Dcp2. PMID:21148770

  13. A proline racemase based PCR for identification of Trypanosoma vivax in cattle blood.

    Regassa Fikru

    Full Text Available A study was conducted to develop a Trypanosoma vivax (T. vivax specific PCR based on the T. vivax proline racemase (TvPRAC gene. Forward and reverse primers were designed that bind at 764-783 bp and 983-1002 bp of the gene. To assess its specificity, TvPRAC PCR was conducted on DNA extracted from different haemotropic pathogens: T. vivax from Nigeria, Ethiopia and Venezuela, T. congolense Savannah type, T. brucei brucei, T. evansi, T. equiperdum, T. theileri, Theileria parva, Anaplasma marginale, Babesia bovis and Babesia bigemina and from bovine, goat, mouse, camel and human blood. The analytical sensitivity of the TvPRAC PCR was compared with that of the ITS-1 PCR and the 18S PCR-RFLP on a dilution series of T. vivax DNA in water. The diagnostic performance of the three PCRs was compared on 411 Ethiopian bovine blood specimens collected in a former study. TvPRAC PCR proved to be fully specific for T. vivax, irrespective of its geographical origin. Its analytical sensitivity was lower than that of ITS-1 PCR. On these bovine specimens, TvPRAC PCR detected 8.3% T. vivax infections while ITS-1 PCR and 18S PCR-RFLP detected respectively 22.6 and 6.1% T. vivax infections. The study demonstrates that a proline racemase based PCR could be used, preferably in combination with ITS-1 PCR, as a species-specific diagnostic test for T. vivax infections worldwide.

  14. Gene Mining for Proline Based Signaling Proteins in Cell Wall of Arabidopsis thaliana

    Ihsan, Muhammad Z.; Ahmad, Samina J. N.; Shah, Zahid Hussain; Rehman, Hafiz M.; Aslam, Zubair; Ahuja, Ishita; Bones, Atle M.; Ahmad, Jam N.


    The cell wall (CW) as a first line of defense against biotic and abiotic stresses is of primary importance in plant biology. The proteins associated with cell walls play a significant role in determining a plant's sustainability to adverse environmental conditions. In this work, the genes encoding cell wall proteins (CWPs) in Arabidopsis were identified and functionally classified using geneMANIA and GENEVESTIGATOR with published microarrays data. This yielded 1605 genes, out of which 58 genes encoded proline-rich proteins (PRPs) and glycine-rich proteins (GRPs). Here, we have focused on the cellular compartmentalization, biological processes, and molecular functioning of proline-rich CWPs along with their expression at different plant developmental stages. The mined genes were categorized into five classes on the basis of the type of PRPs encoded in the cell wall of Arabidopsis thaliana. We review the domain structure and function of each class of protein, many with respect to the developmental stages of the plant. We have then used networks, hierarchical clustering and correlations to analyze co-expression, co-localization, genetic, and physical interactions and shared protein domains of these PRPs. This has given us further insight into these functionally important CWPs and identified a number of potentially new cell-wall related proteins in A. thaliana. PMID:28289422

  15. Interaction of serum amyloid P component with hexanoyl bis(d-proline) (CPHPC)

    Kolstoe, Simon E. [University College London, Rowland Hill Street, London NW3 2PF (United Kingdom); Jenvey, Michelle C. [University of Southampton, Southampton SO17 1BJ (United Kingdom); Purvis, Alan [Imperial College London, London SW7 2AZ (United Kingdom); Light, Mark E. [University of Southampton, Southampton SO17 1BJ (United Kingdom); Thompson, Darren [University of Sussex, Falmer, Brighton BN1 9RQ (United Kingdom); Hughes, Peter; Pepys, Mark B.; Wood, Stephen P., E-mail: [University College London, Rowland Hill Street, London NW3 2PF (United Kingdom)


    Serum amyloid P component is a pentameric plasma glycoprotein that recognizes and binds to amyloid fibres in a calcium-dependent fashion and is likely to contribute to their deposition and persistence in vivo. Five molecules of the drug CPHPC avidly cross-link pairs of protein pentamers and the decameric complex is rapidly cleared in vivo. Crystal structures of the protein in complex with a bivalent drug and cadmium ions, which improve crystal quality, allow the definition of the preferred bound drug isomers. Under physiological conditions, the pentameric human plasma protein serum amyloid P component (SAP) binds hexanoyl bis(d-proline) (R-1-(6-[R-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl) pyrrolidine-2-carboxylic acid; CPHPC) through its d-proline head groups in a calcium-dependent interaction. Cooperative effects in binding lead to a substantial enhancement of affinity. Five molecules of the bivalent ligand cross-link and stabilize pairs of SAP molecules, forming a decameric complex that is rapidly cleared from the circulation by the liver. Here, it is reported that X-ray analysis of the SAP complex with CPHPC and cadmium ions provides higher resolution detail of the interaction than is observed with calcium ions. Conformational isomers of CPHPC observed in solution by HPLC and by X-ray analysis are compared with the protein-bound form. These are discussed in relation to the development of CPHPC to provide SAP depletion for the treatment of amyloidosis and other indications.

  16. Natural Proline-Rich Cyclopolypeptides from Marine Organisms: Chemistry, Synthetic Methodologies and Biological Status

    Wan-Yin Fang


    Full Text Available Peptides have gained increased interest as therapeutics during recent years. More than 60 peptide drugs have reached the market for the benefit of patients and several hundreds of novel therapeutic peptides are in preclinical and clinical development. The key contributor to this success is the potent and specific, yet safe, mode of action of peptides. Among the wide range of biologically-active peptides, naturally-occurring marine-derived cyclopolypeptides exhibit a broad range of unusual and potent pharmacological activities. Because of their size and complexity, proline-rich cyclic peptides (PRCPs occupy a crucial chemical space in drug discovery that may provide useful scaffolds for modulating more challenging biological targets, such as protein-protein interactions and allosteric binding sites. Diverse pharmacological activities of natural cyclic peptides from marine sponges, tunicates and cyanobacteria have encouraged efforts to develop cyclic peptides with well-known synthetic methods, including solid-phase and solution-phase techniques of peptide synthesis. The present review highlights the natural resources, unique structural features and the most relevant biological properties of proline-rich peptides of marine-origin, focusing on the potential therapeutic role that the PRCPs may play as a promising source of new peptide-based novel drugs.

  17. Phytotoxicity of cadmium on protein, proline and antioxidant enzyme activities in growing Arachis hypogaea L. seedlings


    Phytotoxicity of cadmium on growing Arachis hypogaea L. seedlings was studied. Seeds were exposed to 25, 50, and 100 μmol/L CdCl2 concentrations, for a period of 10, 15, 20 and 25 d. The extent of damage to chlorophyll, protein, proline, nitrate and nitrite reductase, antioxidant enzyme activity in leaves and roots were evaluated after 10 d of cadmium stress. The higher concentration of cadmium (100 μmol/L) resulted (leaves and roots) total chlorophyll 91.01%, protein 79.51%, 83.61%, nitrate reductase 79.39%,80.72% and nitrite reductase 77.07%, 75.88% activity decreased with increase in cadmium concentrations and exposure periods. Cadmium caused significant changes in the activity of antioxidative enzymes. Contrastingly Cd treated plant tissues showed an increase in proline 159.87%, 239.6%, gluthion reductase (GR) 337.72%, 306.14%, superoxide disumutase (SOD) 688.56%, 381.72%, ascorbate peroxidase (APX) 226.47%, 252.14%, peroxidase (POD) 72.19%, 60.29% and catalase (CAT) 228.96%, 214.74% as compared to control. Cadmium stress caused a significant increase in the rate of SOD activity in leaves and roots of plant species. Results show the crop A. hypogaea is highly sensitive even at very low cadmium concentrations.

  18. Relationship between depression anxiety stress scale (DASS) and urinary hydroxyproline and proline concentrations in hospital workers.

    Lee, Keou Won; Kim, Soo Jeong; Park, Jae Beom; Lee, Kyung Jong


    Although increased reactive oxygen species (ROS) is caused by stress accelerates collagen degradation, there was no data on the relationship between stress and urinary hydroxyproline (Hyp) and proline (Pro), a good marker of collagen degradation. The purpose of this study was to evaluate the relationship between depression, anxiety, and stress (DAS) and concentrations of urinary Hyp and Pro. 97 hospital employees aged 20 to 58 were asked to fill out comprehensive self-administrated questionnaires containing information about their medical history, lifestyle, length of the work year, shift-work and DAS. depression anxiety stress scale (DASS) was applied to evaluate chronic mental disorders. Urine samples were analyzed using high performance liquid chromatography (HPLC) with double derivatization for the assay of hydroxyproline and proline. The mean value of Hyp and Pro concentration in all subjects was 194.1 ± 113.4 μmol/g and 568.2 ± 310.7 μmol/g. DASS values and urinary Pro concentrations were differentiated by sex (female > male, p others, p stress (Adjusted r2 = 0.051) and anxiety and job (Adjusted r2 = 0.199), respectively. We found that stress and anxiety were correlated with urinary Hyp and Pro concentrations. To identifying a definite correlation, further study in large populations will be needed.

  19. Proline-arginine rich (PR-39) cathelicidin: Structure, expression and functional implication in intestinal health.

    Holani, Ravi; Shah, Chaitanya; Haji, Qahir; Inglis, G Douglas; Uwiera, Richard R E; Cobo, Eduardo R


    Proline-Arginine-39 (PR-39) is a small cationic, proline and arginine rich, cathelicidin that plays an important role in the porcine innate immune system. Although PR-39 was first discovered in intestinal cell lysates of pigs, subsequent research has indicated that it is primarily expressed in bone marrow and other lymphoid tissues including the thymus and spleen, as well as in leukocytes. Mature PR-39 cathelicidin has anti-microbial activity against many gram-negative and some gram-positive bacteria. PR-39 is also a bridge between the innate and adaptive immune system with recognized immunomodulatory, wound healing, anti-apoptotic, and pro-angiogenic functions. The purpose of this review is to summarize our current knowledge about the structure, expression, and functions of PR-39 and its potential to promote intestinal homeostasis. This understanding is relevant in the search of alternative therapeutics against diarrheic enterocolitis, a major problem faced by pork producers both in terms of costs and risk of zoonosis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Vermicompost humic acids modulate the accumulation and metabolism of ROS in rice plants.

    García, Andrés Calderín; Santos, Leandro Azevedo; de Souza, Luiz Gilberto Ambrósio; Tavares, Orlando Carlos Huertas; Zonta, Everaldo; Gomes, Ernane Tarcisio Martins; García-Mina, José Maria; Berbara, Ricardo Luis Louro


    This work aims to determine the reactive oxygen species (ROS) accumulation, gene expression, anti-oxidant enzyme activity, and derived effects on membrane lipid peroxidation and certain stress markers (proline and malondialdehyde-MDA) in the roots of unstressed and PEG-stressed rice plants associated with vermicompost humic acid (VCHA) application. The results show that the application of VCHA to the roots of unstressed rice plants caused a slight but significant increase in root ROS accumulation and the gene expression and activity of the major anti-oxidant enzymes (superoxide dismutase and peroxidase). This action did not have negative effects on root development, and an increase in both root growth and root proliferation occurred. However, the root proline and MDA concentrations and the root permeability results indicate the development of a type of mild stress associated with VCHA application. When VCHA was applied to PEG-stressed plants, a clear alleviation of the inhibition in root development linked to PEG-mediated osmotic stress was observed. This was associated with a reduction in root ROS production and anti-oxidant enzymatic activity caused by osmotic stress. This alleviation of stress caused by VCHA was also reflected as a reduction in the PEG-mediated concentration of MDA in the root as well as root permeability. In summary, the beneficial action of VCHA on the root development of unstressed or PEG-stressed rice plants clearly involves the modulation of ROS accumulation in roots.

  1. Improving Co-Amorphous Drug Formulations by the Addition of the Highly Water Soluble Amino Acid, Proline

    Katrine Tarp Jensen


    Full Text Available Co-amorphous drug amino acid mixtures were previously shown to be a promising approach to create physically stable amorphous systems with the improved dissolution properties of poorly water-soluble drugs. The aim of this work was to expand the co-amorphous drug amino acid mixture approach by combining the model drug, naproxen (NAP, with an amino acid to physically stabilize the co-amorphous system (tryptophan, TRP, or arginine, ARG and a second highly soluble amino acid (proline, PRO for an additional improvement of the dissolution rate. Co-amorphous drug-amino acid blends were prepared by ball milling and investigated for solid state characteristics, stability and the dissolution rate enhancement of NAP. All co-amorphous mixtures were stable at room temperature and 40 °C for a minimum of 84 days. PRO acted as a stabilizer for the co-amorphous system, including NAP–TRP, through enhancing the molecular interactions in the form of hydrogen bonds between all three components in the mixture. A salt formation between the acidic drug, NAP, and the basic amino acid, ARG, was found in co-amorphous NAP–ARG. In comparison to crystalline NAP, binary NAP–TRP and NAP–ARG, it could be shown that the highly soluble amino acid, PRO, improved the dissolution rate of NAP from the ternary co-amorphous systems in combination with either TRP or ARG. In conclusion, both the solubility of the amino acid and potential interactions between the molecules are critical parameters to consider in the development of co-amorphous formulations.

  2. Dynamical role of phosphorylation on serine/threonine-proline Pin1 substrates from constant force molecular dynamics simulations.

    Velazquez, Hector A; Hamelberg, Donald


    Cis-trans isomerization of peptidyl-prolyl bonds of the protein backbone plays an important role in numerous biological processes. Cis-trans isomerization can be the rate-limiting step due its extremely slow dynamics, compared to the millisecond time scale of many processes, and is catalyzed by a widely studied family of peptidyl-prolyl cis-trans isomerase enzymes. Also, mechanical forces along the peptide chain can speed up the rate of isomerization, resulting in "mechanical catalysis," and have been used to study peptidyl-prolyl cis-trans isomerization and other mechanical properties of proteins. Here, we use constant force molecular dynamics simulations to study the dynamical effects of phosphorylation on serine/threonine-proline protein motifs that are involved in the function of many proteins and have been implicated in many aberrant biological processes. We show that the rate of cis-trans isomerization is slowed down by phosphorylation, in excellent agreement with experiments. We use a well-grounded theory to describe the force dependent rate of isomerization. The calculated rates at zero force are also in excellent agreement with experimentally measured rates, providing additional validation of the models and force field parameters. Our results suggest that the slowdown in the rate upon phosphorylation is mainly due to an increase in the friction along the peptidyl-prolyl bond angle during isomerization. Our results provide a microscopic description of the dynamical effects of post-translational phosphorylation on cis-trans isomerization and insights into the properties of proteins under tension.

  3. Accumulation by Conservation

    Büscher, Bram; Fletcher, Robert


    Following the financial crisis and its aftermath, it is clear that the inherent contradictions of capitalist accumulation have become even more intense and plunged the global economy into unprecedented turmoil and urgency. Governments, business leaders and other elite agents are frantically searchin

  4. Effect of subsoiling in fallow period on soil water storage and grain protein accumulation of dryland wheat and its regulatory effect by nitrogen application.

    Min Sun

    Full Text Available To provide a new way to increase water storage and retention of dryland wheat, a field study was conducted at Wenxi experimental site of Shanxi Agricultural University. The effect of subsoiling in fallow period on soil water storage, accumulation of proline, and formation of grain protein after anthesis were determined. Our results showed that subsoiling in fallow period could increase water storage in the 0-300 cm soil at pre-sowing stage and at anthesis stage with low or medium N application, especially for the 60-160 cm soil. However, the proline content, glutamine synthetase (GS activity, glutamate dehydrogenase (GDH activity in flag leaves and grains were all decreased by subsoiling in fallow period. In addition, the content of albumin, gliadin, and total protein in grains were also decreased while globulin content, Glu/Gli, protein yield, and glutelin content were increased. With N application increasing, water storage of soil layers from 20 to 200 cm was decreased at anthesis stage. High N application resulted in the increment of proline content and GS activity in grains. Besides, correlation analysis showed that soil storage in 40-160 cm soil was negatively correlated with proline content in grains; proline content in grains was positively correlated with GS and GDH activity in flag leaves. Contents of albumin, globulin and total protein in grains were positively correlated with proline content in grains and GDH activity in flag leaves. In conclusion, subsoiling in fallow period, together with N application at 150 kg·hm(-2, was beneficial to increase the protein yield and Glu/Gli in grains which improve the quality of wheat.

  5. Multiple gas-phase conformations of proline-containing peptides: Is it always cis/trans isomerization?

    Lietz, Christopher B.; Chen, Zhengwei; Son, Chang Yun; Pang, Xueqin; Cui, Qiang; Li, Lingjun


    Ion mobility-mass spectrometry (IM-MS) is often employed to look at the secondary, tertiary, and quaternary structure of naked peptides and proteins in the gas-phase. Recently, it has offered a unique glimpse into proline-containing peptides and their cis/trans Xxx-Pro isomers. An experimental “signature” has been identified wherein a proline-containing peptide has its Pro residues substituted with another amino acid and the presence or absence of conformations in the IM-MS spectra are observ...


    Nada Parađiković


    Full Text Available The research of light and substrate role in leaf photosynthetic pigments, free proline content and flower quality indices in two gerbera cultivars (Vino and Ruby Red showed that flower stems were longer in additional lighting treatment, coco’s peat amended with rice chaff and cultivar Ruby Red. Chlorophyll b concentration was in negative relation to leaf proline content and correlated to longer flower stems. In general, the higher leaf total chlorophyll concentration was related to longer stems, and such stems have had larger flower heads. Flower head diameter did not differ significantly between cultivars; it depended on light regime, substrate and their interaction.

  7. Crecimiento y contenido de prolina y carbohidratos de plántulas de frijol sometidas a estrés por sequía Growth and, proline and carbohydrate content of bean seedlings subjected to drought stress

    Teresa Susana Herrera Flores


    Durango race, in order to identify quick features to measure their reliability for selection of resistance to drought stress. The radicle of the seedlings were subjected to four moisture potentials: -2.07, -0.52, -0.27 megapascals (MPa and a control -0.16 MPa. For determining the soluble carbohydrates and proline, the potential -0.16 Mpa was removed, since it caused the hypocotyl rot at the junction with the cotyledon, in this case the potential of-0.27 was considered as control. The radicle of both varieties had greater length in the treatments with less stress. In the most critical potential, -2.07 MPa, Pinto Villa, accumulated more proline and sucrose than Bayo Madero, while starch leveis were quite similar. Highly significant correlation was observed between the elongation of the radiocle with the accumulation of proline, glucose and fructose (r= 0.99 respectively. The accumulation of compatible solutes (glucose, fructose, sucrose and proline in the radicle developed under conditions of stress, favors its growth in early stages of development. Compatible solutes are suitable as biochemical indicators for selection of varieties resistant to moisture stress during the first three days of germination.

  8. Markov models for accumulating mutations

    Beerenwinkel, Niko


    We introduce and analyze a waiting time model for the accumulation of genetic changes. The continuous time conjunctive Bayesian network is defined by a partially ordered set of mutations and by the rate of fixation of each mutation. The partial order encodes constraints on the order in which mutations can fixate in the population, shedding light on the mutational pathways underlying the evolutionary process. We study a censored version of the model and derive equations for an EM algorithm to perform maximum likelihood estimation of the model parameters. We also show how to select the maximum likelihood poset. The model is applied to genetic data from different cancers and from drug resistant HIV samples, indicating implications for diagnosis and treatment.

  9. Novel Potent Hepatitis C Virus NS3 Serine Protease Inhibitors Derived from Proline-Based Macrocycles

    Chen, Kevin X.; Njoroge, F. George; Arasappan, Ashok; Venkatraman, Srikanth; Vibulbhan, Bancha; Yang, Weiying; Parekh, Tejal N.; Pichardo, John; Prongay, Andrew; Cheng, Kuo-Chi; Butkiewicz, Nancy; Yao, Nanhua; Madison, Vincent; Girijavallabhan, Viyyoor (SPRI)


    The hepatitis C virus (HCV) NS3 protease is essential for viral replication. It has been a target of choice for intensive drug discovery research. On the basis of an active pentapeptide inhibitor, 1, we envisioned that macrocyclization from the P2 proline to P3 capping could enhance binding to the backbone Ala156 residue and the S4 pocket. Thus, a number of P2 proline-based macrocyclic {alpha}-ketoamide inhibitors were prepared and investigated in an HCV NS3 serine protease continuous assay (K*{sub i}). The biological activity varied substantially depending on factors such as the ring size, number of amino acid residues, number of methyl substituents, type of heteroatom in the linker, P3 residue, and configuration at the proline C-4 center. The pentapeptide inhibitors were very potent, with the C-terminal acids and amides being the most active ones (24, K*{sub i} = 8 nM). The tetrapeptides and tripeptides were less potent. Sixteen- and seventeen-membered macrocyclic compounds were equally potent, while fifteen-membered analogues were slightly less active. gem-Dimethyl substituents at the linker improved the potency of all inhibitors (the best compound was 45, K*{sub i} = 6 nM). The combination of tert-leucine at P3 and dimethyl substituents at the linker in compound 47 realized a selectivity of 307 against human neutrophil elastase. Compound 45 had an IC{sub 50} of 130 nM in a cellular replicon assay, while IC{sub 50} for 24 was 400 nM. Several compounds had excellent subcutaneous AUC and bioavailability in rats. Although tripeptide compound 40 was 97% orally bioavailable, larger pentapeptides generally had low oral bioavailability. The X-ray crystal structure of compounds 24 and 45 bound to the protease demonstrated the close interaction of the macrocycle with the Ala156 methyl group and S4 pocket. The strategy of macrocyclization has been proved to be successful in improving potency (>20-fold greater than that of 1) and in structural depeptization.

  10. Antiproton Accumulator (AA)

    Photographic Service


    The AA in its final stage of construction, before it disappeared from view under concrete shielding. Antiprotons were first injected, stochastically cooled and accumulated in July 1980. From 1981 on, the AA provided antiprotons for collisions with protons, first in the ISR, then in the SPS Collider. From 1983 on, it also sent antiprotons, via the PS, to the Low-Energy Antiproton Ring (LEAR). The AA was dismantled in 1997 and shipped to Japan.

  11. Produção de prolina e suscetibilidade ao glufosinato de amônio em plantas transgênicas de citrumelo Swingle Proline production by transgenic plants of Swingle citrumelo and susceptibility to glufosinate ammonium

    Cristine Elizabeth Alvarenga Carneiro


    Full Text Available O objetivo deste trabalho foi avaliar a sensibilidade de plantas transgênicas de citrumelo Swingle com elevada produção de prolina, ao herbicida glufosinato de amônio. As plantas utilizadas apresentavam a inserção do gene mutante da enzima delta1-pirrolina-5-carboxilato sintetase (P5CS, responsável pela biossíntese de prolina. A expressão do gene p5cs em plantas transgênicas causou aumento nas quantidades de prolina em tecidos foliares, em até cinco vezes, quando comparadas às plantas-controle tratadas com 200 µM de glufosinato de amônio. As plantas transgênicas acumularam maior quantidade de NH4+ nas folhas, em relação às plantas não-transgênicas. Os danos causados pelo herbicida foram avaliados in vitro, utilizando-se discos foliares cultivados em meio MS com diferentes concentrações de glufosinato de amônio. Observou-se maior clorose em discos foliares das plantas transgênicas, o que comprova a maior suscetibilidade de plantas de citrumelo Swingle com alta produção de prolina ao herbicida.The objective of this work was to evaluate the susceptibility to glufosinate ammonium of transgenic plants of Swingle citrumelo with high proline production. The mutant gene of the enzyme delta1-pyrroline-5-carboxylate synthetase (P5CS, the rate-limiting enzyme in proline biosynthesis, was inserted into Swingle citrumelo plants. The expression of the gene p5cs caused up to 5-fold increase on the proline content in leaf tissues of transgenic plants treated with 200 µM glufosinate ammonium, when compared with control plants. Leaves of transgenic plants accumulated higher amounts of NH4+ than the nontransgenic control. The herbicide toxicity was evaluated using leaf disks cultivated in MS medium, containing different concentrations of glufosinate ammonium. The severity of the chlorosis, observed in leaf disks of transgenic plants, confirmed the higher susceptibility of Swingle citrumelo plants, with high proline production, to this

  12. Whole genome phage display selects for proline-rich Boi polypeptides against Bem1p.

    Hertveldt, Kirsten; Robben, Johan; Volckaert, Guido


    Interaction selection by biopanning from a fragmented yeast proteome displayed on filamentous phage particles was successful in identifying proline-rich fragments of Boi1p and Boi2p. These proteins bind to the second "src homology region 3'' (SH3) domain of Bem1p, a protein of Saccharomyces cerevisiae involved in bud formation. Target Bem1p was a doubly-tagged recombinant, Bem1([Asn142-Ile551]), which strongly interacts in ELISA with a C-terminal 75 amino acids polypeptide from Cdc24p exposed on phage. The whole yeast genomic display library contained approximately 7.7 x 10(7) independent clones of sheared S. cerevisiae genomic DNA fused to a truncated M13 gene III. This study corroborates the value of fragmented-proteome display to identify strong and direct interacting protein modules.

  13. Conjugates of 1'-Aminoferrocene-1-carboxylic Acid and Proline: Synthesis, Conformational Analysis and Biological Evaluation

    Monika Kovačević


    Full Text Available Our previous studies showed that alteration of dipeptides Y-Fca-Ala-OMe (III into Y-Ala-Fca-OMe (IV (Y = Ac, Boc; Fca = 1'-aminoferrocene-1-carboxylic acid significantly influenced their conformational space. The novel bioconjugates Y-Fca-Pro-OMe (1, Y = Ac; 2, Y = Boc and Y-Pro-Fca-OMe (3, Y = Boc; 4, Y = Ac have been prepared in order to investigate the influence of proline, a well-known turn-inducer, on the conformational properties of small organometallic peptides with an exchanged constituent amino acid sequences. For this purpose, peptides 1–4 were subjected to detailed spectroscopic analysis (IR, NMR, CD spectroscopy in solution. The conformation of peptide 3 in the solid state was determined. Furthermore, the ability of the prepared conjugates to inhibit the growth of estrogen receptor-responsive MCF-7 mammary carcinoma cells and HeLa cervical carcinoma cells was tested.

  14. Epigallocatechin-3-gallate inhibits lactase but is alleviated by salivary proline-rich proteins.

    Naz, Shahina; Siddiqi, Rahmanullah; Dew, Tristan P; Williamson, Gary


    Lactase phlorizin hydrolase is a small intestinal brush border enzyme that catalyzes the hydrolysis of the milk sugar, lactose, and also many flavonoid glucosides. We demonstrate that epigallocatechin-3-gallate (EGCG), the principal flavonoid from green tea, inhibits in vitro hydrolysis of lactose by intestinal lactase. We then tested the hypothesis that salivary proline-rich proteins (PRPs) could modulate this inhibition and stabilize EGCG. Inhibition by EGCG of digestive enzymes (α-amylase>chymotrypsin>trypsin>lactase≫pepsin) was alleviated ∼2-6-fold by PRPs. Furthermore, PRPs appeared stable to proteolysis and also stabilized EGCG under digestive conditions in vitro. This is the first report on EGCG inhibition of lactase, and it quantifies the protective role of PRPs against EGCG inhibition of digestive enzymes.

  15. Synthesis, growth and characterization of a nonlinear optical crystal: Bis l-proline hydrogen nitrate.

    Selvaraju, K; Kirubavathi, K


    The single crystals of bis l-proline hydrogen nitrate (BLPHN) belonging to non-centrosymmetric space group were successfully grown by the slow evaporation solution growth technique. The BLPHN crystals of size 10×7×3mm(3) were obtained in 35days. Initially, the solubility tests were carried out for two solvents such as deionized water and mixed of deionized water-acetone. Among the two solvents, the solubility of BLPHN was found to be the highest in deionized water, so crystallization of BLPHN was done from its aqueous solution. As grown, crystals were characterized by single crystal X-ray diffraction studies and optical transmission spectral studies. Infrared spectroscopy, thermo gravimetric analysis and differential thermal analysis measurements were performed to study the molecular vibration and thermal behavior of the grown BLPHN crystals. Nonlinear optical (NLO) behavior of BLPHN crystal was studied by Kurtz and Perry powder method. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Ste20-related proline/alanine-rich kinase: A novel regulator of intestinal inflammation

    Yutao Yan; Didier Merlin


    Recently, inflammatory bowel disease (IBD) has been the subject of considerable research, with increasing attention being paid to the loss of intestinal epithelial cell barrier function as a mechanism of pathogenesis. Ste20-related proline/alanine-rich kinase (SPAK) is involved in regulating barrier function. SPAK is known to interact with inflammation-related kinases (such as p38, JNK, NKCC1, PKCθ, WNK and MLCK), and with transcription factor AP-1, resulting in diverse biological phenomena, including cell differentiation, cell transformation and proliferation, cytoskeleton rearrangement, and regulation of chloride transport. This review examines the involvement of Ste20-like kinases and downstream mitogen-activated protein kinases (MAPKs) pathways in the pathogenesis and control of intestinal inflammation. The primary focus will be on the molecular features of intestinal inflammation, with an emphasis on the interaction between SPAK and other molecules, and the effect of these interactions on homeostatic maintenance, cell volume regulation and increased cell permeability in intestinal inflammation.

  17. Evaluation of the effects of sensory denervation on osteoblasts by 3H-proline autoradiography.

    Chiego, D J; Singh, I J


    The inferior alveolar nerve was unilaterally resected in 30-day-old mice; other animals were unilaterally sham-operated. At 15, 30, 60, 90, or 150 days after surgery, the mice wee injected with 2 muCi of 3H-proline (sp. act. 1.0Ci/mM) per g of body weight and killed 15, 30, or 60 min later. Autoradiographs were prepared from 5 micron decalcified sagittal sections of mandibles and grain counts made over periosteal osteoblasts mesial to the first molar. In denervated mandibles, osteoblasts incorporated less isotope compared to controls with differences being maximal at the early intervals. These differences became attenuated with time, possibly due to an intrinsic compensatory mechanism, secondary to neurotrophic regulation.

  18. Susceptibility to Dental Caries and the Salivary Proline-Rich Proteins

    Martin Levine


    Full Text Available Early childhood caries affects 28% of children aged 2–6 in the US and is not decreasing. There is a well-recognized need to identify susceptible children at birth. Caries-free adults neutralize bacterial acids in dental biofilms better than adults with severe caries. Saliva contains acidic and basic proline-rich proteins (PRPs which attach to oral streptococci. The PRPs are encoded within a small region of chromosome 12. An acidic PRP allele (Db protects Caucasian children from caries but is more common in African Americans. Some basic PRP allelic phenotypes have a three-fold greater frequency in caries-free adults than in those with severe caries. Early childhood caries may associate with an absence of certain basic PRP alleles which bind oral streptococci, neutralize biofilm acids, and are in linkage disequilibrium with Db in Caucasians. The encoding of basic PRP alleles is updated and a new technology for genotyping them is described.

  19. Determination of proline in honey: comparison between official methods, optimization and validation of the analytical methodology.

    Truzzi, Cristina; Annibaldi, Anna; Illuminati, Silvia; Finale, Carolina; Scarponi, Giuseppe


    The study compares official spectrophotometric methods for the determination of proline content in honey - those of the International Honey Commission (IHC) and the Association of Official Analytical Chemists (AOAC) - with the original Ough method. Results show that the extra time-consuming treatment stages added by the IHC method with respect to the Ough method are pointless. We demonstrate that the AOACs method proves to be the best in terms of accuracy and time saving. The optimized waiting time for the absorbance recording is set at 35min from the removal of reaction tubes from the boiling bath used in the sample treatment. The optimized method was validated in the matrix: linearity up to 1800mgL(-1), limit of detection 20mgL(-1), limit of quantification 61mgL(-1). The method was applied to 43 unifloral honey samples from the Marche region, Italy. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. [Determination of underivatized glycine and proline in vinegar turtle shell by HPLC-ELSD].

    Wang, Xinyu; Tan, Xiaomei; Gao, Mingze; Peng, Jinlong


    To establish a method to determine the underivatized glycine (Gly) and proline (Pro) in vinegar turtle shell. An HPLC-ELSD method was conducted on a Prevail C18 column (4.6 mm x 250 mm, 5 microm) with the mobile phase of acetonitrile and 0.7% trifluoroacetic acid solution (containing 5.0 mmol x L(-1) heptafluorobutyric acid), and elution time was 15 min. The calibration curves were showed good linearity within the concentration range of 0.14-0.6 g x L(-1). The average recoveries were 101.2% and 102.5%, and RSD were 1.9% and 2.5%, respectively. Since this method needs neither the special amino acid analyzer nor derivation of amino acid., it is efficient, simple and accurate., which could be used for quality control of vinegar turtle shell.

  1. Preparation of Polyaminopyridines Using a CuI/l-Proline-Catalyzed C-N Polycoupling Reaction

    Paulo C. M. L. Miranda


    Full Text Available Polyaminopyridines (PAPy were chemically prepared from amino-bromopyridines by a CuI/l-proline-catalyzed C-N polycondensation reaction. The formation of the polymer was confirmed by GPC, XRD, XRF, FTIR, UV-vis (λmax = 400 nm, 1H and 13C NMR. The number-average molecular weights (Mn were estimated by end-group analysis using X-ray fluorescence (up to 6000 Da. TGA analysis of PAPy with higher Mn showed greater thermal stability up to 170 oC. Viscosity measurements of polymer in formic acid at 30 oC indicated a polyelectrolyte nature of PAPy solutions. Furthermore, the amorphicity of the material was observed by X-ray diffraction analysis.

  2. A proline-based aminophenol ligand: synthesis, iron complexation, magnetic, electronic and redox investigation.

    Sheykhi, Hamid; Safaei, Elham


    A new proline-based aminophenol ligand was synthesized by a convenient procedure. The ligand was characterized by (1)H NMR, (13)C NMR and IR spectroscopies, elemental analysis and optical activity measurements. Mononuclear iron(III) complex (FeL(Pro)) of this ligand was synthesized and characterized by IR, UV-vis, ESI-MS, magnetic susceptibility studies and cyclic voltammetry techniques. The equilibrium formation constant of FeL(Pro) and the pure UV-vis spectral profile of the complex was determined by multivariate hard modeling method. The molecular structure of FeL(Pro) determined by ESI-MS consist of two aminophenolate ligands. The variation of magnetic susceptibility with temperature indicates paramagnetic iron(III) in the monomeric complex. FeL(Pro) complex undergo metal-centered reduction, and ligand-centered oxidation. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. A Novel Vasoactive Proline-Rich Oligopeptide from the Skin Secretion of the Frog Brachycephalus ephippium.

    Daniel Dias Rufino Arcanjo

    Full Text Available Proline-rich oligopeptides (PROs are a large family which comprises the bradykinin-potentiating peptides (BPPs. They inhibit the activity of the angiotensin I-converting enzyme (ACE and have a typical pyroglutamyl (Pyr/proline-rich structure at the N- and C-terminus, respectively. Furthermore, PROs decrease blood pressure in animals. In the present study, the isolation and biological characterization of a novel vasoactive BPP isolated from the skin secretion of the frog Brachycephalus ephippium is described. This new PRO, termed BPP-Brachy, has the primary structure WPPPKVSP and the amidated form termed BPP-BrachyNH2 inhibits efficiently ACE in rat serum. In silico molecular modeling and docking studies suggest that BPP-BrachyNH2 is capable of forming a hydrogen bond network as well as multiple van der Waals interactions with the rat ACE, which blocks the access of the substrate to the C-domain active site. Moreover, in rat thoracic aorta BPP-BrachyNH2 induces potent endothelium-dependent vasodilatation with similar magnitude as captopril. In DAF-FM DA-loaded aortic cross sections examined by confocal microscopy, BPP-BrachyNH2 was found to increase the release of nitric oxide (NO. Moreover, BPP-BrachyNH2 was devoid of toxicity in endothelial and smooth muscle cell cultures. In conclusion, the peptide BPP-BrachyNH2 has a novel sequence being the first BPP isolated from the skin secretion of the Brachycephalidae family. This opens for exploring amphibians as a source of new biomolecules. The BPP-BrachyNH2 is devoid of cytotoxicity and elicits endothelium-dependent vasodilatation mediated by NO. These findings open for the possibility of potential application of these peptides in the treatment of endothelial dysfunction and cardiovascular diseases.

  4. Control of embryonic stem cell metastability by L-proline catabolism

    Laura Casalino; Stefania Comes; Giuseppina Lambazzi; Benedetta De Stefano; Stefania Filosa; Sandro De Falco; Dario De Cesare; Gabriella Minchiotti; Eduardo Jorge Patriarca


    The molecular mechanisms controlling mouse embryonic stem cell (ESC) metastability, i.e. their capacity to fluctuate between different states of pluripotency, are not fully resolved. We developed and used a novel automation platform, the Cellmaker, to screen a library of metabolites on two ESC-based phenotypic assays (i.e. proliferation and colony phenotype) and identified two metabolically related amino acids, namely L-proline (L-Pro) and L-ornithine (L-Orn), as key regulators of ESC metastability. Both compounds,but mainly L-Pro, force ESCs toward a novel epiblast stem cell (EpiSC)-like state, in a dose- and time-dependent manner. Unlike EpiSCs, L-Pro-induced cells (PiCs) contribute to chimeric embryos and rely on leukemia inhibitor factor (LIF) to self-renew.Furthermore, PiCs revert to ESCs or differentiate randomly upon removal of either L-Pro or LIF, respectively. Remarkably, PiC generation depends on both L-Pro metabolism (uptake and oxidation) and Fgf5 induction, and is strongly counteracted by antioxidants,mainly L-ascorbic acid (vitamin C, Vc). ESCs (←→) PiCs phenotypic transition thus represents a previously undefined dynamic equilibrium between pluripotent states, which can be unbalanced either toward an EpiSC-like or an ESC phenotype by L-Pro/L-Orn or Vc treatments, respectively. All together, our data provide evidence that E5C metastability can be regulated at a metabolic level.Kevwords: embryonic stem cells, L-proline, vitamin C, colony phenotype, pluripotent states, metastability

  5. Versatile three-component procedure for combinatorial synthesis of spiro-oxindoles with fused chromenes catalysed by L-proline

    Majid M Heravi; Masoumeh Zakeri; Arezoo Moharami


    An efficient synthesis of spiro-oxindoles is accomplished by a one-pot three-component condensation of isatin, malononitrile or cyanoacetic ester and naphthol in the presence of L-proline as a catalyst. This method is of great value because of its shorter reaction times, high yield, and easy processing.

  6. Effect of Salinity Stress on Morphological and Proline Content of Eight Landraces Fenugreek (Trigonella foenum - graecum L.

    H Farhadi


    Full Text Available In order to evaluate the effect of salinity on some morphological characteristics and proline content of eight fenugreek landraces and identification of the best landrace, a factorial experiment was conducted on the basis of complete randomized design with three replicates in the research field of Ferdowsi University of Mashhad in 2013. Experimental treatments were combination of eight fenugreek landrace (Isfahan, Tabriz, Hamedan, Sari, Challous, Amol, Mashhad and Yasooj and four levels of salinity stress (0, 60, 120 and 180 Mm NaCl. The ANOVA results revealed the significant effect of salinity on plant height, number of branches/plant, number of nodes, inter nodal distance, root length, shoot length, root dry weight, shoot dry weight, fresh weight of fruit, nut and proline content. The highest level of salinity (180 mM NaCl significantly decreased the mentioned plant characters by 16.72%, 30.44%, 18.22%, 49.45%, 11.95%, 13%, 48.44%, 57.90%, 59.56%, 54.11% compared to control respectively. Proline content in the highest salinity level (180 mM NaCl was increased by 44.57% compared to control. The greatest amount of shoot vegetative yield was obtained from control (without salinity and the highest rate of proline was achieved from 180 Mm treatment.

  7. L-proline-catalysed synthesis of functionalized unsymmetrical dihydro-1H-indeno[1,2-b]pyridines

    Farahnaz Kargar Behbahani; Hourieh Sadat Alaei


    Aromatic aldehydes have been employed in a one-pot four-component reaction with ethyl acetoacetate, 1,3-indandione and ammonium acetate in the presence of L-proline in water and under reflux condition to afford the corresponding dihydro-1H-indeno[1,2-b]pyridines in very good yields.

  8. Kinetics and Mechanism for Oxidation of L-Proline by Bis(hydrogen periodato) argentate (Ⅲ) Complex Anion

    SUN Han-Wen; SHI Hong-Mei; SHEN Shi-Gang; FANG Wei-Jun; GUO Zhi-Feng


    Oxidation of L-proline by bis(hydrogen periodato)argentate(Ⅲ) complex anion, [Ag(HIO6)2]5-, has been studied in aqueous alkaline medium in the temperature range of 25-40 ℃ by use of conventional spectrophotometry.Under the conditions for kinetic measurements, the oxidation results in decarboxylation of proline, giving rise to γ-aminobutyrate as identified by mass spectrometry. The oxidation kinetics is first order with respect to the silver(Ⅲ)and proline concentrations; the second-order rate constants, decreasing with increasing [periodate], are essentially independent of [OH-]. The kinetic results were interpreted in terms of a reaction mechanism which involves a pre-equilibrium between [Ag(HIO6)2]5- and [Ag(HIO6)(H2O)(OH)]2-, a mono-periodate coordinated silver(Ⅲ)complex. Both Ag(Ⅲ) complexes are reduced parallelly by the fully deprotonated form of proline in reductant and shows a lower reactivity than [Ag(HIO6)(OH)(H2O)]2-.

  9. Unique proline-benzoquinone pigment from the colored nectar of "bird's Coca cola tree" functions in bird attractions.

    Luo, Shi-Hong; Liu, Yan; Hua, Juan; Niu, Xue-Mei; Jing, Shu-Xi; Zhao, Xu; Schneider, Bernd; Gershenzon, Jonathan; Li, Sheng-Hong


    The major pigment responsible for the dark brown nectar of the "bird's Coca cola tree", Leucosceptrum canum (Labiatae), was isolated and identified as a unique symmetric proline-quinone conjugate, 2,5-di-(N-(-)-prolyl)-para-benzoquinone (DPBQ). Behavioral experiments with both isolated and synthetic authentic samples indicated that DPBQ functions mainly as a color attractant to bird pollinators.

  10. Antioxidant properties of small proline-rich proteins : from epidermal cornification to global ROS detoxification and wound healing

    Vermeij, Wilbert Peter


    The small proline-rich (SPRR) proteins are generally known for their involvement in the formation and adaptation of the skin’s barrier. During the cornification process, they are cross-linked within the cornified cell envelope (CE) and as such they are responsible for the physical and permeability

  11. Structure of the WW domain of a kinase-associated protein complexed with a proline-rich peptide.

    Macias, M J; Hyvönen, M; Baraldi, E; Schultz, J; Sudol, M; Saraste, M; Oschkinat, H


    The WW domain is a new protein module with two highly conserved tryptophans that binds proline-rich peptide motifs in vitro. It is present in a number of signalling and regulatory proteins, often in several copies. Here we investigate the solution structure of the WW domain of human YAP65 (for Yes kinase-associated protein) in complex with proline-rich peptides containing the core motif PPxY. The structure of the domain with the bound peptide GTPPPPYTVG is a slightly curved, three-stranded, antiparallel beta-sheet. Two prolines pack against the first tryptophan, forming a hydrophobic buckle on the convex side of the sheet. The concave side has three exposed hydrophobic residues (tyrosine, tryptophan and leucine) which form the binding site for the ligand. A non-conserved isoleucine in the amino-terminal flanking region covers a hydrophobic patch and stabilizes the WW domain of human YAP65 in vitro. The structure of the WW domain differs from that of the SH3 domain and reveals a new design for a protein module that uses stacked aromatic surface residues to arrange a binding site for proline-rich peptides.

  12. Effects of metabolic neuropeptides from insect corpora cardiaca on proline metabolism of the African fruit beetle, Pachnoda sinuata.

    Auerswald, L; Gäde, G


    The effect of neuropeptides from the corpora cardiaca of the fruit beetle Pachnoda sinuata on proline metabolism has been investigated in vivo. Conspecific injections of a crude extract from corpora cardiaca cause an increase of the concentration of proline in the haemolymph by nearly 20% and a decrease of the concentration of alanine, the precursor in proline synthesis, by about 64% when compared with a water-injected group. Purification of an extract of corpora cardiaca on reversed-phase liquid chromatography revealed two distinct UV absorbance and fluorescence peaks that cause hyperprolinaemia in the fruit beetle. The major peak is the previously identified octapeptide Mem-CC; the second peak is also a peptide, but its primary sequence remains, as yet, unidentified. Synthetic Mem-CC elicited time- and dose-dependent increases/decreases of the concentrations of proline and alanine in the haemolymph respectively. Furthermore, the receptor for this peptide seems to be specific in P. sinuata: only peptides of the large family of adipokinetic hormones with an Asp, Asn or Gly residue at position 7 could elicit biological activity, whereas those with a Trp, Ser or Val residue at this position did not have any activity.

  13. Highly enantioselective synthesis of fluorinated gamma-amino alcohols through proline-catalyzed cross-Mannich reaction.

    Fustero, Santos; Jiménez, Diego; Sanz-Cervera, Juan F; Sánchez-Roselló, María; Esteban, Elisabet; Simón-Fuentes, Antonio


    A new, simple route for the synthesis of fluorinated beta-alkyl gamma-amino alcohols in optically pure form in only two steps and featuring proline catalysis from inexpensive and readily available starting materials is described. The applied strategy allows for the introduction of diversity into both the beta-fluoroalkyl and alpha-alkyl groups of these compounds. [reaction: see text

  14. Antioxidant properties of small proline-rich proteins : from epidermal cornification to global ROS detoxification and wound healing

    Vermeij, Wilbert Peter


    The small proline-rich (SPRR) proteins are generally known for their involvement in the formation and adaptation of the skin’s barrier. During the cornification process, they are cross-linked within the cornified cell envelope (CE) and as such they are responsible for the physical and permeability b

  15. Role of the Escherichia coli SbmA in the antimicrobial activity of proline-rich peptides.

    Mattiuzzo, Maura; Bandiera, Antonella; Gennaro, Renato; Benincasa, Monica; Pacor, Sabrina; Antcheva, Nikolinka; Scocchi, Marco


    In contrast to many antimicrobial peptides, members of the proline-rich group of antimicrobial peptides inactivate Gram-negative bacteria by a non-lytic mechanism. Several lines of evidence indicate that they are internalized into bacteria and their activity mediated by interaction with unknown cellular components. With the aim of identifying such interactors, we selected mutagenized Escherichia coli clones resistant to the proline-rich Bac7(1-35) peptide and analysed genes responsible for conferring resistance, whose products may thus be involved in the peptide's mode of action. We isolated a number of genomic regions bearing such genes, and one in particular coding for SbmA, an inner membrane protein predicted to be part of an ABC transporter. An E. coli strain carrying a point mutation in sbmA, as well as other sbmA-null mutants, in fact showed resistance to several proline-rich peptides but not to representative membranolytic peptides. Use of fluorescently labelled Bac7(1-35) confirmed that resistance correlated with a decreased ability to internalize the peptide, suggesting that a bacterial protein, SbmA, is necessary for the transport of, and for susceptibility to, proline-rich antimicrobial peptides of eukaryotic origin.

  16. Accumulator-Based Deep-Sea Microbe Gastight Sampling Technique

    HUANG Zhong-hua; LIU Shao-jun; JIN Bo


    The accumulator is used as a pressure compensation device to realize deep-sea microbe gastight sampling. Four key states of the accumulator are proposed to describe the pressure compensation process and a corresponding mathematical model is established to investigate the relationship between the results of pressure compensation and the parameters of the accumulator. Simulation results show that during the falling process of the sampler, the accumulator's real opening pressure is greater than its precharge pressure; when the sampling depth is 6000 m and the accumulator's precharge pressure is less than 30 MPa, to increase the accumulator's precharge pressure can improve pressure compensation results obviously. Laboratory experiments at 60 MPa show that the accumulator is an effective and reliable pressure compensation device for deep-sea microbe samplers. The success in sea trial at a depth of 2000 m in the South China Sea shows that the mathematical model and laboratory experiment results are reliable.

  17. Naturally evolved enhanced Cd tolerance of Dianthus carthusianorum L. is not related to accumulation of thiol peptides and organic acids.

    Wójcik, Małgorzata; Dresler, Sławomir; Plak, Andrzej; Tukiendorf, Anna


    Two contrasting ecotypes of Dianthus carthusianorum L., metallicolous (M) and nonmetallicolous (NM), were cultivated in hydroponics at 0-50 μM Cd for 14 days to compare their Cd accumulation, sensitivity and tolerance mechanisms. While both ecotypes contained similar concentrations of Cd in the shoots and roots, the M ecotype was more Cd-tolerant (as measured by fresh weight production and root and leaf viability). Both ecotypes accumulated phytochelatins (PCs) in response to Cd with a higher amount thereof found in the NM ecotype. Concentrations of PCs remained unchanged with increasing Cd concentrations in the root tissues, but their content in the shoots increased. The addition of L-buthionine-sulfoximine (BSO) diminished glutathione (GSH) accumulation and arrested PC production, which increased the sensitivity to Cd of the NM, but not M ecotype. Organic acids (malate and citrate) as well as proline accumulation did not change significantly after Cd exposition and was at the same level in both ecotypes. The enhanced Cd tolerance of the M ecotype of D. carthusianorum cannot be explained in terms of restricted Cd uptake and differential production of PCs, organic acids or proline; some other mechanisms must be involved in its adaptation to the high Cd content in the environment.

  18. Actions of a proline analogue, L-thiazolidine-4-carboxylic acid (T4C, on Trypanosoma cruzi.

    Anahí Magdaleno

    Full Text Available It is well established that L-proline has several roles in the biology of trypanosomatids. In Trypanosoma cruzi, the etiological agent of Chagas' disease, this amino acid is involved in energy metabolism, differentiation processes and resistance to osmotic stress. In this study, we analyzed the effects of interfering with L-proline metabolism on the viability and on other aspects of the T. cruzi life cycle using the proline analogue L- thiazolidine-4-carboxylic acid (T4C. The growth of epimastigotes was evaluated using different concentrations of T4C in standard culture conditions and at high temperature or acidic pH. We also evaluated possible interactions of this analogue with stress conditions such as those produced by nutrient starvation and oxidative stress. T4C showed a dose-response effect on epimastigote growth (IC(50 = 0.89+/-0.02 mM at 28 degrees C, and the inhibitory effect of this analogue was synergistic (p<0.05 with temperature (0.54+/-0.01 mM at 37 degrees C. T4C significantly diminished parasite survival (p<0.05 in combination with nutrient starvation and oxidative stress conditions. Pre-incubation of the parasites with L-proline resulted in a protective effect against oxidative stress, but this was not seen in the presence of the drug. Finally, the trypomastigote bursting from infected mammalian cells was evaluated and found to be inhibited by up to 56% when cells were treated with non-toxic concentrations of T4C (between 1 and 10 mM. All these data together suggest that T4C could be an interesting therapeutic drug if combined with others that affect, for example, oxidative stress. The data also support the participation of proline metabolism in the resistance to oxidative stress.

  19. Biomass Yield and Steviol Glycoside Production in Callus and Suspension Culture of Stevia rebaudiana Treated with Proline and Polyethylene Glycol.

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay


    Enhanced production of steviol glycosides (SGs) was observed in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol (PEG). To study their effect, yellow-green and compact calli obtained from in vitro raised Stevia leaves were sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of proline (2.5-10 mM) and PEG (2.5-10 %) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension culture biomass (i.e. both fresh and dry weight content) was increased with 5 mM proline and 5 % PEG, while at further higher concentrations, they got reduced. Further, quantification of SGs content in callus (collected at 15th day) and suspension culture (collected at 10th and 15th day) treated with and without elicitors was analysed by HPLC. It was observed that chemical stress enhanced the production of SGs significantly. In callus, the content of SGs increased from 0.27 (control) to 1.09 and 1.83 % with 7.5 mM proline and 5 % PEG, respectively, which was about 4.0 and 7.0 times higher than control. However, in the case of suspension culture, the same concentrations of proline and polyethylene glycol enhanced the SG content from 1.36 (control) to 5.03 and 6.38 %, respectively, on 10th day which were 3.7 times and 4.7 times higher than control.

  20. Synthesis of fluorescent label, DBD-beta-proline, and the resolution efficiency for chiral amines by reversed-phase chromatography.

    Min, Jun Zhe; Toyo'oka, Toshimasa; Kato, Masaru; Fukushima, Takeshi


    DBD-d(and l)-beta-proline, new fluorescent chiral derivatization reagents, were synthesized from the reaction of 4-(N,N-dimethylaminosulfonyl)-7- fl uoro-2,1,3-benzoxadiazole (DBD-F) with beta-proline. The racemic mixture synthesized was separated by a chiral stationary phase (CSP) column, Chiralpak AD-H, with n-hexane-EtOH-TFA-diethylamine (70:30:0.1:0.1) as the mobile phase. The dl-forms were decided according to the results obtained from a circular dichroism (CD) detector after separation by the CSP column. The fractionated enantiomers reacted with chiral amine to produce a couple of diastereomers. The labeling proceeded in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and pyridine as the activation reagents. The reaction conditions were mild and no racemization occurred during the diastereomer formation. The resulting diastereomers fluoresced at around 570 nm (excitation at around 460 nm). Good linearity of the calibration curves was obtained in the range 1-75 pmol and the detection limits on chromatogram were less than 1 pmol. The separability of the diastereomers was compared with the diastereomers derived from DBD-d(or l)-proline. The resolution values (Rs) obtained from the diastereomers of three chiral amines with DBD-d(or l)-beta-proline were higher than those derived from DBD-d(or l)-proline, e.g. dl-phenylalanine methylester (dl-PAME), 2.23 vs 1.37; (R)(S)-1-phenylethylamine [(R)(S)-PEA], 2.09 vs 1.13; and (R)(S)-1-(1-naphthyl)ethylamines [(R)(S)-NEA], 5.19 vs 1.23. The results suggest that the position of COOH group on pyrrolidine moiety in the structures is one of the important factors for the efficient separation of a couple of the diastereomers.

  1. New proline-rich oligopeptides from the venom of African adders: Insights into the hypotensive effect of the venoms.

    Kodama, Roberto T; Cajado-Carvalho, Daniela; Kuniyoshi, Alexandre K; Kitano, Eduardo S; Tashima, Alexandre K; Barna, Barbara F; Takakura, Ana Carolina; Serrano, Solange M T; Dias-Da-Silva, Wilmar; Tambourgi, Denise V; Portaro, Fernanda V


    The snakes from the Bitis genus are some of the most medically important venomous snakes in sub Saharan Africa, however little is known about the composition and effects of these snake venom peptides. Considering that the victims with Bitis genus snakes have exacerbate hypotension and cardiovascular disorders, we investigated here the presence of angiotensin-converting enzyme modulators on four different species of venoms. The peptide fractions from Bitis gabonica gabonica, Bitis nasicornis, Bitis gabonica rhinoceros and Bitis arietans which showed inhibitory activity on angiotensin-converting enzyme were subjected to mass spectrometry analysis. Eight proline-rich peptides were synthetized and their potencies were evaluated in vitro and in vivo. The MS analysis resulted in over 150 sequences, out of which 32 are new proline-rich oligopeptides, and eight were selected for syntheses. For some peptides, inhibition assays showed inhibitory potentials of cleavage of angiotensin I ten times greater when compared to bradykinin. In vivo tests showed that all peptides decreased mean arterial pressure, followed by tachycardia in 6 out of 8 of the tests. We describe here some new and already known proline-rich peptides, also known as bradykinin-potentiating peptides. Four synthetic peptides indicated a preferential inhibition of angiotensin-converting enzyme C-domain. In vivo studies show that the proline-rich oligopeptides are hypotensive molecules. Although proline-rich oligopeptides are known molecules, we present here 32 new sequences that are inhibitors of the angiotensin-converting enzyme and consistent with the symptoms of the victims of Bitis spp, who display severe hypotension. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Identification and characterization of bifunctional proline racemase/hydroxyproline epimerase from archaea: discrimination of substrates and molecular evolution.

    Seiya Watanabe

    Full Text Available Proline racemase (ProR is a member of the pyridoxal 5'-phosphate-independent racemase family, and is involved in the Stickland reaction (fermentation in certain clostridia as well as the mechanisms underlying the escape of parasites from host immunity in eukaryotic Trypanosoma. Hydroxyproline epimerase (HypE, which is in the same protein family as ProR, catalyzes the first step of the trans-4-hydroxy-L-proline metabolism of bacteria. Their substrate specificities were previously considered to be very strict, in spite of similarities in their structures and catalytic mechanisms, and no racemase/epimerase from the ProR superfamily has been found in archaea. We here characterized the ProR-like protein (OCC_00372 from the hyperthermophilic archaeon, Thermococcus litoralis (TlProR. This protein could reversibly catalyze not only the racemization of proline, but also the epimerization of 4-hydroxyproline and 3-hydroxyproline with similar kinetic constants. Among the four (putative ligand binding sites, one amino acid substitution was detected between TlProR (tryptophan at the position of 241 and natural ProR (phenylalanine. The W241F mutant showed a significant preference for proline over hydroxyproline, suggesting that this (hydrophobic and bulky tryptophan residue played an importance role in the recognition of hydroxyproline (more hydrophilic and bulky than proline, and substrate specificity for hydroxyproline was evolutionarily acquired separately between natural HypE and ProR. A phylogenetic analysis indicated that such unique broad substrate specificity was derived from an ancestral enzyme of this superfamily.

  3. Nitrate deficiency reduces cadmium and nickel accumulation in chamomile plants.

    Kovácik, Jozef; Klejdus, Borivoj; Stork, Frantisek; Hedbavny, Josef


    The effect of nitrogen (nitrate) deficiency (-N) on the accumulation of cadmium (Cd) and nickel (Ni) in chamomile ( Matricaria chamomilla ) plants was studied. Elimination of N from the culture medium led to decreases in N-based compounds (free amino acids and soluble proteins) and increases in C-based compounds (reducing sugars, soluble phenols, coumarins, phenolic acids, and partially flavonoids and lignin), being considerably affected by the metal presence. Proline, a known stress-protective amino acid, decreased in all -N variants. The activity of phenylalanine ammonia-lyase was stimulated only in -N control plants, whereas the activities of polyphenol oxidase and guaiacol peroxidase were never reduced in -N variants in comparison with respective +N counterparts. Among detected phenolic acids, chlorogenic acid strongly accumulated in all N-deficient variants in the free fraction and caffeic acid in the cell wall-bound fraction. Mineral nutrients were rather affected by a given metal than by N deficiency. Shoot and total root Cd and Ni amounts decreased in -N variants. On the contrary, ammonium-fed plants exposed to N deficiency did not show similar changes in Cd and Ni contents. The present findings are discussed with respect to the role of phenols and mineral nutrition in metal uptake.

  4. Irrigation with industrial wastewater activates antioxidant system and osmoprotectant accumulation in lettuce, turnip and tomato plants.

    Hashem, H A; Hassanein, R A; El-Deep, M H; Shouman, A I


    We focused on the impact of industrial wastes on the water quality of the El-Amia drain in Egypt and the effect of irrigation with industrial wastewater on the growth, cell membranes, photosynthetic pigment content, the antioxidant system and selected osmoprotectants (proline, total amino nitrogen and soluble sugars) in three crop plants: turnip, tomato and lettuce. Furthermore, the present work focused on the analysis of the heavy metal content and its accumulation in the studied plants. For this purpose, water samples were collected 1, 10 and 19 km from the beginning of the drain and used for irrigation, with fresh water as a control. We found that industrial wastewater contained significant amounts of heavy metals (Cd, Ni and Co) warranted a pollution problem as their amounts exceed the maximum recommended concentrations according to FAO guidelines for trace metals in irrigation water. The three crop plants accumulate significant amounts of heavy metals in their shoots and roots and showed a significant decrease in leaf area, fresh weight and dry weight of shoots and roots, accompanied by a marked reduction in photosynthetic pigment content and damage to cell membranes, as indicated by increased electrolyte leakage and a lower membrane stability index. Significant increases in the activities of antioxidant enzymes and in the glutathione, proline, soluble sugar and total amino nitrogen content in response to irrigation with wastewater may be defense mechanisms induced in response to heavy metal stress.

  5. Ice slurry accumulation

    Christensen, K.G.; Kauffeld, M.


    More and more refrigeration systems are designed with secondary loops, thus reducing the refrigerant charge of the primary refrigeration plant. In order not to increase energy consumption by introducing a secondary refrigerant, alternatives to the well established single phase coolants (brines) and different concepts of the cooling plant have to be evaluated. Combining the use of ice-slurry - mixture of water, a freezing point depressing agent (antifreeze) and ice particles - as melting secondary refrigerant and the use of a cool storage makes it possible to build plants with secondary loops without increasing the energy consumption and investment. At the same time the operating costs can be kept at a lower level. The accumulation of ice-slurry is compared with other and more traditional storage systems. The method is evaluated and the potential in different applications is estimated. Aspects of practically use of ice-slurry has been examined in the laboratory at the Danish Technological Institute (DTI). This paper will include the final conclusions from this work concerning tank construction, agitator system, inlet, outlet and control. The work at DTI indicates that in some applications systems with ice-slurry and accumulation tanks have a great future. These applications are described by a varying load profile and a process temperature suiting the temperature of ice-slurry (-3 - -8/deg. C). (au)

  6. Determination of the cis-trans isomerization barriers of L-alanyl-L-proline in aqueous solutions and at water/hydrophobic interfaces by on-line temperature-jump relaxation HPLC and dynamic on-column reaction HPLC.

    Shibukawa, Masami; Miyake, Ayaka; Eda, Sayaka; Saito, Shingo


    Proline cis-trans isomerization is known to play a key role in the rate-determining steps of protein folding. It is thus very important to understand the influence of environments, not only bulk solutions but also microenvironments such as interfaces, on the isomerization reaction of proline peptides. Here we present two HPLC methods for measurements of kinetic and equilibrium parameters for the isomerization reactions in bulk solutions and at liquid/solid interfaces. On-line temperature-jump relaxation HPLC (T-jump HPLC) allows the determination of forward and reverse rate constants of the isomerization in a bulk solution by monitoring the whole time course of conversion of pure isomers from both sides of the reaction, in contrast to other HPLC and capillary zone electrophoresis as well as spectrometric and calorimetric methods, which use a mixture of the isomers. We can then determine cis-trans isomerization barriers of the peptide at liquid/solid interfaces from the kinetic data obtained by dynamic on-column reaction HPLC and T-jump HPLC. We observed that the interconversion around the peptide bond for l-alanyl-l-proline (Ala-Pro) in water is accelerated at the surfaces of an alkyl-bonded silica and a poly(styrene-divinylbenzene) copolymer resin, and this is caused by a remarkable decrease in the enthalpy of activation. The molecular structures of the cis and trans forms of Ala-Pro estimated by quantum mechanics calculation reveal that an equilibrium shift toward the cis form as well as the rapid isomerization of Ala-Pro at the water/hydrophobic interfaces can be attributed to the lower polarity of the interfacial water at the surfaces of the hydrophobic materials compared to that of bulk water.

  7. Proline induces the expression of salt-stress-responsive proteins and may improve the adaptation of Pancratium maritimum L. to salt-stress.

    Khedr, Abdel Hamid A; Abbas, Mohammad A; Wahid, Amal A Abdel; Quick, W Paul; Abogadallah, Gaber M


    Proline is an important component of salt-stress responses of plants. In this study the role of proline as part of salt-stress signalling in the desert plant Pancratium maritimum L. was examined. The data showed that salt-stress brought about a reduction of the growth and protein content, particularly at 300 mM NaCl, that was significantly increased by exogenous proline. In the leaves, salt-stress up-regulated ubiquitin, a small protein targeting damaged proteins for degradation via the proteasome, up to 5-fold as detected by western blotting. This change was also affected by proline even in non-stressed leaves. However, salt-stress resulted in a decrease in the amount of ubiquitin-conjugates, particularly in the roots, and this effect was reversed by exogenous proline. Severe salt-stress resulted in an inhibition of the antioxidative enzymes catalase and peroxidase as revealed by spectrophotometric assays and activity gels, but the activity of these enzymes was also maintained significantly higher in the presence of proline. Salt-stress also up-regulated several dehydrin proteins, analysed by western blotting, even in non-stressed plants. It is concluded that proline improves the salt-tolerance of Pancratium maritimum L. by protecting the protein turnover machinery against stress-damage and up-regulating stress protective proteins.

  8. Effect of Biochar on Relieving Cadmium Stress and Reducing Accumulation in Super japonica Rice

    ZHANG Zhen-yu; MENG Jun; DANG Shu; CHEN Wen-fu


    It is of great importance to solve the threats induced by cadmium pollution on crops. This paper examined the effect of biochar on cadmium accumulation in japonica rice and revealed the mechanism underlying the response of protective enzyme system to cadmium stress. Biochar derived from rice straw was applied at two application rates under three cadmium concentrations. Shennong 265, super japonica rice variety, was selected as the test crop. The results indicated that cadmium content in above-ground biomass of rice increased with increasing soil cadmium concentrations, but the biochar application could suppress the accumulation of cadmium to some extent. Under high concentrations of cadmium, content of free proline and MDA (malondialdehyde) were high, so did the SOD (superoxide dismutase), POD (peroxidase) and CAT (catalase) activity in the lfag leaf of rice. However, the protective enzyme activities remained at low level when biochar was added.

  9. Solids Accumulation Scouting Studies

    Duignan, M. R.; Steeper, T. J.; Steimke, J. L.


    The objective of Solids Accumulation activities was to perform scaled testing to understand the behavior of remaining solids in a Double Shell Tank (DST), specifically AW-105, at Hanford during multiple fill, mix, and transfer operations. It is important to know if fissionable materials can concentrate when waste is transferred from staging tanks prior to feeding waste treatment plants. Specifically, there is a concern that large, dense particles containing plutonium could accumulate in poorly mixed regions of a blend tank heel for tanks that employ mixing jet pumps. At the request of the DOE Hanford Tank Operations Contractor, Washington River Protection Solutions, the Engineering Development Laboratory of the Savannah River National Laboratory performed a scouting study in a 1/22-scale model of a waste staging tank to investigate this concern and to develop measurement techniques that could be applied in a more extensive study at a larger scale. Simulated waste tank solids: Gibbsite, Zirconia, Sand, and Stainless Steel, with stainless steel particles representing the heavier particles, e.g., plutonium, and supernatant were charged to the test tank and rotating liquid jets were used to mix most of the solids while the simulant was pumped out. Subsequently, the volume and shape of the mounds of residual solids and the spatial concentration profiles for the surrogate for heavier particles were measured. Several techniques were developed and equipment designed to accomplish the measurements needed and they included: 1. Magnetic particle separator to remove simulant stainless steel solids. A device was designed and built to capture these solids, which represent the heavier solids during a waste transfer from a staging tank. 2. Photographic equipment to determine the volume of the solids mounds. The mounds were photographed as they were exposed at different tank waste levels to develop a composite of topographical areas. 3. Laser rangefinders to determine the volume of

  10. Bulk crystal growth and nonlinear optical characterization of semiorganic single crystal: Cadmium (II) dibromide L - Proline monohydrate

    Balakrishnan, T., E-mail: [Crystal Growth Laboratory, PG & Research Department of Physics, Periyar EVR College (Autonomous), Tiruchirappalli, 620 023, Tamil Nadu (India); Sathiskumar, S. [Crystal Growth Laboratory, PG & Research Department of Physics, Periyar EVR College (Autonomous), Tiruchirappalli, 620 023, Tamil Nadu (India); Ramamurthi, K. [Crystal Growth and Thin Film Laboratory, Department of Physics and Nanotechnology, SRM University, Kattankulathur, 603 203, Kancheepuram, Tamil Nadu (India); Thamotharan, S. [Department of Bioinformatics, School of Chemical and Biotechnology, SASTRA University, Thanjavur, 613 401 (India)


    Single crystal of a novel metal organic nonlinear optical (NLO) cadmium (II) dibromide L - proline monohydrate (CBLPM) of size 7 × 7 × 5 mm{sup 3} was grown from slow evaporation technique. Single crystal X – ray diffraction analysis reveals that the crystal belongs to orthorhombic system with lattice parameters a = 10.1891 (8) Å, b = 13.4961 (11) Å, c = 7.4491 (5) Å and space group P2{sub 1}2{sub 1}2{sub 1}. The powder X – ray diffraction pattern of CBLPM was recorded and the X – ray diffraction peaks were indexed. The various functional groups of CBLPM were identified by the FT – IR and FT – Raman spectral analyses. The optical transmittance window and lower cut off wavelength of CBLPM were identified from UV – Vis – NIR studies. The mechanical strength of the grown crystal was estimated using Vickers microhardness test. Dielectric constant and dielectric loss measurements were carried out at different temperatures in the frequency range of 50 Hz - 2 MHz. The photoluminescence spectrum was recorded in the wavelength range 200–400 nm and the estimated optical band gap was ∼4.1 eV. Etching studies were carried out for different etching time. Thermal stability of CBLPM was determined using thermogravimetric analysis. Laser induced damage threshold study was carried out for the grown crystal using Nd:YAG laser. Size dependent second harmonic generation efficiency of the grown crystal was determined by Kurtz and Perry powder technique with different particle size using Nd:YAG laser with wavelength 1064 nm. Second harmonic generation efficiency of the powdered CBLPM crystal was ∼2.3 times that of potassium dihydrogen orthophosphate. - Highlights: • CBLPM crystal belongs to orthorhombic crystal system with space group P2{sub 1}2{sub 1}2{sub 1.} • Transmittance of CBLPM is ∼80% in the 650–1100 nm range. • Powder SHG efficiency of CBLPM increases with increase in particle size. • SHG efficiency of 0.57 μm size powdered CBLPM is ∼2

  11. Parameter Estimation


    of optimisation techniques coupled with dynamic solution of the underlying model. Linear and nonlinear approaches to parameter estimation are investigated. There is also the application of maximum likelihood principles in the estimation of parameters, as well as the use of orthogonal collocation to generate a set......In this chapter the importance of parameter estimation in model development is illustrated through various applications related to reaction systems. In particular, rate constants in a reaction system are obtained through parameter estimation methods. These approaches often require the application...... of algebraic equations as the basis for parameter estimation.These approaches are illustrated using estimations of kinetic constants from reaction system models....

  12. Effet de la salinité sur la fluorescence chlorophyllienne, la teneur en proline et la production florale de trois espèces ornementales

    Mathlouthi, M.


    Full Text Available Effect of Chloride Sodium on Chlorophyll Fluorescence, Plant Proline Content and Flowers Production of Three Ornamental Species. Three ornamental species (Zinnia elegans, Tagetes patula and Petunia hybrida were used to test sodium chloride effect on chlorophyll fluorescence, plant proline content and flowers production. Three treatments were used in this trial: 0, 2 and 4 g of Nacl.l-1 of irrigation water. The results showed that chlorophyll fluorescence was not affected by sodium chloride treatment but plant proline content increased and flowers production decreased as NaCl doses increase.

  13. Comparative chlorophyll content and proline metabolism in wheat seedlings exposed to copper and iron stresses%CuCl2和FeCl3处理下小麦幼苗叶绿素含量和脯氨酸代谢的比较

    杨颖丽; 李小宁; 张菁; 乔克雄; 马胜春; 缪建顺; 徐逸菲


    In this study wheat seedlings cv Ningchun 4 is used to evaluate the changes of chlorophyll and osmotic regulator contents as well as the activities of proline metabolic enzymes in response to different CuCl2 and FeCl3 concentrations. The results show that chlorophyll a', chlorophyll b and total chlorophyll contents significantly increas in wheat treated with 100 μmol·L-1 copper, while these three parameters due to 300 μmol'L-1 and 500 μmol·L-1 Cu are obviously lower than those in untreated seedlings. By contrast, the 100 μmol · L-1 concentration of iron do not affect the amount of chlorophyll a, total chlorophyll, soluble sugar and soluble protein, but these parameters significantly elevate in the leaves of wheat under 300 μmol·LT-1 and 500 μmol·L-1 Fe3+ treatments. In addition, Cu2+ and Fe3+ treatments induce an increase in proline content in wheat seedlings, but the enhanced degree under FeCl3 treatment is markedly higher than that in response to CuCl2. Further studies show that both copper and iron treatments result in the decrease of PDH activity, but a notable elevation in the activity of OAT is caused by 500 μmol'L-1 Cu2+ and different Fe3+ concentrations. In conclusion, these results suggest that 300 L-1 and 500 μmol·L-1 CuCl2 treatments lead to damage to chlorophyll in wheat seedlings, while 100 L-1 CuCl2, 300 μmol·L-1 and 500 μmol·L-1 FeCl3 treatments have benefit to the synthesis of chlorophyll; both copper and iron induce the enhancements of soluble sugar, soluble protein and proline content, resulting in strong osmotic adjustment ability, which may improve wheat tolerance to Cu2+ and Fe3+ treatments. Additionally, proline accumulation may be associated with decreased PDH activity and elevated OAT activity in wheat seedlings under copper and iron stresses.%以小麦“宁春四号”为供试材料,研究其在不同浓度的CuCl2和FeCl3处理下叶绿素、渗透性调节物含量及脯氨酸代谢酶活

  14. Arsenate and arsenite exposure modulate antioxidants and amino acids in contrasting arsenic accumulating rice (Oryza sativa L.) genotypes.

    Dave, Richa; Tripathi, Rudra Deo; Dwivedi, Sanjay; Tripathi, Preeti; Dixit, Garima; Sharma, Yogesh Kumar; Trivedi, Prabodh Kumar; Corpas, Francisco J; Barroso, Juan B; Chakrabarty, Debasis


    Carcinogenic arsenic (As) concentrations are found in rice due to irrigation with contaminated groundwater in South-East Asia. The present study evaluates comparative antioxidant property and specific amino acid accumulation in contrasting rice genotypes corresponding to differential As accumulation during arsenate (As(V)) and arsenite (As(III)) exposures. The study was conducted on two contrasting As accumulating rice genotypes selected from 303 genotype accessions, in hydroponic conditions. Maximum As accumulation was up to 1181 μg g(-1) dw in the roots of high As accumulating genotype (HARG), and 89 μg g(-1) dw in low As accumulating genotype (LARG) under As(III) exposures. The inorganic As was correlated more significantly upon exposures to As(III) than As(V). In the presence of As(V) various antioxidant enzymes guiacol peroxidase (GPX), ascorbate peroxidase (APX) and superoxide dismutase (SOD) were highly stimulated in HARG. The stress responsive amino acids proline, cysteine, glycine, glutamic acid and methionine showed higher accumulation in HARG than LARG. A clear correlation was found between stress responsive amino acids, As accumulation and antioxidative response. The comparisons between the contrasting genotypes helped to determine the significance of antioxidants and specific amino acid response to As stress.

  15. Use of cis-[18F] fluoro-proline for assessment of exercise-related collagen synthesis in musculoskeletal connective tissue

    Skovgaard, Dorthe; Kjaer, Andreas; Heinemeier, Katja Maria


    Protein turnover in collagen rich tissue is influenced by exercise, but can only with difficulty be studied in vivo due to use of invasive procedure. The present study was done to investigate the possibility of applying the PET-tracer, cis-[(18)F]fluoro-proline (cis-Fpro), for non-invasive assess......Protein turnover in collagen rich tissue is influenced by exercise, but can only with difficulty be studied in vivo due to use of invasive procedure. The present study was done to investigate the possibility of applying the PET-tracer, cis-[(18)F]fluoro-proline (cis-Fpro), for non......-invasive assessment of collagen synthesis in rat musculoskeletal tissues at rest and following short-term (3 days) treadmill running. Musculoskeletal collagen synthesis was studied in rats at rest and 24 h post-exercise. At each session, rats were PET scanned at two time points following injection of cis-FPro: (60...

  16. The salicylic acid effect on the Salvia officianlis L. sugar, protein and proline contents under salinity (NaCl stress

    Khosravi Sahar


    Full Text Available Plant growth is impressed by biotic and abiotic stress inversely. There are many reports about proteins change level in salinity stress. Leaves fill up more soluble sugar of glucose, fructose and proline with treatment of salicylic acid. In this research, Salivia officialis seeds planted in pots containing perlite were put in a growth chamber under controlled conditions of 27 ±2 0C and 23 ±2 0C temperature, 14h lightness and 10h darkness; NaCl concentration of 0,4,8,12 ds/m and salicylic acid concentration of 0,1,2,4 mM were used in the form of factorial experiment in a complete randomized design (CRD. The results demonstrated that increasing of proline and sugars due to osmotic slope in plants lead to increasing of tolerance against dehydrations of leave content and acceleration of plant developments in stress conditions.

  17. Proline and benzylpenicillin derivatives grafted into mesoporous MCM-41: Novel organic-inorganic hybrid catalysts for direct aldol reaction

    Dwairath Dhar; Ian Beadham; Srinivasan Chandrasekaran


    New organic-inorganic hybrid catalysts were synthesized by covalent grafting of proline and benzylpenicillin derivatives into mesoporous MCM-41. These catalysts were extensively characterized using FT-IR, 13C CP MAS solid state NMR, XRD and TEM techniques. These were used as catalysts for direct, asymmetric aldol reaction between acetone and activated aromatic aldehydes. In the reaction of 4-nitro and 4-fluoro benzaldehyde, the aldol products were obtained in 36% and 59% ee respectively. The catalysts were reusable with neither significant drop in enantioselectivity nor loss of mesostructure. An attempt was made to substantiate the proposed `enamine’ mechanism for direct aldol reaction by trapping the intermediate between proline-MCM-41 and acetone.

  18. A proline-rich polypeptide from ovine colostrum: colostrinin with immunomodulatory activity.

    Zimecki, Michal


    A proline-rich polypeptide (PRP), later called colostrinin (CLN), was originally found as a fraction accompanying sheep colostral immunoglobulins. Extensive in vitro and in vivo studies in mice revealed its interesting T cell-tropic activities. The polypeptide promoted T cell maturation from early thymic precursors that acquired the phenotype and function of mature, helper cells; on the other hand, it also affected the phenotype and function of mature T cells. In particular, PRP was shown to recruit suppressor T cells in a model of T cell-independent humoral immune response and suppressed autoimmune hemolytic anemia in New Zealand Black mice. Subsequent in vitro studies in the human model revealed that CLN regulated mitogen-induced cytokine production in whole blood cultures. A discovery that CLN promoted procognitive functions in experimental animal models, supported by other laboratory findings, indicating prevention of pathological processes in the central nervous system, led to application of CLN in multicenter clinical trials. The trials demonstrated the therapeutic benefit of CLN in Alzheimer's disease (AD) patients by delaying progress of the disease.

  19. Conformational preferences of proline derivatives incorporated into vasopressin analogues: NMR and molecular modelling studies.

    Sikorska, Emilia; Sobolewski, Dariusz; Kwiatkowska, Anna


    In this study, arginine vasopressin analogues modified with proline derivatives - indoline-2-carboxylic acid (Ica), (2S,4R)-4-(naphthalene-2-ylmethyl)pyrrolidine-2-carboxylic acid (Nmp), (2S,4S)-4-aminopyroglutamic acid (APy) and (2R,4S)-4-aminopyroglutamic acid, (Apy) - were examined using NMR spectroscopy and molecular modelling methods. The results have shown that Ica is involved in the formation of the cis peptide bond. Moreover, it reduces to a great extent the conformational flexibility of the peptide. In turn, incorporation of (2S,4R)-Nmp stabilizes the backbone conformation, which is heavily influenced by the pyrrolidine ring. However, the aromatic part of the Nmp side chain exhibits a high degree of conformational freedom. With analogues IV and V, introduction of the 4-aminopyroglumatic acid reduces locally conformational space of the peptides, but it also results in weaker interactions with the dodecylphosphocholine/sodium dodecyl sulphate micelle. Admittedly, both analogues are adsorbed on the micelle's surface but they do not penetrate into its core. With analogue V, the interactions between the peptide and the micelle seem to be so weak that conformational equilibrium is established between different bound states.

  20. The Role of Histidine-Proline-Rich Glycoprotein as Zinc Chaperone for Skeletal Muscle AMP Deaminase

    Maria Ranieri-Raggi


    Full Text Available Metallochaperones function as intracellular shuttles for metal ions. At present, no evidence for the existence of any eukaryotic zinc-chaperone has been provided although metallochaperones could be critical for the physiological functions of Zn2+ metalloenzymes. We propose that the complex formed in skeletal muscle by the Zn2+ metalloenzyme AMP deaminase (AMPD and the metal binding protein histidine-proline-rich glycoprotein (HPRG acts in this manner. HPRG is a major plasma protein. Recent investigations have reported that skeletal muscle cells do not synthesize HPRG but instead actively internalize plasma HPRG. X-ray absorption spectroscopy (XAS performed on fresh preparations of rabbit skeletal muscle AMPD provided evidence for a dinuclear zinc site in the enzyme compatible with a (μ-aqua(μ-carboxylatodizinc(II core with two histidine residues at each metal site. XAS on HPRG isolated from the AMPD complex showed that zinc is bound to the protein in a dinuclear cluster where each Zn2+ ion is coordinated by three histidine and one heavier ligand, likely sulfur from cysteine. We describe the existence in mammalian HPRG of a specific zinc binding site distinct from the His-Pro-rich region. The participation of HPRG in the assembly and maintenance of skeletal muscle AMPD by acting as a zinc chaperone is also demonstrated.

  1. Alzheimer's disease-causing proline substitutions lead to presenilin 1 aggregation and malfunction.

    Ben-Gedalya, Tziona; Moll, Lorna; Bejerano-Sagie, Michal; Frere, Samuel; Cabral, Wayne A; Friedmann-Morvinski, Dinorah; Slutsky, Inna; Burstyn-Cohen, Tal; Marini, Joan C; Cohen, Ehud


    Do different neurodegenerative maladies emanate from the failure of a mutual protein folding mechanism? We have addressed this question by comparing mutational patterns that are linked to the manifestation of distinct neurodegenerative disorders and identified similar neurodegeneration-linked proline substitutions in the prion protein and in presenilin 1 that underlie the development of a prion disorder and of familial Alzheimer's disease (fAD), respectively. These substitutions were found to prevent the endoplasmic reticulum (ER)-resident chaperone, cyclophilin B, from assisting presenilin 1 to fold properly, leading to its aggregation, deposition in the ER, reduction of γ-secretase activity, and impaired mitochondrial distribution and function. Similarly, reduced quantities of the processed, active presenilin 1 were observed in brains of cyclophilin B knockout mice. These discoveries imply that reduced cyclophilin activity contributes to the development of distinct neurodegenerative disorders, propose a novel mechanism for the development of certain fAD cases, and support the emerging theme that this disorder can stem from aberrant presenilin 1 function. This study also points at ER chaperones as targets for the development of counter-neurodegeneration therapies.

  2. Mutagenesis of nisin's leader peptide proline strongly modulates export of precursor nisin.

    Plat, Annechien; Kuipers, Anneke; Crabb, Joe; Rink, Rick; Moll, Gert N


    The lantibiotic nisin is produced by Lactococcus lactis as a precursor peptide comprising a 23 amino acid leader peptide and a 34 amino acid post-translationally modifiable core peptide. We previously demonstrated that the conserved FNLD part of the leader is essential for intracellular enzyme-catalyzed introduction of lanthionines in the core peptide and also for transporter-mediated export, whereas other positions are subject to large mutational freedom. We here demonstrate that, in the absence of the extracellular leader peptidase, NisP, export of precursor nisin via the modification and transporter enzymes, NisBTC, is strongly affected by multiple substitutions of the leader residue at position -2, but not by substitution of positions in the vicinity of this site. Export levels of precursor nisin increased by more than 70% for position -2 mutants Asp, Thr, Ser, Trp, Lys, Val and decreased more than 70% for Cys, His, Met. In a strain with leader peptidase, the Pro-2Lys and Pro-2Asp precursor nisins were less efficiently cleaved by NisP than wild type precursor nisin. Taken together, the wild type precursor nisin with a proline at position -2 allows balanced export and cleavage efficiencies by precursor nisin's transporter and leader peptidase.

  3. The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria.

    Simonyan, Lilit; Légiot, Alexandre; Lascu, Ioan; Durand, Grégory; Giraud, Marie-France; Gonzalez, Cécile; Manon, Stéphen


    Bax is a major player in the apoptotic process, being at the core of the mitochondria permeabilization events. In spite of the major recent advances in the knowledge of Bax organization within the membrane, the precise behavior of the C-terminal helix α9 remains elusive, since it was absent from the resolved structure of active Bax. The Proline 168 (P168) residue, located in the short loop between α8 and α9, has been the target of site-directed mutagenesis experiments, with conflicting results. We have produced and purified a recombinant mutant Bax-P168A, and we have compared its behavior with that of wild-type Bax in a series of tests on Large Unilamellar Vesicles (LUVs) and isolated mitochondria. We conclude that Bax-P168A had a greater ability to oligomerize and bind to membranes. Bax-P168A was not more efficient than wild-type Bax to permeabilize liposomes to small molecules but was more prone to release cytochrome c from mitochondria. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. ITER helium ash accumulation

    Hogan, J.T.; Hillis, D.L.; Galambos, J.; Uckan, N.A. (Oak Ridge National Lab., TN (USA)); Dippel, K.H.; Finken, K.H. (Forschungszentrum Juelich GmbH (Germany, F.R.). Inst. fuer Plasmaphysik); Hulse, R.A.; Budny, R.V. (Princeton Univ., NJ (USA). Plasma Physics Lab.)


    Many studies have shown the importance of the ratio {upsilon}{sub He}/{upsilon}{sub E} in determining the level of He ash accumulation in future reactor systems. Results of the first tokamak He removal experiments have been analysed, and a first estimate of the ratio {upsilon}{sub He}/{upsilon}{sub E} to be expected for future reactor systems has been made. The experiments were carried out for neutral beam heated plasmas in the TEXTOR tokamak, at KFA/Julich. Helium was injected both as a short puff and continuously, and subsequently extracted with the Advanced Limiter Test-II pump limiter. The rate at which the He density decays has been determined with absolutely calibrated charge exchange spectroscopy, and compared with theoretical models, using the Multiple Impurity Species Transport (MIST) code. An analysis of energy confinement has been made with PPPL TRANSP code, to distinguish beam from thermal confinement, especially for low density cases. The ALT-II pump limiter system is found to exhaust the He with maximum exhaust efficiency (8 pumps) of {approximately}8%. We find 1<{upsilon}{sub He}/{upsilon}{sub E}<3.3 for the database of cases analysed to date. Analysis with the ITER TETRA systems code shows that these values would be adequate to achieve the required He concentration with the present ITER divertor He extraction system.

  5. The Binding of Syndapin SH3 Domain to Dynamin Proline-rich Domain Involves Short and Long Distance Elements.

    Luo, Lin; Xue, Jing; Kwan, Ann; Gamsjaeger, Roland; Wielens, Jerome; von Kleist, Lisa; Cubeddu, Liza; Guo, Zhong; Stow, Jennifer L; Parker, Michael W; Mackay, Joel P; Robinson, Phillip J


    Dynamin is a GTPase that mediates vesicle fission during synaptic vesicle endocytosis. Its long C-terminal proline-rich domain contains 13 PXXP motifs, which orchestrate its interactions with multiple proteins. The SH3 domains of syndapin and endophilin bind the PXXP motifs called Site 2 and 3 (Pro-786-Pro-793) at the N-terminal end of the proline-rich domain, whereas the amphiphysin SH3 binds Site 9 (Pro-833-Pro-836) toward the C-terminal end. In some proteins, SH3/peptide interactions also involve short distance elements, which are 5-15 amino acid extensions flanking the central PXXP motif for high affinity binding. Here we found two previously unrecognized elements in the central and the C-terminal end of the dynamin proline-rich domain that account for a significant increase in syndapin binding affinity compared with a previously reported Site 2 and Site 3 PXXP peptide alone. The first new element (Gly-807-Gly-811) is short distance element on the C-terminal side of Site 2 PXXP, which might contact a groove identified under the RT loop of the SH3 domain. The second element (Arg-838-Pro-844) is located about 50 amino acids downstream of Site 2. These two elements provide additional specificity to the syndapin SH3 domain outside of the well described polyproline-binding groove. Thus, the dynamin/syndapin interaction is mediated via a network of multiple contacts outside the core PXXP motif over a previously unrecognized extended region of the proline-rich domain. To our knowledge this is the first example among known SH3 interactions to involve spatially separated and extended long-range elements that combine to provide a higher affinity interaction.

  6. Association of the leucine-7 to proline-7 variation in the signal sequence of neuropeptide Y with major depression

    Koefoed, Pernille; Woldbye, David Paul Drucker; Hansen, Thomas V.O.; Eplov, Lene F; Christiansen, Søren Hofman Oliveira; Mors, Ole; Kessing, Lars Vedel; Werge, Thomas; Kaipio, Katja; Pesonen, Ullamani; Fahmy, Thomas; Mellerup, Erling Thyge; Jakobsen, Klaus D; Hansen, Elsebeth S; Knudsen, Gitte Moos


    Objective: There is clear evidence of a genetic component in major depression, and several studies indicate that neuropeptide Y (NPY) could play an important role in the pathophysiology of the disease. A well-known polymorphism encoding the substitution of leucine to proline in the signal peptide sequence of NPY (Leu7Pro variation) was previously found to protect against depression. Our study aimed at replicating this association in a large Danish population with major depression. Method: Leu...

  7. Stereocontrol in proline-catalyzed asymmetric amination: a comparative assessment of the role of enamine carboxylic acid and enamine carboxylate.

    Sharma, Akhilesh K; Sunoj, Raghavan B


    The transition state models in two mechanistically distinct pathways, involving (i) an enamine carboxylic acid (path-A, 4) and (ii) an enamine carboxylate (path-B, 8), in the proline-catalyzed asymmetric α-amination have been examined using DFT methods. The path-A predicts the correct product stereochemistry under base-free conditions while path-B accounts for reversal of configuration in the presence of a base.

  8. Structural and functional analysis of the yeast N-acetyltransferase Mpr1 involved in oxidative stress tolerance via proline metabolism.

    Nasuno, Ryo; Hirano, Yoshinori; Itoh, Takafumi; Hakoshima, Toshio; Hibi, Takao; Takagi, Hiroshi


    Mpr1 (sigma1278b gene for proline-analog resistance 1), which was originally isolated as N-acetyltransferase detoxifying the proline analog L-azetidine-2-carboxylate, protects yeast cells from various oxidative stresses. Mpr1 mediates the L-proline and L-arginine metabolism by acetylating L-Δ(1)-pyrroline-5-carboxylate, leading to the L-arginine-dependent production of nitric oxide, which confers oxidative stress tolerance. Mpr1 belongs to the Gcn5-related N-acetyltransferase (GNAT) superfamily, but exhibits poor sequence homology with the GNAT enzymes and unique substrate specificity. Here, we present the X-ray crystal structure of Mpr1 and its complex with the substrate cis-4-hydroxy-L-proline at 1.9 and 2.3 Å resolution, respectively. Mpr1 is folded into α/β-structure with eight-stranded mixed β-sheets and six α-helices. The substrate binds to Asn135 and the backbone amide of Asn172 and Leu173, and the predicted acetyl-CoA-binding site is located near the backbone amide of Phe138 and the side chain of Asn178. Alanine substitution of Asn178, which can interact with the sulfur of acetyl-CoA, caused a large reduction in the apparent kcat value. The replacement of Asn135 led to a remarkable increase in the apparent Km value. These results indicate that Asn178 and Asn135 play an important role in catalysis and substrate recognition, respectively. Such a catalytic mechanism has not been reported in the GNAT proteins. Importantly, the amino acid substitutions in these residues increased the L-Δ(1)-pyrroline-5-carboxylate level in yeast cells exposed to heat stress, indicating that these residues are also crucial for its physiological functions. These studies provide some benefits of Mpr1 applications, such as the breeding of industrial yeasts and the development of antifungal drugs.

  9. Novel proline-hydroxyproline glycopeptides from the dandelion (Taraxacum officinale Wigg.) flowers: de novo sequencing and biological activity.

    Astafieva, Alexandra A; Enyenihi, Atim A; Rogozhin, Eugene A; Kozlov, Sergey A; Grishin, Eugene V; Odintsova, Tatyana I; Zubarev, Roman A; Egorov, Tsezi A


    Two novel homologous peptides named ToHyp1 and ToHyp2 that show no similarity to any known proteins were isolated from Taraxacum officinale Wigg. flowers by multidimensional liquid chromatography. Amino acid and mass spectrometry analyses demonstrated that the peptides have unusual structure: they are cysteine-free, proline-hydroxyproline-rich and post-translationally glycosylated by pentoses, with 5 carbohydrates in ToHyp2 and 10 in ToHyp1. The ToHyp2 peptide with a monoisotopic molecular mass of 4350.3Da was completely sequenced by a combination of Edman degradation and de novo sequencing via top down multistage collision induced dissociation (CID) and higher energy dissociation (HCD) tandem mass spectrometry (MS(n)). ToHyp2 consists of 35 amino acids, contains eighteen proline residues, of which 8 prolines are hydroxylated. The peptide displays antifungal activity and inhibits growth of Gram-positive and Gram-negative bacteria. We further showed that carbohydrate moieties have no significant impact on the peptide structure, but are important for antifungal activity although not absolutely necessary. The deglycosylated ToHyp2 peptide was less active against the susceptible fungus Bipolaris sorokiniana than the native peptide. Unique structural features of the ToHyp2 peptide place it into a new family of plant defense peptides. The discovery of ToHyp peptides in T. officinale flowers expands the repertoire of molecules of plant origin with practical applications. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Multiple gas-phase conformations of proline-containing peptides: is it always cis/trans isomerization?

    Lietz, Christopher B; Chen, Zhengwei; Yun Son, Chang; Pang, Xueqin; Cui, Qiang; Li, Lingjun


    Ion mobility-mass spectrometry (IM-MS) is often employed to look at the secondary, tertiary, and quaternary structures of naked peptides and proteins in the gas-phase. Recently, it has offered a unique glimpse into proline-containing peptides and their cis/trans Xxx-Pro isomers. An experimental "signature" has been identified wherein a proline-containing peptide has its Pro residues substituted with another amino acid and the presence or absence of conformations in the IM-MS spectra is observed. Despite the high probability that one could attribute these conformations to cis/trans isomers, it is also possible that cis/trans isomers are not the cause of the additional conformations in proline-containing peptides. However, the experimental evidence of such a system has not been demonstrated or reported. Herein, we present the IM-MS analysis of Neuropeptide Y's wild-type (WT) signal sequence and Leu7Pro (L7P) mutant. Although comparison of arrival times and collision cross-sections of [M + 4H](4+) ions yields the cis/trans "signature", molecular dynamics indicates that a cis-Pro7 is not very stable and that trans-Pro7 conformations of the same cross-section arise with equal frequency. We believe that this work further underscores the importance of theoretical calculations in IM-MS structural assignments.

  11. A Conserved Proline Triplet in Val-tRNA Synthetase and the Origin of Elongation Factor P

    Agata L. Starosta


    Full Text Available Bacterial ribosomes stall on polyproline stretches and require the elongation factor P (EF-P to relieve the arrest. Yet it remains unclear why evolution has favored the development of EF-P rather than selecting against the occurrence of polyproline stretches in proteins. We have discovered that only a single polyproline stretch is invariant across all domains of life, namely a proline triplet in ValS, the tRNA synthetase, that charges tRNAVal with valine. Here, we show that expression of ValS in vivo and in vitro requires EF-P and demonstrate that the proline triplet located in the active site of ValS is important for efficient charging of tRNAVal with valine and preventing formation of mischarged Thr-tRNAVal as well as efficient growth of E. coli in vivo. We suggest that the critical role of the proline triplet for ValS activity may explain why bacterial cells coevolved the EF-P rescue system.

  12. Refined transition-state models for proline-catalyzed asymmetric Michael reactions under basic and base-free conditions.

    Sharma, Akhilesh K; Sunoj, Raghavan B


    The stereocontrolling transition state (TS) models for C-C bond formation relying on hydrogen bonding have generally been successful in proline-catalyzed aldol, Mannich, α-amination, and α-aminoxylation reactions. However, the suitability of the hydrogen-bonding model in protic and aprotic conditions as well as under basic and base-free conditions has not been well established for Michael reactions. Through a comprehensive density functional theory investigation, we herein analyze different TS models for the stereocontrolling C-C bond formation, both in the presence and absence of a base in an aprotic solvent (THF). A refined stereocontrolling TS for the Michael reaction between cyclohexanone and nitrostyrene is proposed. The new TS devoid of hydrogen bonding between the nitro group of nitrostyrene and carboxylic acid of proline, under base-free conditions, is found to be more preferred over the conventional hydrogen-bonding model besides being able to reproduce the experimentally observed stereochemical outcome. A DBU-bound TS is identified as more suitable for rationalizing the origin of asymmetric induction under basic reaction conditions. In both cases, the most preferred approach of nitrostyrene is identified as occurring from the face anti to the carboxylic acid of proline-enamine. The predicted enantio- and diastereoselectivities are in very good agreement with the experimental observations.

  13. Relative Stability of the Key Enamine,Oxazolidinone, and Imine Intermediates in Some Proline-catalyzed Asymmetric Reactions

    HU Yi-Fan; LU Xin


    Many proline-catalyzed asymmetric addition reactions with ketones as substrates were assumed to involve a key intermediate, an enamine, produced by the condensation of proline and ketone. In this paper, the key intermediate enamines derived from L-proline and cyclohexanone (or acetone) as well as the corresponding oxazolidinone and imine tautomers have been investigated by means of density functional calculations at the B3LYP/6-311+G** level. The predicted order of stability for these tautomers is oxazolidinones > enamines > imines in gas phase and oxazolidinones > imines > enamines in aprotic THF solvent. This prediction explains why enamine intermediate can not be observed experimentally. The predicted energy/enthalpy difference between the formal oxazolidinone structure and the zwitterionic imine structures is very small in THF solvent, suggesting the oxazolidinone-to-imine tautomerization can be readily induced in solvent. 13C NMR chemical shifts of the oxazolidinone and imine structures have been computed and used to explain the experimental NMR spectra observed in oxazolidinone-to-imine tautomerization induced by protic solvent.

  14. Modular pathway engineering of Corynebacterium glutamicum for production of the glutamate-derived compounds ornithine, proline, putrescine, citrulline, and arginine.

    Jensen, Jaide V K; Eberhardt, Dorit; Wendisch, Volker F


    The glutamate-derived bioproducts ornithine, citrulline, proline, putrescine, and arginine have applications in the food and feed, cosmetic, pharmaceutical, and chemical industries. Corynebacterium glutamicum is not only an excellent producer of glutamate but also of glutamate-derived products. Here, engineering targets beneficial for ornithine production were identified and the advantage of rationally constructing a platform strain for the production of the amino acids citrulline, proline, and arginine, and the diamine putrescine was demonstrated. Feedback alleviation of N-acetylglutamate kinase, tuning of the promoter of glutamate dehydrogenase gene gdh, lowering expression of phosphoglucoisomerase gene pgi, along with the introduction of a second copy of the arginine biosynthesis operon argCJB(A49V,M54V)D into the chromosome resulted in a C. glutamicum strain producing ornithine with a yield of 0.52 g ornithine per g glucose, an increase of 71% as compared to the parental ΔargFRG strain. Strains capable of producing 0.41 g citrulline per g glucose, 0.29 g proline per g glucose, 0.30 g arginine per g glucose, and 0.17 g putrescine per g glucose were derived from the ornithine-producing platform strain by plasmid-based overexpression of appropriate pathway modules with one to three genes.

  15. Differential contribution of the proline and glutamine pathways to glutamate biosynthesis and nitrogen assimilation in yeast lacking glutamate dehydrogenase.

    Sieg, Alex G; Trotter, Pamela J


    In Saccharomyces cerevisiae, the glutamate dehydrogenase (GDH) enzymes play a pivotal role in glutamate biosynthesis and nitrogen assimilation. It has been proposed that, in GDH-deficient yeast, either the proline utilization (PUT) or the glutamine synthetase-glutamate synthase (GS/GOGAT) pathway serves as the alternative pathway for glutamate production and nitrogen assimilation to the exclusion of the other. Using a gdh-null mutant (gdh1Δ2Δ3Δ), this ambiguity was addressed using a combination of growth studies and pathway-specific enzyme assays on a variety of nitrogen sources (ammonia, glutamine, proline and urea). The GDH-null mutant was viable on all nitrogen sources tested, confirming that alternate pathways for nitrogen assimilation exist in the gdh-null strain. Enzyme assays point to GS/GOGAT as the primary alternative pathway on the preferred nitrogen sources ammonia and glutamine, whereas growth on proline required both the PUT and GS/GOGAT pathways. In contrast, growth on glucose-urea media elicited a decrease in GOGAT activity along with an increase in activity of the PUT pathway specific enzyme Δ(1)-pyrroline-5-carboxylate dehydrogenase (P5CDH). Together, these results suggest the alternative pathway for nitrogen assimilation in strains lacking the preferred GDH-dependent route is nitrogen source dependent and that neither GS/GOGAT nor PUT serves as the sole compensatory pathway.

  16. Advantages of cocrystallization in the field of solid-state pharmaceutical chemistry: L-Proline and MnCl(2).

    Tilborg, Anaëlle; Michaux, Catherine; Norberg, Bernadette; Wouters, Johan


    Cocrystallization (formation of a "cocrystal") is an emerging method to optimize physico-chemical properties of pharmaceutically active compounds. One elegant technique used to obtain such cocrystals is grinding the components together, either alone or in the presence of a small amount of solvent (so called solvent-drop grinding). Dry grinding has been used here to obtain cocrystals (actually a hydrated salt) of L-Proline and MnCl(2). In that context, a new crystalline structure of a multicomponent molecular complex composed of L-Proline and MnCl(2) is here reported. The complex was characterized by powder and single-crystal X-ray diffraction and differential scanning calorimetry. This study underlines the interest of grinding as a method to synthesize original solid-state complexes. It also emphasizes the advantage of combining calorimetric and X-ray diffraction to characterize the newly formed solids. Finally, our work provides structural basis for the role that L-Proline can play within multicomponent solid-state molecular complexes, in particular as a potential cocrystal former acting by both ionic and H-bond interactions when combined to molecules of pharmaceutical interest.

  17. Isolation and characterization of a Sinorhizobium fredii mutant that cannot utilize proline as the sole carbon and nitrogen source

    HUANG Sheng; BAI Xueliang; MA Qingsheng; TANG Xianlai; WU Bo


    Sinorhizobium fredii strain HN01 can use proline as the sole carbon and nitrogen source. A mutant strain GXHN100 unable to catabolize proline was screened from 6000 Tn5gusA5 random insertional mutants of S.fredii strain HN01. Sequencing analysis showed that an open reading frame, named pmrA (proline metabolic relative), was inserted by the Tn5gusA5. A positive clone, named pGXHN100 which containing 3.3kb foreign DNA fragment of S.fredii strain HN01, was isolated from a partial gene library of S.fredii HN01 by colony in situ hybridization. Sequence analysis showed that pGXHN100 contained the entire pmrA gene. The 3.3kb DNA fragment of pGXHN100 was cloned into a broad-host-range cosmid vector pLAFR3 to form plasmid pGXHN200 which was subsequently introduced into GXHN100 to form a complemented strain GXHN200. Plant test showed that GXHN100 was effective and no obvious changes in nitrogenase activity comparing with parental strain. But GXHN100 nodulated 2 days later on soybean and its nodulation efficiency and competitiveness were decreased. The complemented strain GXHN200 restored the nodulation efficiency and competitiveness of GXHN100 to the wild type.

  18. Lipid signalling mediated by PLD/PA modulates proline and H2O2 levels in barley seedlings exposed to short- and long-term chilling stress.

    Peppino Margutti, Micaela; Reyna, Matias; Meringer, María Verónica; Racagni, Graciela E; Villasuso, Ana Laura


    Phospholipase D (PLD) hydrolyses phospholipids to yield phosphatidic acid (PA) and a head group, and is involved in responses to a variety of environmental stresses, including chilling and freezing stress. Barley responses to chilling stress (induced by incubating seedlings at 4 °C) are dynamic and the duration of stress, either short (0-180 min) or long-term (24-36 h) had a significant impact on the response. We investigated the roles of PLD/PA in responses of barley (Hordeum vulgare) seedlings to short and long-term chilling stress, based on regulation of proline and reactive oxygen species (ROS) levels. Short-term chilling stress caused rapid and transient increases in PLD activity, proline level, and ROS levels in young leaves. PLD has the ability to catalyse the transphosphatidylation reaction leading to formation of phosphatidylalcohol (preferentially, to PA). Pre-treatment of seedlings with 1-butanol significantly increased proline synthesis but decreased ROS (H2O2) formation. These observations suggest that PLD is a negative regulator of proline synthesis, whereas PA/PLD promote ROS signals. Exogenous PA pre-treatment reduced the proline synthesis but enhanced H2O2 formation. Effects of long-term chilling stress on barley seedlings differed from those of short-term chilling stress. E.g., PLD activity was significantly reduced in young leaves and roots, whereas proline synthesis and ROS signals were increased in roots. Exogenous ROS application enhanced proline level while exogenous proline application reduced ROS level and modulated some effects of long-term chilling stress. Our findings suggest that PLD contributes to signalling pathways in responses to short-term chilling stress in barley seedling, through regulation of the balance between proline and ROS levels. In contrast, reduced PLD activity in the response to long-term chilling stress did not affect proline level. Increased ROS levels may reflect an antioxidant system that is affected by

  19. 榆钱菠菜脯氨酸转运蛋白基因的克隆及转基因拟南芥的耐盐性%Overexpression of Proline Transporter Gene Isolated from Halophyte Confers Salt Tolerance in Arabidopsis

    沈义国; 张万科; 阎冬青; 杜保兴; 张劲松; 陈受宜


    脯氨酸是自然界中分布最广泛,作用最重要的渗透保护剂之一,同时又是高等植物中一类重要的碳源和氮源物质.为了解环境胁迫下脯氨酸的转运调节,从一个典型的盐生植物榆钱菠菜( Atriplex hortensis L.)中通过cDNA文库筛选和5′-RACE的方法获得了一个全长的cDNA (AhProT1),其编码蛋白与脯氨酸转运蛋白有60%~69%的同源性,含有11个跨膜结构域.聚类分析表明,微生物和高等植物的脯氨酸转运蛋白同源程度相对高于哺乳动物.为进一步分析脯氨酸转运蛋白在植物中的功能,将AhProT1置于35S启动子下转入拟南芥(Arabidopsis thaliana).通过同位素示踪法发现, 与对照植物相比, 转基因植物在根中积累更多的脯氨酸;在一系列不同浓度的盐胁迫试验中,转基因植株最高可耐受200 mmol/L NaCl,并可持续生长,而对照植株在150 mmol/L NaCl下即已死亡.%Proline is one of the most important and widespread osmolyte which functions in adaptation to adverse environmental stresses in many organisms. Also it is an important carbon and nitrogen resource in higher plants. Metabolism of proline has been elucidated in many plant species. However, transport of proline was poorly characterized although transport system plays an important role in proline distribution in different tissues. We isolated one full-length cDNA encoding proline transporter from the typical halophyte: Atriplex hortensis L. through cDNA library screening and 5′-RACE. The deduced amino acid sequence had eleven transmembrane domains, showed 60%-69% similarities to other ProTs and the gene was designated AhProT1. In the phylogenetic tree, higher plants' ProTs, e.g. AhProT1, showed more similar to ProP from microorganisms than ProT from mammalians. AhProT1 gene was transformed into Arabidopsis thaliana under 35S promoter. In MS medium containing [U-14C] proline, AhProT1+ plants were able to accumulate much more radiolabeled proline in the

  20. Articulatory Parameters.

    Ladefoged, Peter


    Summarizes the 16 parameters hypothesized to be necessary and sufficient for linguistic phonetic specifications. Suggests seven parameters affecting tongue shapes, three determining the positions of the lips, one controlling the position of the velum, four varying laryngeal actions, and one controlling respiratory activity. (RL)

  1. Parameter Estimation

    Sales-Cruz, Mauricio; Heitzig, Martina; Cameron, Ian;


    of optimisation techniques coupled with dynamic solution of the underlying model. Linear and nonlinear approaches to parameter estimation are investigated. There is also the application of maximum likelihood principles in the estimation of parameters, as well as the use of orthogonal collocation to generate a set...

  2. Characterisation of the active site of a newly-discovered and potentially significant post-proline cleaving endopeptidase called ZIP using LC-MS

    McMahon, Gillian; Collins, Patrick; O'Connor, Brendan


    There are enzymes that specifically recognise the amino acid proline within peptides and proteins that are called post-proline cleaving enzymes. Many of them are implicated in neurodegenerative disorders and psychiatric diseases. ZIP is a newly-discovered one of these peptidases. In this work, it has been purified from bovine serum and subjected to various analytical studies in order to characterise it. A series of reactions between synthesised peptides and ZIP were carried out in order to el...

  3. Metabolism of sup 3 H- and sup 14 C-labeled glutamate, proline, and alanine in normal and adrenalectomized rats using different sites of tracer administration and sampling

    Said, H.M.; Chenoweth, M.; Dunn, A. (Univ. of Southern California, Los Angeles (USA))


    Alanine, glutamate and proline labeled with 14C and 3H were infused into fasted normal and adrenalectomized rats. Alanine was administered by the A-V mode (arterial administration-venous sampling), and glutamate and proline by both the A-V and V-A (venous administration-arterial sampling) modes. The kinetics of 14C alanine and 14C glutamate differed markedly from those of the tritium-labeled compounds, but there was little difference in the kinetics of 3H and 14C proline. The replacement rate calculated from the A-V mode for glutamate was about half that obtained in the V-A mode, but there was little difference with proline. The masses of the amino acids (total content of amino acids in the body) were calculated from the washout curves of the tritium-labeled compounds after the infusion of tracer was terminated. The masses for the normal rats were 407 mumol/kg for alanine, 578 mumol/kg for glutamate and 296 mumol/kg for proline. The so-called distribution spaces calculated conventionally from total masses and the amino acid concentrations in plasma are much greater than the volume of the body, reflecting the fact that amino acid concentrations in tissues greatly exceed those in plasma. Adrenalectomy markedly affected the kinetics of the three amino acids, and their replacement rates were greatly reduced. The proline and glutamate masses were reduced by at least one half, while that of alanine was unchanged. Adrenalectomy markedly reduced the conversion of proline to glutamate. The hydrocortisone regimen used in this study restored the metabolism of alanine and glutamate to normal, but had no effect on that of proline.

  4. Integrated optical fiber lattice accumulators

    Atherton, Adam F


    Approved for public release; distribution is unlimited. Sigma-delta modulators track a signal by accumulating the error between an input signal and a feedback signal. The accumulated energy is amplitude analyzed by a comparator. The comparator output signal is fed back and subtracted from the input signal. This thesis is primarily concerned with designing accumulators for inclusion in an optical sigma-delta modulator. Fiber lattice structures with optical amplifiers are used to perform the...

  5. Correlation Between Red Edge Characteristic Parameters and Leaf Nitrogen Accumulation and Leaf Area Indices of Processing Tomato%加工番茄红边特征参数与叶片氮积累量和叶面积指数的关系

    黄春燕; 王登伟; 黄鼎程; 马云


    本文分析了加工番茄两品种四氮素水平关键生育时期的冠层红边特征.红边参数与其冠层叶片氮积累量(LNA)和叶面积指数(LAI)的相关性.结果表明:加工番茄的红边曲线具有双峰现象,在开花期、盛花期、坐果期,红边位置(λr)向长波方向移动了2~11 nm波段,红边曲线峰值(Dr)较大,LNA和LAI较高.到红果期红边位置向短波方向移动了5~10 nm波段,红边曲线峰值也较低,LNA和LAI最低,双峰现象在红果期随着施氮量的减少而减弱,说明红边位置的变化可以指示加工番茄生育时期的氮索营养状况.加工番茄2个品种冠层的λr、红边斜率(Dr)和红边面积(SDr)分别与其冠层LNA和LAI的相关性均达到了1%极显著水平,其中里格尔87-5的Dr与LNA的线性相关和SDr与LAI的指数相关最高(rD-LNA=0.8181**,rSD-LAI =0.9136**,n=23,α=1%),说明利用红边参数可以实时快速地获取加工番茄冠层LNA和LAI的信息,可有效监测加工番茄的氮素营养状况.%This paper analyzed of the characteristics of red edge of two varieties of processing tomatoes in four nitrogen fertilization treatments at key growing stage, the correlation between three red edge parameters and their corresponding leaf nitrogen accumulation (LNA) ,leaf area indices (LAI). respectively. The results showed that red edge curve of processing tomato had double peaks; red edge position(Ar) shifted towards long wavelengths range 2~11 nm at its flower stage, full bloom stage and fruit setting stage ,com pared with seedling stage;the peak value of red edge curve (Dr) was higher than the other three stages, and LNA and LAI were also higher.λr shifted towards short wavelengths range 5~10 nm at its red fruit stagesthe peak value of red edge curve reached lower; LNA and LAI were both the lowest .double peaks were weakened with the reduction of nitrogen fertilizer rate at red fruit stage;The shift of the red edge position indicated nitrogen

  6. Root Exudates, Rhizosphere Zn Fractions, and Zn Accumulation of Ryegrass at Different Soil Zn Levels

    XU Wei-Hong; LIU Huai; MA Qi-Fu; XIONG Zhi-Ting


    A glasshouse experiment was conducted using a root-bag technique to study the root exudates, rhizosphere Zn fractions, and Zn concentrations and accumulations of two ryegrass cultivars (Lolium perenne L. cvs. Airs and Tede) at different soil Zn levels (0, 2, 4, 8, and 16 mmol kg-1 soil). Results indicated that plant growth of the two cultivars was not adversely affected at soil Zn level≤8 mmol kg-1. Plants accumulated more Zn as soil Zn levels increased, and Zn concentrations of shoots were about 540 /μg g-1 in Aris and 583.9μg g-1 in Tede in response to 16 mmol Zn kg-1 soil. Zn ratios of shoots to roots across the soil Zn levels were higher in Tede than in Airs, corresponding with higher rhizosphere available Zn fractions (exchangeable, bound to manganese oxides, and bound to organic matter) in Airs than in Tede. Low-molecular-weight (LMW) organic acids (oxalic, tartaric, malic, and succinic acids) and amino acids (proline, threonine, glutamic acid, and aspartic acid, etc.) were detected in root exudates, and the concentrations of LMW organic acids and amino acids increased with addition of 4 mmol Zn kg-1 soil compared with zero Zn addition. Higher rhizosphere concentrations of oxalic acid, glutamic acid, alanine, phenylalanine, leucine, and proline in Tede than in Airs likely resulted in increased Zn uptake from the soil by Tede than by Airs. The results suggested that genotypic differences in Zn accumulations were mainly because of different root exudates and rhizosphere Zn fractions.

  7. Assignment of PolyProline II conformation and analysis of sequence--structure relationship.

    Mansiaux, Yohann; Joseph, Agnel Praveen; Gelly, Jean-Christophe; de Brevern, Alexandre G


    Secondary structures are elements of great importance in structural biology, biochemistry and bioinformatics. They are broadly composed of two repetitive structures namely α-helices and β-sheets, apart from turns, and the rest is associated to coil. These repetitive secondary structures have specific and conserved biophysical and geometric properties. PolyProline II (PPII) helix is yet another interesting repetitive structure which is less frequent and not usually associated with stabilizing interactions. Recent studies have shown that PPII frequency is higher than expected, and they could have an important role in protein-protein interactions. A major factor that limits the study of PPII is that its assignment cannot be carried out with the most commonly used secondary structure assignment methods (SSAMs). The purpose of this work is to propose a PPII assignment methodology that can be defined in the frame of DSSP secondary structure assignment. Considering the ambiguity in PPII assignments by different methods, a consensus assignment strategy was utilized. To define the most consensual rule of PPII assignment, three SSAMs that can assign PPII, were compared and analyzed. The assignment rule was defined to have a maximum coverage of all assignments made by these SSAMs. Not many constraints were added to the assignment and only PPII helices of at least 2 residues length are defined. The simple rules designed in this study for characterizing PPII conformation, lead to the assignment of 5% of all amino as PPII. Sequence-structure relationships associated with PPII, defined by the different SSAMs, underline few striking differences. A specific study of amino acid preferences in their N and C-cap regions was carried out as their solvent accessibility and contact patterns. Thus the assignment of PPII can be coupled with DSSP and thus opens a simple way for further analysis in this field.

  8. Assignment of PolyProline II Conformation and Analysis of Sequence – Structure Relationship

    Gelly, Jean-Christophe; de Brevern, Alexandre G.


    Background Secondary structures are elements of great importance in structural biology, biochemistry and bioinformatics. They are broadly composed of two repetitive structures namely α-helices and β-sheets, apart from turns, and the rest is associated to coil. These repetitive secondary structures have specific and conserved biophysical and geometric properties. PolyProline II (PPII) helix is yet another interesting repetitive structure which is less frequent and not usually associated with stabilizing interactions. Recent studies have shown that PPII frequency is higher than expected, and they could have an important role in protein – protein interactions. Methodology/Principal Findings A major factor that limits the study of PPII is that its assignment cannot be carried out with the most commonly used secondary structure assignment methods (SSAMs). The purpose of this work is to propose a PPII assignment methodology that can be defined in the frame of DSSP secondary structure assignment. Considering the ambiguity in PPII assignments by different methods, a consensus assignment strategy was utilized. To define the most consensual rule of PPII assignment, three SSAMs that can assign PPII, were compared and analyzed. The assignment rule was defined to have a maximum coverage of all assignments made by these SSAMs. Not many constraints were added to the assignment and only PPII helices of at least 2 residues length are defined. Conclusions/Significance The simple rules designed in this study for characterizing PPII conformation, lead to the assignment of 5% of all amino as PPII. Sequence – structure relationships associated with PPII, defined by the different SSAMs, underline few striking differences. A specific study of amino acid preferences in their N and C-cap regions was carried out as their solvent accessibility and contact patterns. Thus the assignment of PPII can be coupled with DSSP and thus opens a simple way for further analysis in this field. PMID

  9. Alanine and proline content modulate global sensitivity to discrete perturbations in disordered proteins.

    Perez, Romel B; Tischer, Alexander; Auton, Matthew; Whitten, Steven T


    Molecular transduction of biological signals is understood primarily in terms of the cooperative structural transitions of protein macromolecules, providing a mechanism through which discrete local structure perturbations affect global macromolecular properties. The recognition that proteins lacking tertiary stability, commonly referred to as intrinsically disordered proteins (IDPs), mediate key signaling pathways suggests that protein structures without cooperative intramolecular interactions may also have the ability to couple local and global structure changes. Presented here are results from experiments that measured and tested the ability of disordered proteins to couple local changes in structure to global changes in structure. Using the intrinsically disordered N-terminal region of the p53 protein as an experimental model, a set of proline (PRO) and alanine (ALA) to glycine (GLY) substitution variants were designed to modulate backbone conformational propensities without introducing non-native intramolecular interactions. The hydrodynamic radius (R(h)) was used to monitor changes in global structure. Circular dichroism spectroscopy showed that the GLY substitutions decreased polyproline II (PP(II)) propensities relative to the wild type, as expected, and fluorescence methods indicated that substitution-induced changes in R(h) were not associated with folding. The experiments showed that changes in local PP(II) structure cause changes in R(h) that are variable and that depend on the intrinsic chain propensities of PRO and ALA residues, demonstrating a mechanism for coupling local and global structure changes. Molecular simulations that model our results were used to extend the analysis to other proteins and illustrate the generality of the observed PRO and alanine effects on the structures of IDPs. © 2014 Wiley Periodicals, Inc.

  10. Tyrosine Nitration within the Proline-Rich Region of Tau in Alzheimer's Disease

    Reyes, Juan F.; Fu, Yifan; Vana, Laurel; Kanaan, Nicholas M.; Binder, Lester I.


    A substantial body of evidence suggests that nitrative injury contributes to neurodegeneration in Alzheimer's disease (AD) and other neurodegenerative disorders. Previously, we showed in vitro that within the tau protein the N-terminal tyrosine residues (Y18 and Y29) are more susceptible to nitrative modifications than other tyrosine sites (Y197 and Y394). Using site-specific antibodies to nitrated tau at Y18 and Y29, we identified tau nitrated in both glial (Y18) and neuronal (Y29) tau pathologies. In this study, we report the characterization of two novel monoclonal antibodies, Tau-nY197 and Tau-nY394, recognizing tau nitrated at Y197 and Y394, respectively. By Western blot analysis, Tau-nY197 labeled soluble tau and insoluble paired helical filament proteins (PHF-tau) nitrated at Y197 from control and AD brain samples. Tau-nY394 failed to label soluble tau isolated from control or severe AD samples, but labeled insoluble PHF-tau to a limited extent. Immunohistochemical analysis using Tau-nY197 revealed the hallmark tau pathology associated with AD; Tau-nY394 did not detect any pathological lesions characteristic of the disorder. These data suggest that a subset of the hallmark pathological inclusions of AD contain tau nitrated at Y197. However, nitration at Y197 was also identified in soluble tau from all control samples, including those at Braak stage 0, suggesting that nitration at this site in the proline-rich region of tau may have normal biological functions in the human brain. PMID:21514440

  11. Expression of proline-rich Akt-substrate PRAS40 in cell survival pathway and carcinogenesis

    Bei HUANG; Gavin PORTER


    Aim: To study the expression of proline-rich Akt-substrate PRAS40 in the cell survival pathway and tumor progression. Methods: The effects of three key kinase inhibitors on PRAS40 activity in the cell survival pathway, serum withdrawal,H2O2 and overexpression of Akt were tested. The expression of PRAS40, Akt, Raf and 14-3-3 in normal cells and cancer cell lines was determined by Western blot.Results: The PI3K inhibitors worthmannin and Ly294002, but not rapamycin, completely inhibited the phosphorylation of Akt and PRAS40. The phosphorylation level of Akt decreased after serum withdrawal and treatment with the MEK inhibitor Uo126, but increased after treatment with H2O2 at low concentration, whereas none of these treatments changed PRAS40 activity. 14-3-3 is a PRAS40 binding protein, and the expression of 14-3-3, like that of PRAS40, was higher in HeLa cells than in HEK293 cells; PRAS40 had a stronger phosphorylation activity in A549 and HeLa cancer cells than in HEK293 normal cells. In the breast cancer model (MCF10A/MCF7) and lung cancer model (BEAS/H1198/H1170) we also found the same result: PRAS40 was constitutively active in H1198/H1170 and MCF7 premalignant and malignant cancer cells, but weakly expressed in MCF10A and BEAS normal cell. We also discussed PRAS40 activity in other NSCLC cell lines.Conclusion: The PI3K-Akt survival pathway is the main pathway that PRAS40 is involved in; PRAS40 is a substrate for Akt, but can also be activated by an Aktindependent mechanisms. PRAS40 activation is an early event during breast and lung carcinogenesis.

  12. Activating mutations of the TRPML1 channel revealed by proline-scanning mutagenesis.

    Dong, Xian-ping; Wang, Xiang; Shen, Dongbiao; Chen, Su; Liu, Meiling; Wang, Yanbin; Mills, Eric; Cheng, Xiping; Delling, Markus; Xu, Haoxing


    The mucolipin TRP (TRPML) proteins are a family of endolysosomal cation channels with genetically established importance in humans and rodent. Mutations of human TRPML1 cause type IV mucolipidosis, a devastating pediatric neurodegenerative disease. Our recent electrophysiological studies revealed that, although a TRPML1-mediated current can only be recorded in late endosome and lysosome (LEL) using the lysosome patch clamp technique, a proline substitution in TRPML1 (TRPML1(V432P)) results in a large whole cell current. Thus, it remains unknown whether the large TRPML1(V432P)-mediated current results from an increased surface expression (trafficking), elevated channel activity (gating), or both. Here we performed systemic Pro substitutions in a region previously implicated in the gating of various 6 transmembrane cation channels. We found that several Pro substitutions displayed gain-of-function (GOF) constitutive activities at both the plasma membrane (PM) and endolysosomal membranes. Although wild-type TRPML1 and non-GOF Pro substitutions localized exclusively in LEL and were barely detectable in the PM, the GOF mutations with high constitutive activities were not restricted to LEL compartments, and most significantly, exhibited significant surface expression. Because lysosomal exocytosis is Ca(2+)-dependent, constitutive Ca(2+) permeability due to Pro substitutions may have resulted in stimulus-independent intralysosomal Ca(2+) release, hence the surface expression and whole cell current of TRPML1. Indeed, surface staining of lysosome-associated membrane protein-1 (Lamp-1) was dramatically increased in cells expressing GOF TRPML1 channels. We conclude that TRPML1 is an inwardly rectifying, proton-impermeable, Ca(2+) and Fe(2+)/Mn(2+) dually permeable cation channel that may be gated by unidentified cellular mechanisms through a conformational change in the cytoplasmic face of the transmembrane 5 (TM5). Furthermore, activation of TRPML1 in LEL may lead to the

  13. Mutagenesis of bacteriophage IKe major coat protein transmembrane domain: role of an interfacial proline residue.

    Williams, K A; Deber, C M


    The transmembrane (TM) domain of the 53-residue major coat protein of the M13-related bacteriophage IKe (residues 24-42: LISQTWPVVTTVVVAGVLI) has been subjected to randomized mutagenesis to probe the conformation and stability of the TM domain, as well as the effect of structurally-important residues such as proline. TM mutants were obtained by the Eckstein method of site-directed mutagenesis using the IKe genome as template so as to eliminate the need for subcloning. Over 40 single- and double-site viable mutants of bacteriophage IKe were isolated. Every residue in the TM segment, except the highly conserved Trp29, could be mutated to at least one other residue; polar and charged mutations occurred in the TM segment adjacent to the N-terminal domain (residues 24-28), while non-polar substitutions predominated in the C-terminal portion (residues 30-42). The Pro30 locus tolerated four mutations-Ala, Gly, Cys, and Ser- which represent the four side chains of least volume. Mutant coat proteins obtained directly from the phage in milligram quantities were studied by circular dichroism spectroscopy and SDS-PAGE gels. Wild type IKe coat protein solubilized in sodium deoxycholate micelles was found to occur as an alpha-helical, monomeric species which is stable at 95 degrees C, whereas the mutant Pro30-->Gly undergoes an irreversible conformational transition at ca. 90 degrees C to an aggregated beta-sheet structure. The result that Pro30 stabilizes the TM helix in the micellar membrane suggests a sterically-restricted location for the wild type Pro pyrrolidine side chain in the bulky Trp-Pro-Val triad, where it may be positioned to direct the initiation of the subsequent TM core domain helix.

  14. Organic photodiodes from homochiral l-proline derived squaraine compounds with strong circular dichroism.

    Schulz, Matthias; Mack, Majvor; Kolloge, Oliver; Lützen, Arne; Schiek, Manuela


    We suggest and explore a novel route towards organic photodetectors sensitive to the circular polarization state of light. For this, we insert fullerene-blended thin films of homochiral squaraine compounds acting as a highly circular dichroic active layer into conventional bulk hetero-junction photodiodes. Initially, we discuss steady-state characterization of photodiodes with unpolarized light. The homochiral, l-proline derived squaraine compounds are obtained via a chiral pool synthesis in sizable quantities. The aggregation behavior of the two compounds with varying side chain length is complex. They exhibit H-type spectral signatures only in colloidal solution, and both H- and J-type features with large splitting in neat and fullerene-blended thin films. We probe strong excitonic circular dichroism for both aggregate species, showing the most competitive dissymmetry factors up to -0.055 in fullerene-blended thin films. We vary the blend ratio and layer thickness of such active layers in the photodiodes. The device performance is in all cases limited by a low fill factor which is accompanied by a voltage-dependent photocurrent. Current-voltage measurements show light intensity dependent characteristics, which are S-shaped, contrary to our expectations, only for thin active layers independent of the blend ratio. The external quantum efficiency is in some cases extraordinarily high, exceeding 100 percent in the blue spectral range under modest reverse bias voltages for thin, fullerene-rich devices. However, the most promising are the devices with thick, donor-rich layers defined by a spectral overlap of the strongest photocurrent response and the maximum circular dichroism within the green spectral range. Thus, we demonstrate the feasibility of combining photodiode functionality and strong circular dichroism as intrinsic material properties.

  15. Slow peptide bond formation by proline and other N-alkylamino acids in translation

    Pavlov, Michael Y.; Watts, Richard E.; Tan, Zhongping; Cornish, Virginia W.; Ehrenberg, Måns; Forster, Anthony C.


    Proteins are made from 19 aa and, curiously, one N-alkylamino acid (“imino acid”), proline (Pro). Pro is thought to be incorporated by the translation apparatus at the same rate as the 19 aa, even though the alkyl group in Pro resides directly on the nitrogen nucleophile involved in peptide bond formation. Here, by combining quench-flow kinetics and charging of tRNAs with cognate and noncognate amino acids, we find that Pro incorporates in translation significantly more slowly than Phe or Ala and that other N-alkylamino acids incorporate much more slowly. Our results show that the slowest step in incorporation of N-alkylamino acids is accommodation/peptidyl transfer after GTP hydrolysis on EF-Tu. The relative incorporation rates correlate with expectations from organic chemistry, suggesting that amino acid sterics and basicities affect translation rates at the peptidyl transfer step. Cognate isoacceptor tRNAs speed Pro incorporation to rates compatible with in vivo, although still 3–6 times slower than Phe incorporation from Phe-tRNAPhe depending on the Pro codon. Results suggest that Pro is the only N-alkylamino acid in the genetic code because it has a privileged cyclic structure that is more reactive than other N-alkylamino acids. Our data on the variation of the rate of incorporation of Pro from native Pro-tRNAPro isoacceptors at 4 different Pro codons help explain codon bias not accounted for by the “tRNA abundance” hypothesis. PMID:19104062

  16. Inventory parameters

    Sharma, Sanjay


    This book provides a detailed overview of various parameters/factors involved in inventory analysis. It especially focuses on the assessment and modeling of basic inventory parameters, namely demand, procurement cost, cycle time, ordering cost, inventory carrying cost, inventory stock, stock out level, and stock out cost. In the context of economic lot size, it provides equations related to the optimum values. It also discusses why the optimum lot size and optimum total relevant cost are considered to be key decision variables, and uses numerous examples to explain each of these inventory parameters separately. Lastly, it provides detailed information on parameter estimation for different sectors/products. Written in a simple and lucid style, it offers a valuable resource for a broad readership, especially Master of Business Administration (MBA) students.

  17. The role of Δ1-pyrroline-5-carboxylate dehydrogenase in proline degradation

    Deuschle, Karen; Funck, Dietmar; Forlani, Giuseppe


    In response to stress, plants accumulate Pro, requiring degradation after release from adverse conditions. Delta1-Pyrroline-5-carboxylate dehydrogenase (P5CDH), the second enzyme for Pro degradation, is encoded by a single gene expressed ubiquitously. To study the physiological function of P5CDH,...

  18. Involvement of Phosphatidylinositol 3-kinase in the regulation of proline catabolism in Arabidopsis thaliana

    A.S. Leprince; N. Magalhaes; D. de Vos; M. Bordenave; E. Crilat; G. Clément; C Meyer; T. Munnik; A. Savouré


    Plant adaptation to abiotic stresses such as drought and salinity involves complex regulatory processes. Deciphering the signaling components that are involved in stress signal transduction and cellular responses is of importance to understand how plants cope with salt stress. Accumulation of osmoly

  19. Expression and effect of proline hydroxylase domain 2 in retina of diabetic rats

    Zhen Li


    Full Text Available AIM: To observe the expression of proline hydroxylase domain 2 (PHD2 in the retina of diabetic rats and investigate the relationship between PHD2 and relevant intraocular vascular proliferation factors. METHODS: Sixty male specific pathogen free (SPF Sprague-Dawley (SD rats were randomly divided into two groups: the diabetic group and the control group. The rats in the diabetic group were intraperitoneally injected with 60 mg/kg (0.60 mL/100g of streptozotocin to induce a diabetic rat model. The rats in the control group were injected with an equal volume of sodium citrate buffer solution by the same method. Hematoxylin-eosin (HE staining and immumofluorescence (IF method were adopted to observe the pathological changes of retinal tissues and the expression of PHD2, glial fibrillary acidic protein (GFAP, vascular endothelial growth factor (VEGF by 8wk. RT-PCR method was applied to detect the expressions of mRNA of PHD2, VEGF and GFAP. The relationship between PHD2 and other vascular proliferation factors was analyzed. RESULTS: HE staining showed that there was the retinal tissue edema in the diabetic group, and the arrangement was in disorder, and proliferation could be seen. IF staining: in the retina of normal rats, PHD2 was not expressed, GFAP and VEGF were mainly expressed in astrocytes; while in the diabetic rats, PHD2, GFAP and VEGF staining showed strong positivity in all retinal layers, mainly in neurogliocytes. PHD2 was co-expressed with VEGF and GFAP. The mRNA expression levels of PHD2, GFAP and VEGF in the diabetic group were obviously higher than that in the control group,respectively 1.83 times, 1.75 times and 2.08 times. The difference had statistical significance (P<0.01. CONCLUSION: The high expression of PHD2 in the retina of early-stage diabetic rats might result from secretion of neurogliocytes induced by local high-concentration blood glucose, thus promoting the expression of VEGF and GFAP. PHD2 plays an important role during

  20. Thermostability Improvement of a Streptomyces Xylanase by Introducing Proline and Glutamic Acid Residues

    Wang, Kun; Tian, Jian; Turunen, Ossi; Huang, Huoqing; Shi, Pengjun; Hua, Huifang; Wang, Caihong; Wang, Shuanghe


    Protein engineering is commonly used to improve the robustness of enzymes for activity and stability at high temperatures. In this study, we identified four residues expected to affect the thermostability of Streptomyces sp. strain S9 xylanase XynAS9 through multiple-sequence analysis (MSA) and molecular dynamic simulations (MDS). Site-directed mutagenesis was employed to construct five mutants by replacing these residues with proline or glutamic acid (V81P, G82E, V81P/G82E, D185P/S186E, and V81P/G82E/D185P/S186E), and the mutant and wild-type enzymes were expressed in Pichia pastoris. Compared to the wild-type XynAS9, all five mutant enzymes showed improved thermal properties. The activity and stability assays, including circular dichroism and differential scanning calorimetry, showed that the mutations at positions 81 and 82 increased the thermal performance more than the mutations at positions 185 and 186. The mutants with combined substitutions (V81P/G82E and V81P/G82E/D185P/S186E) showed the most pronounced shifts in temperature optima, about 17°C upward, and their half-lives for thermal inactivation at 70°C and melting temperatures were increased by >9 times and approximately 7.0°C, respectively. The mutation combination of V81P and G82E in adjacent positions more than doubled the effect of single mutations. Both mutation regions were at the end of long secondary-structure elements and probably rigidified the local structure. MDS indicated that a long loop region after positions 81 and 82 located at the end of the inner β-barrel was prone to unfold. The rigidified main chain and filling of a groove by the mutations on the bottom of the active site canyon may stabilize the mutants and thus improve their thermostability. PMID:24463976

  1. Proline-induced hinges in transmembrane helices: possible roles in ion channel gating.

    Tieleman, D P; Shrivastava, I H; Ulmschneider, M R; Sansom, M S


    A number of ion channels contain transmembrane (TM) alpha-helices that contain proline-induced molecular hinges. These TM helices include the channel-forming peptide alamethicin (Alm), the S6 helix from voltage-gated potassium (Kv) channels, and the D5 helix from voltage-gated chloride (CLC) channels. For both Alm and KvS6, experimental data implicate hinge-bending motions of the helix in an aspect of channel gating. We have compared the hinge-bending motions of these TM helices in bilayer-like environments by multi-nanosecond MD simulations in an attempt to describe motions of these helices that may underlie possible modes of channel gating. Alm is an alpha-helical channel-forming peptide, which contains a central kink associated with a Gly-x-x-Pro motif in its sequence. Simulations of Alm in a TM orientation for 10 ns in an octane slab indicate that the Gly-x-x-Pro motif acts as a molecular hinge. The S6 helix from Shaker Kv channels contains a Pro-Val-Pro motif. Modeling studies and recent experimental data suggest that the KvS6 helix may be kinked in the vicinity of this motif. Simulations (10 ns) of an isolated KvS6 helix in an octane slab and in a POPC bilayer reveal hinge-bending motions. A pattern-matching approach was used to search for possible hinge-bending motifs in the TM helices of other ion channel proteins. This uncovered a conserved Gly-x-Pro motif in TM helix D5 of CLC channels. MD simulations of a model of hCLC1-D5 spanning an octane slab suggest that this channel also contains a TM helix that undergoes hinge-bending motion. In conclusion, our simulations suggest a model in which hinge-bending motions of TM helices may play a functional role in the gating mechanisms of several different families of ion channels.

  2. 24-Epibrassinolide regulates photosynthesis, antioxidant enzyme activities and proline content of Cucumis sativus under salt and/or copper stress.

    Fariduddin, Q; Khalil, Radwan R A E; Mir, Bilal A; Yusuf, M; Ahmad, A


    Brassinosteroids have been extensively used to overcome various abiotic stresses. But its role in combined stress of salt and excess copper remains unexplored. Seeds of two cultivars (Rocket and Jumbo) of Cucumis sativus were grown in sand amended with copper (100 mg kg(-1)), and developed seedlings were exposed to salt stress in the form of NaCl (150 mM) at the 30-day stage of growth for 3 days. These seedlings were subsequently sprayed with 0 or 0.01 μM of 24-epibrassinolide (EBL) at the 35-day stage. The plants exposed to NaCl and Cu in combination exhibited a significant decline in fresh and dry mass of plant, chlorophyll content, activities of carbonic anhydrase, net photosynthetic rate and maximum quantum yield of the PSII primary photochemistry followed by NaCl and Cu stress alone, more severely in Jumbo than in Rocket. However, the follow-up treatment with EBL to the stressed and nonstressed plant improved growth, chlorophyll content, carbonic anhydrase activity and photosynthetic efficiency, and further enhanced the activity of various antioxidant enzymes viz. catalase, peroxidase and superoxide dismutase and content of proline at the 40-day stage of growth, and the response of the hormone was more effective in Rocket than in Jumbo. The elevated level of antioxidant enzymes as well as proline could have conferred tolerance to the NaCl- and/or Cu-stressed plants resulting in improved growth, water relations and photosynthetic attributes. Furthermore, antioxidant enzyme activity and proline content were more enhanced in Rocket than in Jumbo cultivar.

  3. Rapid degradation of D- and L-succinimide-containing peptides by a post-proline endopeptidase from human erythrocytes

    Momand, J.; Clarke, S.


    The authors have been interested in the metabolic fate of proteins containing aspartyl succinimide (Asu) residues. These residues can be derived from the spontaneous rearrangement of Asp and Asn residues and from the spontaneous demethylation of enzymatically methylated L-isoAsp and D-Asp residues. Incubation of the synthetic hexapeptide N-Ac-Val-Tyr-Pro-Asu-Gly-Ala with the cytosolic fraction of human erythrocytes resulted in rapid cleavage of the prolyl-aspartyl succinimide bond producing the tripeptide N-Ac-Val-Try-Pro. The rate of this reaction is equal for both L- and D-Asu-containing peptides and is 10-fold greater that the rate of cleavage of a corresponding peptide containing a normal Pro-Asp linkage. When the aspartyl succinimide ring was replaced with an isoaspartyl residue, the cleavage rate was about 5 times that of the normal Pro-Asp peptide. The tripeptide-producing activity copurified on DEAE-cellulose chromatography with an activity that cleaves N-carbobenzoxy-Gly-Pro-4-methylcoumarin-7-amide, a post-proline endopeptidase substrate. These two activities were both inhibited by an antiserum to rat brain post-proline endopeptidase, and it appears that they are catalyzed by the same enzyme. This enzyme has a molecular weight of approximately 80,000 and is covalently labeled and inhibited by (/sup 3/H) diisopropyl fluorophosphate. The facile cleavage of the succinimide- and isoaspartyl-containing peptides by this post-proline endopeptidase suggests that it may play a role in the metabolism of peptides containing altered aspartyl residues.

  4. Biochemical characterization, homology modeling and docking studies of ornithine delta-aminotransferase--an important enzyme in proline biosynthesis of plants.

    Sekhar, P Nataraj; Amrutha, R Naga; Sangam, Shubhada; Verma, D P S; Kishor, P B Kavi


    Ornithine delta-aminotransferase (OAT) is an important enzyme in proline biosynthetic pathway and is implicated in salt tolerance in higher plants. OAT transaminates ornithine to pyrroline 5-carboxylate, which is further catalyzed to proline by pyrroline 5-carboxylate reductase. The Vigna aconitifolia OAT cDNA, encoding a polypeptide of 48.1 kDa, was expressed in Escherichia coli and the enzyme was partially characterized following its purification using (NH(4))(2)SO(4) precipitation and gel filtration techniques. Optimal activity of the enzyme was observed at a temperature of 25 degrees C and pH 8.0. The enzyme appeared to be a monomer and exhibited high activity at 4mM ornithine. Proline did not show any apparent effect but isoleucine, valine and serine inhibited the activity when added into the assay mixture along with ornithine. Omission of pyridoxal 5'-phosphate from the reaction mixture reduced the activity of this enzyme by 60%. To further evaluate these biochemical observations, homology modeling of the OAT was performed based on the crystal structure of the ornithine delta-aminotransferase from humans (PDB code 1OAT) by using the software MODELLER6v2. With the aid of the molecular mechanics and dynamics methods, the final model was obtained and assessed subsequently by PROCHECK and VERIFY-3D graph. With this model, a flexible docking study with the substrate and inhibitors was performed and the results indicated that Gly106 and Lys256 in OAT are the important determinant residues in binding as they have strong hydrogen bonding contacts with the substrate and inhibitors. These observations are in conformity with the results obtained from experimental investigations.

  5. How wealth accumulation can promote cooperation.

    Thomas Chadefaux

    Full Text Available Explaining the emergence and stability of cooperation has been a central challenge in biology, economics and sociology. Unfortunately, the mechanisms known to promote it either require elaborate strategies or hold only under restrictive conditions. Here, we report the emergence, survival, and frequent domination of cooperation in a world characterized by selfishness and a strong temptation to defect, when individuals can accumulate wealth. In particular, we study games with local adaptation such as the prisoner's dilemma, to which we add heterogeneity in payoffs. In our model, agents accumulate wealth and invest some of it in their interactions. The larger the investment, the more can potentially be gained or lost, so that present gains affect future payoffs. We find that cooperation survives for a far wider range of parameters than without wealth accumulation and, even more strikingly, that it often dominates defection. This is in stark contrast to the traditional evolutionary prisoner's dilemma in particular, in which cooperation rarely survives and almost never thrives. With the inequality we introduce, on the contrary, cooperators do better than defectors, even without any strategic behavior or exogenously imposed strategies. These results have important consequences for our understanding of the type of social and economic arrangements that are optimal and efficient.

  6. Manganese As a Metal Accumulator

    Manganese deposits in water distribution systems accumulate metals, radionuclides and oxyanions by a combination of surface complexation, adsorption and solid substitution, as well as a combination of oxidation followed by manganese reduction and sorption of the oxidized constitu...

  7. Manganese As a Metal Accumulator

    Manganese deposits in water distribution systems accumulate metals, radionuclides and oxyanions by a combination of surface complexation, adsorption and solid substitution, as well as a combination of oxidation followed by manganese reduction and sorption of the oxidized constitu...

  8. Influence of Hofmeister Ions on the Structure of Proline-Based Peptide Models: A Combined Experimental and Molecular Modeling Study.

    Bröhl, Andreas; Albrecht, Benjamin; Zhang, Yong; Maginn, Edward; Giernoth, Ralf


    The influence of three sodium salts, covering a wide range of the Hofmeister series, on the conformation of three proline-based peptide models in aqueous solution is examined using a combination of nuclear magnetic resonance spectroscopy and molecular dynamics simulations. The anions preferentially interact with the cis conformers of the peptide models, which is rationalized by the respective electrostatic potential surfaces. These preferred interactions have a strong impact on the thermodynamics of the cis/trans equilibria, leading to a higher population of the cis conformers. In distinct cases, these equilibria are nearly independent of temperature, showing that the salts are also able to stabilize the conformers over wide temperature ranges.

  9. Catalytic Transformation of Fructose and Sucrose to HMF with Proline-Derived Ionic Liquids under Mild Conditions

    Hu Li


    Full Text Available L-Proline derived ionic liquids (ILs used as both solvent and catalyst were efficient for transformation of fructose and sucrose to 5-hydroxymethylfurfural (HMF in the presence of water. Response surface methodology (RSM was employed to optimize fructose dehydration process, and a maximum HMF yield of 73.6% could be obtained at 90°C after 50 min. The recycling of the IL exhibited an almost constant activity during five successive trials, and a possible reaction mechanism for the dehydration of fructose to HMF was proposed.

  10. A benzotriazole-mediated route to protected marine-derived hetero-2,5-diketopiperazines containing proline.

    Nsengiyumva, Olivier; Hamedzadeh, Sadra; McDaniel, James; Macho, Jocelyn; Simpson, Grant; Panda, Siva S; Ha, Khanh; Lebedyeva, Iryna; Faidallah, Hassan M; Al-Mohammadi, Manal Metgen; Hall, C Dennis; Katritzky, Alan R


    A procedure for the cyclization of dipeptidoyl benzotriazolides containing proline derivatives promoted by triethylamine under MW activation is introduced. The reaction is general for a variety of dipeptidoyl benzotriazolides and represents a very practical and convenient method for the preparation of Pro- or Hyp-derived 2,5-diketopiperazines (2,5-DKPs) and bis-DKPs with a disulfide linker. This method can be used for the construction of 2,5-DKP compound libraries and for the synthesis of natural products with diketopiperazine cores.

  11. Modeling IRA Accumulation and Withdrawals

    Sabelhaus, John


    Empirical analysis of IRA accumulation and withdrawal patterns is limited because information about IRA balances and flows is not available for a sample of taxpayers. This paper combines survey data on IRA balances with individual tax return data on IRA flows to study IRA accumulation and withdrawal patterns across cohorts. The analysis shows that IRA rules such as penalties for early withdrawals and minimum distribution requirements have predictable effects on IRA flows. The estimated propen...

  12. An early nodulin-like protein accumulates in the sieve element plasma membrane of Arabidopsis

    Khan, Junaid A.; Wang, Qi; Sjölund, Richard D.


    that have a similar overall domain structure of an amino-terminal signal peptide, plastocyanin-like copper-binding domain, proline/serine-rich domain, and carboxy-terminal hydrophobic domain. The amino- and carboxy-terminal domains of the 21.5-kD sieve element-specific ENOD are posttranslationally cleaved...... from the precursor protein, resulting in a mature peptide of approximately 15 kD that is attached to the sieve element plasma membrane via a carboxy-terminal glycosylphosphatidylinositol membrane anchor. Many of the Arabidopsis ENOD-like proteins accumulate in gametophytic tissues, whereas in both...... floral and vegetative tissues, the sieve element-specific ENOD is expressed only within the phloem. Members of the ENOD subfamily of the cupredoxin superfamily do not appear to bind copper and have unknown functions. Phenotypic analysis of homozygous T-DNA insertion mutants for the gene At3g20570 shows...

  13. Jasmonic acid/methyl jasmonate accumulate in wounded soybean hypocotyls and modulate wound gene expression.

    Creelman, R A; Tierney, M L; Mullet, J E


    Jasmonic acid (JA) and its methyl ester, methyl jasmonate (MeJA), are plant lipid derivatives that resemble mammalian eicosanoids in structure and biosynthesis. These compounds are proposed to play a role in plant wound and pathogen responses. Here we report the quantitative determination of JA/MeJA in planta by a procedure based on the use of [13C,2H3]MeJA as an internal standard. Wounded soybean (Glycine max [L] Merr. cv. Williams) stems rapidly accumulated MeJA and JA. Addition of MeJA to soybean suspension cultures also increased mRNA levels for three wound-responsive genes (chalcone synthase, vegetative storage protein, and proline-rich cell wall protein) suggesting a role for MeJA/JA in the mediation of several changes in gene expression associated with the plants' response to wounding.

  14. Unscented Kalman filter with parameter identifiability analysis for the estimation of multiple parameters in kinetic models

    Baker Syed; Poskar C; Junker Björn


    Abstract In systems biology, experimentally measured parameters are not always available, necessitating the use of computationally based parameter estimation. In order to rely on estimated parameters, it is critical to first determine which parameters can be estimated for a given model and measurement set. This is done with parameter identifiability analysis. A kinetic model of the sucrose accumulation in the sugar cane culm tissue developed by Rohwer et al. was taken as a test case model. Wh...

  15. Effect of priming on growth, biochemical parameters and mineral composition of different cultivars of coriander (Coriandrum sativum L. under salt stress

    Ben Fredj Meriem


    Full Text Available At Higher Institute of Agriculture of Chott Mariem, Sousse, Tunisia, this study was conducted to evaluate the interactive effect of salinity and seed priming on coriander. The experiment was carried out in completely randomized design with three replications consisting of four coriander genotypes (Tunisian cv, Algerian cv, Syrian cv and Egyptian cv at two seed conditions (seed priming with 4 g/l NaCl for 12h or no seed priming. Results revealed that seed priming and salinity had significantly (p≤0.05 affected all the parameters under study. On the first hand, salinity stress had adversely affected growth, chlorophyll content, mineral composition (K+ and Ca2+ of coriander in all genotypes. Also, it activated Na+ accumulation and synthesis of proline, soluble sugars and proteins. However, seed priming with NaCl had diminished the negative impact of salt stress in all cultivars and primed plants showed better response to salinity compared to unprimed plants. Maximum values were recorded in tolerant cultivar which is Tunisian one whereas minimum values were noted in sensitive cultivar (Algerian cv.

  16. Effect of proline on biochemical and molecular mechanisms in lettuce (Lactuca sativa L.) exposed to UV-B radiation.

    Aksakal, Ozkan; Tabay, Dilruba; Esringu, Aslıhan; Icoglu Aksakal, Feyza; Esim, Nevzat


    The purpose of the present study was to evaluate the role of proline (Pro) in relieving UV-B radiation-induced oxidative stress in lettuce. Lettuce seedlings were exposed to 3.3 W m(-2) UV-B radiation for 12 h after pre-treatment sprayed with 20 mM Pro. The data for malondialdehyde (MDA), hydrogen peroxide (H2O2), endogenous Pro level, the activities of antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD)], total phenolic concentration, antioxidant capacity, expression of phenylalanine ammonia lyase (PAL), γ-tocopherol methyltransferase (γ-TMT) and proline dehydrogenase (ProDH) genes, phytohormone levels such as abscisic acid (ABA), gibberellic acid (GA), indole acetic acid (IAA) and salicylic acid (SA), soluble sugars and organic acids were recorded. It was found that Pro alleviated the oxidative damage in the seedlings of lettuce as demonstrated by lower lipid peroxidation and H2O2 content, increasing the endogenous Pro level, the activity of antioxidant enzymes, total phenolic concentration and the antioxidant capacity. Additionally, it was revealed that exogenous application of Pro enhanced the levels of GA, IAA, the concentrations of soluble sugars and organic acids and expressions of PAL, γ-TMT and ProDH genes as compared to the control. The results obtained in this study suggest that pre-treatment with exogenous Pro provides important contributions to the increase in the UV-B tolerance of lettuce by regulating the biochemical mechanisms of UV-B response.

  17. Proline substitution of dimer interface β-strand residues as a strategy for the design of functional monomeric proteins.

    Joseph, Prem Raj B; Poluri, Krishna Mohan; Gangavarapu, Pavani; Rajagopalan, Lavanya; Raghuwanshi, Sandeep; Richardson, Ricardo M; Garofalo, Roberto P; Rajarathnam, Krishna


    Proteins that exist in monomer-dimer equilibrium can be found in all organisms ranging from bacteria to humans; this facilitates fine-tuning of activities from signaling to catalysis. However, studying the structural basis of monomer function that naturally exists in monomer-dimer equilibrium is challenging, and most studies to date on designing monomers have focused on disrupting packing or electrostatic interactions that stabilize the dimer interface. In this study, we show that disrupting backbone H-bonding interactions by substituting dimer interface β-strand residues with proline (Pro) results in fully folded and functional monomers, by exploiting proline's unique feature, the lack of a backbone amide proton. In interleukin-8, we substituted Pro for each of the three residues that form H-bonds across the dimer interface β-strands. We characterized the structures, dynamics, stability, dimerization state, and activity using NMR, molecular dynamics simulations, fluorescence, and functional assays. Our studies show that a single Pro substitution at the middle of the dimer interface β-strand is sufficient to generate a fully functional monomer. Interestingly, double Pro substitutions, compared to single Pro substitution, resulted in higher stability without compromising native monomer fold or function. We propose that Pro substitution of interface β-strand residues is a viable strategy for generating functional monomers of dimeric, and potentially tetrameric and higher-order oligomeric proteins. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Assessment of the impact of Aluminum on germination, early growth and free proline content in Lactuca sativa L.

    Silva, Patrícia; Matos, Manuela


    Aluminum (Al) toxicity is a major problem in crop production on acid soils. The use of industrial or municipal wastewaters, which may be contaminated with metals, for irrigation in agriculture is common over the world. This action can increase the concentration of these agents in the soil and decrease crops yields. In order to evaluate the toxicological effects of recommended Al levels in irrigation water, under acidic conditions, on lettuce, seeds of two cultivars ("cv Reine de Mai" and "cv White Boston") were exposed to five different Al concentrations (0, 0.05, 0.5, 5 and 20mg/L) and germination percentage, root and shoot lengths were measured. Also, the germination rate and the vigor index were calculated, and the proline content was estimated for all concentrations. Results showed that seed germination was not negatively affected by Al, but the germination rate decreased in both cultivars. For the other factors analyzed, with the exception of 20mg/L concentration for "cv White Boston", Al induced, in general, negative effects including the content of proline that increased in the seeds that were exposed to this metal. The "cv Reine de Mai" was more sensitive for the analyzed concentrations than the other cultivar.. The results indicated that even recommended Al concentrations for irrigation, under acidic conditions, can interfere negatively in seed germination and seedling establishment and possibly with crop production. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. The designer proline-rich antibacterial peptide A3-APO prevents Bacillus anthracis mortality by deactivating bacterial toxins.

    Otvos, Laszlo; Flick-Smith, Helen; Fox, Marc; Ostorhazi, Eszter; Dawson, Raymond M; Wade, John D


    Proline-rich antibacterial peptides protect experimental animals from bacterial challenge even if they are unable to kill the microorganisms in vitro. Their major in vivo modes of action are inhibition of bacterial protein folding and immunostimulation. Here we investigated whether the proline-rich antibacterial peptide dimer A3-APO was able to inhibit Bacillus cereus enterotoxin production in vitro and restrict the proliferation of lethal toxin-induced Bacillus anthracis replication in mouse macrophages. After 24 h incubation, peptide A3-APO and its single chain metabolite reduced the amount of properly folded B. cereus diarrhoeal enterotoxin production in a concentration-dependent manner leading to only 10-25% of the original amount of toxin detectable by a conformation-sensitive immunoassay. Likewise, after 4 h incubation, A3-APO restricted the proliferation of B. anthracis in infected macrophages by 40-45% compared to untreated cells both intracellularly and in the extracellular cell culture milieu. Although the peptide had a minimal inhibitory concentration of >512 mg/L against B. anthracis in vitro, in systemic mouse challenge models it improved survival by 20- 37%, exhibiting statistically significant cumulative efficacy when administered at 3x5 mg/kg intraperitoneally or intramuscularly. We hypothesize that the activity in isolated murine macrophages and in vivo is due to deactivation of bacterial toxins. Bacterial protein folding inhibition in synergy with other types of antimicrobial modes offers a remarkable novel strategy in combating resistant or life-threatening infections.

  20. Effect of proline analogues on activity of human prolyl hydroxylase and the regulation of HIF signal transduction pathway.

    Xiaoyan Ma

    Full Text Available Hypoxia inducible factor 1 (HIF-1 plays a pivotal role in cellular responses to hypoxia. Prolyl hydroxylase 3 (PHD3 degrades HIF-1α under normoxic conditions through the hydroxylation of HIF-1α for proteolysis. Inhibiting PHD3 activity is crucial for up-regulating HIF-1α, thereby acting as a potential target for treating hypoxia-related diseases. In this study, two proline analogues (PA1 and PA2 were screened as PHD3 inhibitors with apparent EC50 values of 1.53 and 3.17 µM respectively, indicating good inhibition potency. Nine proteins, significantly regulated by PA1, were identified using 2-DE coupled with MALDI-TOF/TOF MS. Pyruvate kinase isozymes M1/M2 (PKM and alpha-enolase 1 (ENO1, which are key modulators of glycolysis, are directly regulated by HIF-1α. Moreover, VEGF, a signal protein stimulating angiogenesis, was strongly promoted by PA1. Our findings suggest that PA1 stabilized HIF-1α as well as up-regulated glycolysis and angiogenesis proteins. Herein, for the first time, we systematically studied proline analogue PA1 as a PHD3 inhibitor, which provides innovative evidence for the treatment of HIF-related diseases.

  1. Alterations in antioxidant enzyme activities and proline content in pea leaves under long-term drought stress.

    Karataş, Ilhami; Öztürk, Lokman; Demir, Yavuz; Unlükara, Ali; Kurunç, Ahmet; Düzdemir, Oral


    The effects of long-term drought stress on chlorophyll, proline, protein and hydrogen peroxide (H2O2) contents, malondialdehyde (MDA) in terms of lipid peroxidation and on the changes in the activities of antioxidant enzymes such as superoxide dismutase (SOD, EC, ascorbate peroxidase (APX, EC, catalase (CAT, EC and peroxidase (POX; EC in the leaves of pea (Pisum sativum L.) were studied in field conditions. Chlorophyll and protein contents in leaves decreased significantly with increased drought stress. The proline content increased markedly under water deficit. MDA amounts were elevated as a result of water shortage, whereas H(2)O(2) content changed slightly in pea leaves exposed to drought stress. Drought stress markedly enhanced the activities of SOD, CAT and POX but slightly changed the activity of APX. We conclude that in field conditions, long-term water shortage increased the susceptibility to drought in peas.

  2. Growth and spectroscopy studies of ADP single crystals with L-proline and L-arginine amino acids

    Ben Brahim, F., E-mail: [Laboratoire des Sciences des Materiaux et de l' Environnement (MES-Lab), Faculte des Sciences de Sfax, Route de Soukra km 3.5, BP 1171, 3000 Sfax (Tunisia); Bulou, A. [Laboratoire de Physique de l' Etat Condense (LPEC), UMR CNRS no 6087, Faculte des Sciences et Techniques, Universite du Maine, 72085 Le Mans Cedex 09 (France)


    Highlights: {yields} ADP single crystals were grown with L-proline and L-arginine as guest molecules. {yields} This enhances some properties: piezo-electric, NLO, dielectric and antiferroelectic. {yields} Such compounds can be successfully used for hydrogen bonding studies. - Abstract: Ammonium dihydrogen orthophosphate (ADP) single crystals were grown with L-proline and L-arginine amino acids as additives at 4% molar concentrations. X-ray diffractograms, vibrational FT-IR, and Raman and NMR MAS of {sup 31}P and {sup 1}H spectra of pure and treated compounds were recorded at room temperature and interpreted. A shift of the wavenumbers of some internal modes of both PO{sub 4} and NH{sub 4} tetrahedrons species was observed. At the same time, some characteristic vibrational modes of N-H and C-H bonds of the organic molecules appeared in IR spectra. However, NMR results show a little shift of the resonance of the phosphorus and hydrogen nucleus, associated with some shoulders for this latter. These observations indicate that the host-additives interactions were achieved.

  3. A comparison of the Maillard reactivity of proline to other amino acids using pyrolysis-molecular beam mass spectrometry

    Luc Moens; Robert J. Evans; Michael J. Looker; Mark R. Nimlos [National Bioenergy Center, Golden, CO (United States). National Renewable Energy Laboratory


    Maillard chemistry, or the low temperature condensation of amino acids and carbohydrates, is shown to be relevant to the practical regime of biomass pyrolysis and leads to dramatic changes in low-temperature volatile products and residual solid structure. Mixtures of amino acids and glucose were subjected to a two-temperature heating sequence (5 min each at 170 and 325{sup o}C) and the volatile products analyzed by molecular beam mass spectrometry. Significant volatile yield was observed from the mixtures at 170{sup o}C where neither amino acids nor glucose generated volatile material in the time frame studied. Proline was the most active of the amino acids studied. Volatile products at low temperature included the diketopiperazine, which were generated in higher yields than from proline alone. Also generated were Maillard condensation such as 1-(1{prime}-pyrrolidinyl)-2-propanone. These products were also generated at 325{sup o}C, but in addition, the mass spectra included evidence for the direct formation of nitrogen-containing aromatics. These observations are discussed in relation to known Maillard chemistry. 37 refs., 8 figs., 1 tab.

  4. Effects of dipeptides containing the amino acid, proline on the chemotaxis of Tetrahymena pyriformis. Evolutionary conclusions on the formation of hormone receptors and hormones.

    Köhidai, L; Soós, P; Csaba, G


    Our investigations demonstrate that proline-containing dipeptides can provoke a chemosensory response from the unicellular Tetrahymena pyriformis. The chemotactic effects of the dipeptides have a close relationship with the side chain and the lipophilicity of the amino-terminal amino acid. Comparison of 'mirror' variants of proline-containing dipeptides points to the fact that dipeptides with small side chain and non-polar character amino acids (Gly-Pro, Ala-Pro) are preferred on the amino-terminal end. In the case of amino acids with very variable side chains, small (Pro-Gly) and the large side chain and non-polar character amino acids (Pro-Leu, Pro-Phe) on the carboxyl-terminal end can induce significant chemotactic responses. With valine on any terminus the proline-containing dipeptide induced a weak repellent effect.

  5. Investigation on structural, optical, thermal, mechanical and dielectric properties of L-proline cadmium chloride monohydrate single crystals: An efficient NLO material

    Singh, Preeti; Hasmuddin, Mohd [Crystal Growth and XRD Lab, Department of Physics, Jamia Millia Islamia, New Delhi 110 025 (India); Shakir, Mohd [Crystal Growth and XRD Lab, Department of Physics, Jamia Millia Islamia, New Delhi 110 025 (India); CSIR-National Physical Laboratory, New Delhi 110 012 (India); Department of Physics, ARSD College, University of Delhi, New Delhi 110021 (India); Vijayan, N. [Department of Physics, Kakatiya University, Warangal 506 009 (India); Abdullah, M.M. [Crystal Growth and XRD Lab, Department of Physics, Jamia Millia Islamia, New Delhi 110 025 (India); Advanced Materials and Nano-research Centre (AMNC), Faculty of Science and Arts, Najran University, P.O. Box-1988, Najran 11001 (Saudi Arabia); Ganesh, V. [Department of Physics, Kakatiya University, Warangal 506 009 (India); Wahab, M.A., E-mail: [Crystal Growth and XRD Lab, Department of Physics, Jamia Millia Islamia, New Delhi 110 025 (India)


    In the present work, we have grown single crystals of L-proline cadmium chloride monohydrate (LPCCM) by slow evaporation solution technique (SEST) at room temperature and recorded their live growth kinetics with the help of inverted microscope. Crystal size at various stages of growth and its corresponding morphology was also recorded. Powder X-ray diffraction (PXRD) analysis of LPCCM single crystals confirmed the orthorhombic structure. Respective values of crystallite size, strain and dislocation density have been calculated using PXRD data. Metal complex coordination of the single crystal is studied by FTIR spectroscopic. The optical properties of the grown crystals were investigated through UV–VIS spectroscopic studies and shows that the crystals have very low absorption in entire characterized wavelength range 200–800 nm. The optical band gap was calculated and found to be ∼5.6 eV. Optical constants of the material is determined by theoretical calculations. The chemical etching study was also carried out to study the density of defects in the grown crystals. The photoluminous excitation and emission spectra and thermal property by TGA/DTA curve were recorded. Further, the mechanical properties have been studied using Vicker's microhardness tester as well as many parameters such as fracture toughness (K{sub c}), Brittleness index (B{sub i}) and yield strength (σ{sub ν}) are presented. Dielectric studies have been carried out with varying frequency and temperatures. - Highlights: • The morphology of LPCCM crystal was observed during growth under inverted microscope. • Observation also shows defects also grows with the growth of crystal. • Positive temperature coefficient is observed. • The optical study shows its suitability for photonics and optical applications.

  6. Ternary geochemical-tracing system in natural gas accumulation


    The establishment of geochemical-tracing system of gas generation and accumulation is helpful to re-elucidating the gas migration and accumulation in time and space. To deduce the complex process of gas accumulation, a ternary geochemical-tracing system is set up, according to stable isotope inheritance of source rocks, kinetic fractionation of stable isotopes, time-accumulating effect of noble gas isotopes, mantle-derived volatile inheritance, and organic molecule inheritance of light hydrocarbons and thermally kinetic fractionation in their generation, in combination with the previous achievements of gas geochemistry and geochemical parameters of gas-source correlation. There are tight interactions for the geochemical parameters with much information about parent inheritance and special biomarkers, in which they are confirmed each other, reciprocally associated and preferentially used for the requirement so that we can use these geochemical parameters to effectively demonstrate the sources of natural gas, sedimentary environments and thermal evolution of source rocks, migration and accumulation of natural gas, and rearrangement of natural gas reservoirs. It is necessary for the ternary geochemical-tracing system to predict the formation of high efficient gas reservoir and their distribution in time and space.

  7. Allosteric reversion of Haemophilus influenzae β-carbonic anhydrase via a proline shift.

    Hoffmann, Katherine M; Million-Perez, H Rachael; Merkhofer, Richard; Nicholson, Hilary; Rowlett, Roger S


    Haemophilus influenzae β-carbonic anhydrase (HICA) has been reverse-engineered in the allosteric site region to resemble the nonallosteric Pisum sativum enzyme in order to identify critical features of allostery and intersusbunit communication. Three variants (W39V/G41A, P48S/A49P, and W39V/G41A/P48S/A49P) were identified, through a comparison with a crystal structure of nonallosteric P. sativum β-carbonic anhydrase (PSCA, PDB 1EKJ ), to potentially revert HICA to a nonallosteric enzyme. The W39V/G41A and P48S/A49P mutations decreased the apparent kcat/Km proton dependence from 4 to 2 and 1, respectively, increasing the overall maximal kcat/Km to 16 ± 2 μM(-1) s(-1) (380% of wild type) and 17 ± 3 μM(-1) s(-1) (405% of wild type). The pKa values of the metal-bound water molecule based on the pH-rate profile kinetics (8.32 ± 0.04 for W39V/G41A and 8.3 ± 0.1 for P48S/A49P) were also slightly higher than that for the wild-type enzyme (7.74 ± 0.04). The P48S/A49P variant has lost all pH-rate cooperativity. The W39V/G41A/P48S/A49P variant's kinetics were unusual and were fit with a log-linear function with a slope 0.9 ± 0.2. The crystal structure of the W39V/G41A variant revealed an active site very similar to the T-state wild-type oligomer with bicarbonate trapped in the escort site. By contrast, the X-ray crystal structure of a proline shift variant (P48S/A49P) reveals that it has adopted an active site conformation nearly identical to that of nonallosteric β-carbonic anhydrase (R-state) for one chain, including a tight association with the dimer-exchanged N-terminal helices; the second chain in the asymmetric unit is associated in a biologically relevant oligomer, but it adopts a T-state conformation that is not capped by dimer-exchanged N-terminal helices. The hybrid R/T nature of HICA P48S/A49P structurally recapitulates the interruption of pH-rate cooperativity observed for this variant. Comparison of the conformations of the R and T chains of P48S/A49P

  8. Enhancement of the proline and nitric oxide synthetic pathway improves fermentation ability under multiple baking-associated stress conditions in industrial baker's yeast

    Sasano Yu


    Full Text Available Abstract Background During the bread-making process, industrial baker's yeast, mostly Saccharomyces cerevisiae, is exposed to baking-associated stresses, such as air-drying and freeze-thaw stress. These baking-associated stresses exert severe injury to yeast cells, mainly due to the generation of reactive oxygen species (ROS, leading to cell death and reduced fermentation ability. Thus, there is a great need for a baker's yeast strain with higher tolerance to baking-associated stresses. Recently, we revealed a novel antioxidative mechanism in a laboratory yeast strain that is involved in stress-induced nitric oxide (NO synthesis from proline via proline oxidase Put1 and N-acetyltransferase Mpr1. We also found that expression of the proline-feedback inhibition-less sensitive mutant γ-glutamyl kinase (Pro1-I150T and the thermostable mutant Mpr1-F65L resulted in an enhanced fermentation ability of baker's yeast in bread dough after freeze-thaw stress and air-drying stress, respectively. However, baker's yeast strains with high fermentation ability under multiple baking-associated stresses have not yet been developed. Results We constructed a self-cloned diploid baker's yeast strain with enhanced proline and NO synthesis by expressing Pro1-I150T and Mpr1-F65L in the presence of functional Put1. The engineered strain increased the intracellular NO level in response to air-drying stress, and the strain was tolerant not only to oxidative stress but also to both air-drying and freeze-thaw stresses probably due to the reduced intracellular ROS level. We also showed that the resultant strain retained higher leavening activity in bread dough after air-drying and freeze-thaw stress than that of the wild-type strain. On the other hand, enhanced stress tolerance and fermentation ability did not occur in the put1-deficient strain. This result suggests that NO is synthesized in baker's yeast from proline in response to oxidative stresses that induce ROS

  9. Effects of cadmium stress on growth, metal accumulation and ...



    Mar 15, 2012 ... production of MDA, damage of cell membrane, accumu- lation of proline, soluble ..... of undesirable cyanobacteria (Shen et al., 2008). In addition, enzymatic .... Structure and function of metal chelators produced by plants. Cell.

  10. Supergranular Parameters

    Udayashankar, Paniveni


    I study the complexity of supergranular cells using intensity patterns from Kodaikanal solar observatory. The chaotic and turbulent aspect of the solar supergranulation can be studied by examining the interrelationships amongst the parameters characterizing supergranular cells namely size, horizontal flow field, lifetime and physical dimensions of the cells and the fractal dimension deduced from the size data. The findings are supportive of Kolmogorov's theory of turbulence. The Data consists of visually identified supergranular cells, from which a fractal dimension 'D' for supergranulation is obtained according to the relation P α AD/2 where 'A' is the area and 'P' is the perimeter of the supergranular cells. I find a fractal dimension close to about 1.3 which is consistent with that for isobars and suggests a possible turbulent origin. The cell circularity shows a dependence on the perimeter with a peak around (1.1-1.2) x 105 m. The findings are supportive of Kolmogorov's theory of turbulence.

  11. Completeness of the Accumulation Calculus

    虞慧群; 孙永强; 等


    The accumulation calculs(AC for short)is an interval based temporal logic to specify and reason about hybrid real-time systems.This paper presents a formal proof system for AC,and proves that the system is complete relative to that of Interval Temporal Logic(ITL for short)on real domain.

  12. Detection of discriminative sequence patterns in the neighborhood of proline cis peptide bonds and their functional annotation

    Papaloukas Costas


    Full Text Available Abstract Background Polypeptides are composed of amino acids covalently bonded via a peptide bond. The majority of peptide bonds in proteins is found to occur in the trans conformation. In spite of their infrequent occurrence, cis peptide bonds play a key role in the protein structure and function, as well as in many significant biological processes. Results We perform a systematic analysis of regions in protein sequences that contain a proline cis peptide bond in order to discover non-random associations between the primary sequence and the nature of proline cis/trans isomerization. For this purpose an efficient pattern discovery algorithm is employed which discovers regular expression-type patterns that are overrepresented (i.e. appear frequently repeated in a set of sequences. Four types of pattern discovery are performed: i exact pattern discovery, ii pattern discovery using a chemical equivalency set, iii pattern discovery using a structural equivalency set and iv pattern discovery using certain amino acids' physicochemical properties. The extracted patterns are carefully validated using a specially implemented scoring function and a significance measure (i.e. log-probability estimate indicative of their specificity. The score threshold for the first three types of pattern discovery is 0.90 while for the last type of pattern discovery 0.80. Regarding the significance measure, all patterns yielded values in the range [-9, -31] which ensure that the derived patterns are highly unlikely to have emerged by chance. Among the highest scoring patterns, most of them are consistent with previous investigations concerning the neighborhood of cis proline peptide bonds, and many new ones are identified. Finally, the extracted patterns are systematically compared against the PROSITE database, in order to gain insight into the functional implications of cis prolyl bonds. Conclusion Cis patterns with matches in the PROSITE database fell mostly into two

  13. Accumulation of heavy metals using Sorghum sp.

    Soudek, Petr; Petrová, Šarka; Vaňková, Radomíra; Song, Jing; Vaněk, Tomaš


    The essential requirement for the effective phytoremediation is selection of a plant species which should be metal tolerant, with high biomass production and known agronomic techniques. The above mentioned criteria are met by crop plant sorghum (Sorghum bicolor). The response of hydroponically grown S. bicolor plants to cadmium and zinc stress was followed. The impact of metal application on physiological parameters, including changes in chlorophylls contents and antioxidative enzymes activities, was followed during the stress progression. Cadmium and zinc were accumulated primarily in the roots of sorghum plants. However, elevation of metal concentrations in the media promoted their transfer to the shoots. Toxic effects of metals applied at lower concentrations were less serious in the shoots in comparison with their influence to the roots. When applied at higher concentrations, transfer of the metals into the leaves increased, causing growth reduction and leading to Chl loss and metal-induced chlorosis. Moreover, higher metal levels in the roots overcame the quenching capacity of peroxidase and glutathione transferase, which was associated with reduction of their activities. Fortification of antioxidant system by addition of glutathione significantly increased the accumulation of cadmium in the roots as well as in the shoots at the highest cadmium concentration applied.

  14. Association of the leucine-7 to proline-7 variation in the signal sequence of neuropeptide Y with major depression

    Koefoed, Pernille; Woldbye, David Paul Drucker; Hansen, Thomas v.O;


    Objective: There is clear evidence of a genetic component in major depression, and several studies indicate that neuropeptide Y (NPY) could play an important role in the pathophysiology of the disease. A well-known polymorphism encoding the substitution of leucine to proline in the signal peptide...... sequence of NPY (Leu7Pro variation) was previously found to protect against depression. Our study aimed at replicating this association in a large Danish population with major depression. Method: Leu7Pro was studied in a sample of depressed patients and ethnically matched controls, as well as psychiatric.......46) as compared to ethnically matched controls (3.8%, n = 2912), while schizophrenia patients (4.1%, n = 503) did not differ. In vitro, the Pro7 substitution appeared to be associated with reduced levels of NPY without affecting its mRNA level. Conclusion: The Leu7Pro variation may increase the risk of major...

  15. Practical synthesis of enantiomerically pure beta2-amino acids via proline-catalyzed diastereoselective aminomethylation of aldehydes.

    Chi, Yonggui; English, Emily P; Pomerantz, William C; Horne, W Seth; Joyce, Leo A; Alexander, Lane R; Fleming, William S; Hopkins, Elizabeth A; Gellman, Samuel H


    Proline-catalyzed diastereoselective aminomethylation of aldehydes using a chiral iminium ion, generated from a readily prepared precursor, provides alpha-substituted-beta-amino aldehydes with 85:15 to 90:10 dr. The alpha-substituted-beta-amino aldehydes can be reduced to beta-substituted-gamma-amino alcohols, the major diastereomer of which can be isolated via crystallization or column chromatography. The amino alcohols are efficiently transformed to protected beta2-amino acids, which are valuable building blocks for beta-peptides, natural products, and other interesting molecules. Because conditions for the aminomethylation and subsequent reactions are mild, beta2-amino acid derivatives with protected functional groups in the side chain, such as beta2-homoglutamic acid, beta2-homotyrosine, and beta2-homolysine, can be prepared in this way. The synthetic route is short, and purifications are simple; therefore, this method enables the preparation of protected beta2-amino acids in useful quantities.

  16. Synthesis, crystal growth and characterizations of bis ( l-proline) cadmium iodide: a new semi-organic nonlinear optical material

    Boopathi, K.; Jagan, R.; Ramasamy, P.


    Novel semi-organic single crystals of bis ( l-proline) cadmium iodide (BLPC) were grown by slow evaporation technique. The crystal structure was determined by single-crystal X-ray diffraction studies. Single-crystal X-ray diffraction study shows that [BLPC] crystallizes in orthorhombic system with space group P212121. 1H NMR and 13C NMR studies were conducted for the grown crystal. Functional groups present in the compound were identified by FTIR spectral studies. The UV-Vis-NIR spectrum was studied to analyse the optical properties of the grown crystals. Thermogravimetric analysis was carried out to study thermal behaviour of the materials. Vickers microhardness measurement was carried out for different loads. Etching studies were carried out using water as etchant. The second harmonic generation efficiency was determined by the Kurtz powder method and it was found to be higher than that of potassium dihydrogen phosphate.

  17. Association of the leucine-7 to proline-7 variation in the signal sequence of neuropeptide Y with major depression

    Koefoed, Pernille; Woldbye, David P. D.; Hansen, Thomas v. O.


    Objective: There is clear evidence of a genetic component in major depression, and several studies indicate that neuropeptide Y (NPY) could play an important role in the pathophysiology of the disease. A well-known polymorphism encoding the substitution of leucine to proline in the signal peptide...... sequence of NPY (Leu7Pro variation) was previously found to protect against depression. Our study aimed at replicating this association in a large Danish population with major depression. Method: Leu7Pro was studied in a sample of depressed patients and ethnically matched controls, as well as psychiatric...... disease controls with schizophrenia. Possible functional consequences of Leu7Pro were explored in vitro. Results: In contrast to previous studies, Pro7 appeared to be a risk allele for depression, being significantly more frequent in the depression sample (5.5 n = 593; p = 0.009; odds ratio, OR: 1...

  18. Discovery, cloning and characterisation of proline specific prolyl endopeptidase, a gluten degrading thermo-stable enzyme from Sphaerobacter thermophiles

    Shetty, Radhakrishna; Vestergaard, Mike; Jessen, Flemming


    Gluten free products have emerged during the last decades, as a result of a growing public concern and technological advancements allowing gluten reduction in food products. One approach is to use gluten degrading enzymes, typically at low or ambient temperatures, whereas many food production...... processes occur at elevated temperature. We present in this paper, the discovery, cloning and characterisation of a novel recombinant thermostable gluten degrading enzyme, a proline specific prolyl endoprotease (PEP) from Sphaerobacter thermophiles. The molecular mass of the prolyl endopeptidase...... at 63 °C was higher than 75 %. The enzyme was activated and stabilized by Co2+ and inhibited by Mg2+, K+ and Ca2+ followed by Zn2+, Na+, Mn2+, Al3+, and Cu2+. The Km and kcat values of the purified enzyme for different substrates were evaluated. The ability to degrade immunogenic gluten peptides...

  19. Kinetic Mechanism of Uracil Phosphoribosyltransferase from Escherichia coli and Catalytic Importance of the Conserved Proline in the PRPP Binding Site

    Lundegaard, Claus; Jensen, Kaj Frank


    catalytic properties with the properties of the wild-type protein. We found that UPRTase of E. coli obeyed the kinetics of a sequential mechanism with the binding of PRPP preceding the binding of uracil. The basic kinetic constants were derived from initial velocity measurements, product inhibition......, and ligand binding assays. The change of Pro 131 to Asp caused a 50-60-fold reduction of the catalytic rate (kcat) in both directions of the reaction and approximately a 100-fold increase in the KM for uracil. The KM for PRPP was strongly diminished by the mutation, but kcat/KM,PRPP and the dissociation...... constant (KD,PRPP) were nearly unaffected. We conclude that the proline in the PRPP binding site of UPRTase is of only little importance for binding of PRPP to the free enzyme, but is critical for binding of uracil to the enzyme-PRPP complex and for the catalytic rate....

  20. N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M.

    Manconi, Barbara; Cabras, Tiziana; Sanna, Monica; Piras, Valentina; Liori, Barbara; Pisano, Elisabetta; Iavarone, Federica; Vincenzoni, Federica; Cordaro, Massimo; Faa, Gavino; Castagnola, Massimo; Messana, Irene


    In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213.

  1. Salicylic Acid Protects Nitrate Reductase Activity, Growth and Proline in Amaranth and Tomato Plants during Water Deficit

    C. E. Umebese


    Full Text Available Problem statement: Seedlings of Amaranthus hybridus cv. NHAC-3 (large green, amaranth and Lycopersicum esculentum cv. Roma (tomato were subjected to 7 days water stress at Early Vegetative (EV, Late Vegetative (LV, Early Flowering (EF and Late Flowering (LF stages of growth to study the impact on leaf water potential (ψw, Nitrate Reductase Activity (NRA, growth (plant height, shoot and root biomass and proline content of both plants. Approach: Two concentrations of salicylic acid (1 and 3 mM SA were applied to stressed plants to study the level of protection given by SA to the plants. Leaf ψw was significantly reduced (p = 0.05 during stress treatment at nearly all stages of growth in both plants. Leaf ψw was in the range -0.25 to -1.42 (unstressed and -1.45 to -2.02 (stressed in tomato plants while in amaranth it was -0.7 to -1.62 (unstressed and -1.62 to -2.68 (stressed. As 3 mM SA increased leaf ψw to values close to the control (unstressed plants. NRA was significantly (p = 0.05 reduced by stress treatment at the LV stage of amaranth, EF stage of tomato and LF stage of both plants. Results: Thus, the reduction of NRA was more pronounced at the reproductive stage of both plants. As 3 mM SA was effective in maintaining NRA at levels similar to the control in both plants. Stress treatment reduced plant height significantly (p = 0.05 at the vegetative stages of both plants and 3 mM was also effective in keeping plant height similar to the control. Though shoot biomass was affected by water stress, SA treatment was not very effective in preserving the biomass during stress. Root biomass of plants was reduced by stress treatment at the reproductive stage and only tomato plants responded positively to 3 mM SA. Proline content was only slightly increased at all stages of growth in stressed plants but 3 mM SA induced a two-fold increase in proline content at the vegetative stage of tomato (EV and LV and significant increases (p = 0.05 at almost

  2. Improving co-amorphous drug formulations by the addition of the highly water soluble amino acid proline

    Jensen, Katrine Birgitte Tarp; Löbmann, Korbinian; Rades, Thomas;


    by combining the model drug, naproxen (NAP), with an amino acid to physically stabilize the co-amorphous system (tryptophan, TRP, or arginine, ARG) and a second highly soluble amino acid (proline, PRO) for an additional improvement of the dissolution rate. Co-amorphous drug-amino acid blends were prepared......Co-amorphous drug amino acid mixtures were previously shown to be a promising approach to create physically stable amorphous systems with the improved dissolution properties of poorly water-soluble drugs. The aim of this work was to expand the co-amorphous drug amino acid mixture approach...... by ball milling and investigated for solid state characteristics, stability and the dissolution rate enhancement of NAP. All co-amorphous mixtures were stable at room temperature and 40 °C for a minimum of 84 days. PRO acted as a stabilizer for the co-amorphous system, including NAP–TRP, through enhancing...

  3. Application of bis-(L zinc prolinate (II on the synthesis reactions of some 1,4-dyhidropyridines

    Cristiane R. Winck


    Full Text Available The 1,4-Dyhidropyridine (DHP is a class of substances that presents a wide spectrum of biological activities. Among these applications, it can be applied on the treatment of cardiovascular diseases e.g. hypertension. This project proposes the synthesis of some DHPs through multicomponent reactions (one pot using bis-(L zinc prolinate (II as catalyst. Initially the synthesis of the precursor enamine was held using the catalyst and later, there was an addition of cinnmaldehyde to the same reaction media. Such procedure resulted on the formation of the compound of interest (Figure 1. Another important factor is that the ultrasound was used to reach the results, which makes such reactions highly convergent and coherent with the principles of the Green Chemistry.

  4. Essential role of tissue-specific proline synthesis and catabolism in growth and redox balance at low water potential.

    Sharma, Sandeep; Villamor, Joji Grace; Verslues, Paul E


    To better define the still unclear role of proline (Pro) metabolism in drought resistance, we analyzed Arabidopsis (Arabidopsis thaliana) Δ(1)-pyrroline-5-carboxylate synthetase1 (p5cs1) mutants deficient in stress-induced Pro synthesis as well as proline dehydrogenase (pdh1) mutants blocked in Pro catabolism and found that both Pro synthesis and catabolism were required for optimal growth at low water potential (ψ(w)). The abscisic acid (ABA)-deficient mutant aba2-1 had similar reduction in root elongation as p5cs1 and p5cs1/aba2-1 double mutants. However, the reduced growth of aba2-1 but not p5cs1/aba2-1 could be complemented by exogenous ABA, indicating that Pro metabolism was required for ABA-mediated growth protection at low ψ(w). PDH1 maintained high expression in the root apex and shoot meristem at low ψ(w) rather than being repressed, as in the bulk of the shoot tissue. This, plus a reduced oxygen consumption and buildup of Pro in the root apex of pdh1-2, indicated that active Pro catabolism was needed to sustain growth at low ψ(w). Conversely, P5CS1 expression was most highly induced in shoot tissue. Both p5cs1-4 and pdh1-2 had a more reduced NADP/NADPH ratio than the wild type at low ψ(w). These results indicate a new model of Pro metabolism at low ψ(w) whereby Pro synthesis in the photosynthetic tissue regenerates NADP while Pro catabolism in meristematic and expanding cells is needed to sustain growth. Tissue-specific differences in Pro metabolism and function in maintaining a favorable NADP/NADPH ratio are relevant to understanding metabolic adaptations to drought and efforts to enhance drought resistance.

  5. Two novel MYH7 proline substitutions cause Laing Distal Myopathy-like phenotypes with variable expressivity and neck extensor contracture.

    Feinstein-Linial, Miora; Buvoli, Massimo; Buvoli, Ada; Sadeh, Menachem; Dabby, Ron; Straussberg, Rachel; Shelef, Ilan; Dayan, Daniel; Leinwand, Leslie Anne; Birk, Ohad S


    Human skeletal muscles express three major myosin heavy chain (MyHC) isoforms: MyHCIIx (MYH1) in fast type 2B muscle fibers, MyHCIIa (MYH2) in fast type 2A fibers and MyHCI/β-cardiac MyHC (MYH7) in slow type I skeletal fibers and cardiac ventricles. In line with its expression pattern, MYH7 mutations have been reported in association with hypertrophic or dilated cardiomyopathy, skeletal myopathies or a combination of both. We analyzed the clinical and molecular phenotype of two unrelated families of Jewish Moroccan ancestry that presented with apparently autosomal dominant inheritance of progressive Laing-like distal myopathy with non-specific myopathic changes, but uncommon marked contractures and wasting of the neck extensors. Clinical phenotyping, whole exome sequencing and restriction analysis, generation of mutants followed by cell culture transfection and imaging. Using whole exome sequencing we identified in both families two novel heterozygous proline substitutions located in exon 31 of MYH7 within its rod domain: c.4309G>C (p.Ala1437Pro) and c.4301G>C (p.Arg1434Pro). Here we show that the phenotype caused by these mutations includes marked cervical muscle contracture, and report that the severity of the phenotype varies significantly, to the extent of non-penetrance in one of the families. Finally, we provide evidence that both proline substitutions impair myosin self-assembly in non-muscle cells transfected with β-myosin constructs carrying the mutations, but do not prevent incorporation of the mutant molecules into the sarcomere. This study expands our clinical and molecular knowledge of MYH7 rod mutations causing skeletal myopathies, and underscores the importance of discussing disease penetrance during genetic counseling.

  6. Conformation of a group 2 late embryogenesis abundant protein from soybean. Evidence of poly (L-proline)-type II structure.

    Soulages, Jose L; Kim, Kangmin; Arrese, Estela L; Walters, Christina; Cushman, John C


    Late embryogenesis abundant (LEA) proteins are members of a large group of hydrophilic, glycine-rich proteins found in plants, algae, fungi, and bacteria known collectively as hydrophilins that are preferentially expressed in response to dehydration or hyperosmotic stress. Group 2 LEA (dehydrins or responsive to abscisic acid) proteins are postulated to stabilize macromolecules against damage by freezing, dehydration, ionic, or osmotic stress. However, the structural and physicochemical properties of group 2 LEA proteins that account for such functions remain unknown. We have analyzed the structural properties of a recombinant form of a soybean (Glycine max) group 2 LEA (rGmDHN1). Differential scanning calorimetry of purified rGmDHN1 demonstrated that the protein does not display a cooperative unfolding transition upon heating. Ultraviolet absorption and circular dichroism spectroscopy revealed that the protein is in a largely hydrated and unstructured conformation in solution. However, ultraviolet absorption and circular dichroism measurements collected at different temperatures showed that the protein exists in equilibrium between two extended conformational states: unordered and left-handed extended helical or poly (L-proline)-type II structures. It is estimated that 27% of the residues of rGmDHN1 adopt or poly (L-proline)-type II-like helical conformation at 12 degrees C. The content of extended helix gradually decreases to 15% as the temperature is increased to 80 degrees C. Studies of the conformation of the protein in solution in the presence of liposomes, trifluoroethanol, and sodium dodecyl sulfate indicated that rGmDHN1 has a very low intrinsic ability to adopt alpha-helical structure and to interact with phospholipid bilayers through amphipathic alpha-helices. The ability of the protein to remain in a highly extended conformation at low temperatures could constitute the basis of the functional role of GmDHN1 in the prevention of freezing, desiccation

  7. A Novel Topology of Proline-rich Transmembrane Protein 2 (PRRT2): HINTS FOR AN INTRACELLULAR FUNCTION AT THE SYNAPSE.

    Rossi, Pia; Sterlini, Bruno; Castroflorio, Enrico; Marte, Antonella; Onofri, Franco; Valtorta, Flavia; Maragliano, Luca; Corradi, Anna; Benfenati, Fabio


    Proline-rich transmembrane protein 2 (PRRT2) has been identified as the single causative gene for a group of paroxysmal syndromes of infancy, including epilepsy, paroxysmal movement disorders, and migraine. On the basis of topology predictions, PRRT2 has been assigned to the recently characterized family of Dispanins, whose members share the two-transmembrane domain topology with a large N terminus and short C terminus oriented toward the outside of the cell. Because PRRT2 plays a role at the synapse, it is important to confirm the exact orientation of its N and C termini with respect to the plasma membrane to get clues regarding its possible function. Using a combination of different experimental approaches, including live immunolabeling, immunogold electron microscopy, surface biotinylation and computational modeling, we demonstrate a novel topology for this protein. PRRT2 is a type II transmembrane protein in which only the second hydrophobic segment spans the plasma membrane, whereas the first one is associated with the internal surface of the membrane and forms a helix-loop-helix structure without crossing it. Most importantly, the large proline-rich N-terminal domain is not exposed to the extracellular space but is localized intracellularly, and only the short C terminus is extracellular (N cyt/C exo topology). Accordingly, we show that PRRT2 interacts with the Src homology 3 domain-bearing protein Intersectin 1, an intracellular protein involved in synaptic vesicle cycling. These findings will contribute to the clarification of the role of PRRT2 at the synapse and the understanding of pathogenic mechanisms on the basis of PRRT2-related neurological disorders.

  8. Beta-keratins of turtle shell are glycine-proline-tyrosine rich proteins similar to those of crocodilians and birds.

    Dalla Valle, Luisa; Nardi, Alessia; Toni, Mattia; Emera, Deena; Alibardi, Lorenzo


    This study presents, for the first time, sequences of five beta-keratin cDNAs from turtle epidermis obtained by means of 5'- and 3'-rapid amplification of cDNA ends (RACE) analyses. The deduced amino acid sequences correspond to distinct glycine-proline-serine-tyrosine rich proteins containing 122-174 amino acids. In situ hybridization shows that beta-keratin mRNAs are expressed in cells of the differentiating beta-layers of the shell scutes. Southern blotting analysis reveals that turtle beta-keratins belong to a well-conserved multigene family. This result was confirmed by the amplification and sequencing of 13 genomic fragments corresponding to beta-keratin genes. Like snake, crocodile and avian beta-keratin genes, turtle beta-keratins contain an intron that interrupts the 5'-untranslated region. The length of the intron is variable, ranging from 0.35 to 1.00 kb. One of the sequences obtained from genomic amplifications corresponds to one of the five sequences obtained from cDNA cloning; thus, sequences of a total of 17 turtle beta-keratins were determined in the present study. The predicted molecular weight of the 17 different deduced proteins range from 11.9 to 17.0 kDa with a predicted isoelectric point of 6.8-8.4; therefore, they are neutral to basic proteins. A central region rich in proline and with beta-strand conformation shows high conservation with other reptilian and avian beta-keratins, and it is likely involved in their polymerization. Glycine repeat regions, often containing tyrosine, are localized toward the C-terminus. Phylogenetic analysis shows that turtle beta-keratins are more similar to crocodilian and avian beta-keratins than to those of lizards and snakes.

  9. Use of cis-[18F]fluoro-proline for assessment of exercise-related collagen synthesis in musculoskeletal connective tissue.

    Dorthe Skovgaard

    Full Text Available Protein turnover in collagen rich tissue is influenced by exercise, but can only with difficulty be studied in vivo due to use of invasive procedure. The present study was done to investigate the possibility of applying the PET-tracer, cis-[(18F]fluoro-proline (cis-Fpro, for non-invasive assessment of collagen synthesis in rat musculoskeletal tissues at rest and following short-term (3 days treadmill running. Musculoskeletal collagen synthesis was studied in rats at rest and 24 h post-exercise. At each session, rats were PET scanned at two time points following injection of cis-FPro: (60 and 240 min p.i. SUV were calculated for Achilles tendon, calf muscle and tibial bone. The PET-derived results were compared to mRNA expression of collagen type I and III. Tibial bone had the highest SUV that increased significantly (p<0.001 from the early (60 min to the late (240 min PET scan, while SUV in tendon and muscle decreased (p<0.001. Exercise had no influence on SUV, which was contradicted by an increased gene expression of collagen type I and III in muscle and tendon. The clearly, visible uptake of cis-Fpro in the collagen-rich musculoskeletal tissues is promising for multi-tissue studies in vivo. The tissue-specific differences with the highest basal uptake in bone are in accordance with earlier studies relying on tissue incorporation of isotopic-labelled proline. A possible explanation of the failure to demonstrate enhanced collagen synthesis following exercise, despite augmented collagen type I and III transcription, is that SUV calculations are not sensitive enough to detect minor changes in collagen synthesis. Further studies including kinetic compartment modeling must be performed to establish whether cis-Fpro can be used for non-invasive in-vivo assessment of exercise-induced changes in musculoskeletal collagen synthesis.

  10. Cadmium stress in wheat seedlings: growth, cadmium accumulation and photosynthesis

    Ci, Dunwei; Jiang, Dong; Wollenweber, Bernd


    parameters were generally depressed by Cd stress, especially under the high Cd concentrations. Cd concentration and accumulation in both shoots and roots increased with increasing external Cd concentrations. Relationships between corrected parameters of growth, photosynthesis and fluorescence and corrected...... Cd concentrations in shoots and roots could be explained by the regression model Y = K/(1 + exp(a + bX)). Jing 411 was found to be Cd tolerant considering parameters of chlorophyll content, photosynthesis and chlorophyll fluorescence in which less Cd translocation was from roots into shoots. The high...

  11. L-Proline: an efficient N,O-bidentate ligand for copper-catalyzed aerobic oxidation of primary and secondary benzylic alcohols at room temperature.

    Zhang, Guofu; Han, Xingwang; Luan, Yuxin; Wang, Yong; Wen, Xin; Ding, Chengrong


    A novel and highly practical copper-catalyzed aerobic alcohol oxidation system with L-proline as the ligand at room temperature has been developed. A wide range of primary and secondary benzylic alcohols tested have been smoothly transformed into corresponding aldehydes and ketones with high yields and selectivities.

  12. Cul-catalyzed Coupling Reactions of Aryl Iodides with Amides Using L-Proline and KF/Al2O3



    An efficient experimentally simple and inexpensive catalyst system for the selective amidation of aryl iodides using 15 mol% of Cul as catalyst,15 mol% of L-proline as ligand and KF/Al2O3 as a base in toluene is described.

  13. The proline-histidine-rich CDK2/CDK4 interaction region of C/EBPalpha is dispensable for C/EBPalpha-mediated growth regulation in vivo

    Porse, Bo Torben; Pedersen, Thomas Askov; Hasemann, Marie Sigurd


    that a short, centrally located, 15-amino-acid proline-histidine-rich region (PHR) of C/EBPalpha is responsible for the growth-inhibitory function of the protein through its ability to interact with CDK2 and CDK4, thereby inhibiting their activities. Homozygous Cebpa(DeltaPHR/DeltaPHR) (DeltaPHR) mice...

  14. The location of redox-sensitive groups in the carrier protein of proline at the outer and inner surface of the membrane in Escherichia coli

    Poolman, Bert; Konings, Wil N.; Robillard, George T.


    Evidence is presented in this report for the presence of two sets of dithiols associated with proline transport activity in Escherichia coli. One set is located at the outer surface, the other at the inner surface of the cytoplasmic membrane. Treatment of right-side-out membrane vesicles from E. col

  15. Post-Translational Modification and Secretion of Azelaic Acid Induced 1 (AZI1, a Hybrid Proline-Rich Protein from Arabidopsis

    Andrea Pitzschke


    Full Text Available Arabidopsis EARLI-type hybrid proline-rich proteins (HyPRPs consist of a putative N-terminal secretion signal, a proline-rich domain (PRD, and a characteristic eight-cysteine-motif (8-CM. They have been implicated in biotic and abiotic stress responses. AZI1 is required for systemic acquired resistance and it has recently been identified as a target of the stress-induced mitogen-activated protein kinase MPK3. AZI1 gel migration properties strongly indicate AZI1 to undergo major post-translational modifications. These occur in a stress-independent manner and are unrelated to phosphorylation by MAPKs. As revealed by transient expression of AZI1 in Nicotiana benthamiana and Tropaeolum majus, the Arabidopsis protein is similarly modified in heterologous plant species. Proline-rich regions, resembling arabinogalactan proteins point to a possible proline hydroxylation and subsequent O-glycosylation of AZI1. Consistently, inhibition of prolyl hydroxylase reduces its apparent protein size. AZI1 secretion was examined using Arabidopsis protoplasts and seedling exudates. Employing Agrobacterium-mediated leaf infiltration of N. benthamiana, we attempted to assess long-distance movement of AZI1. In summary, the data point to AZI1 being a partially secreted protein and a likely new member of the group of hydroxyproline-rich glycoproteins. Its dual location suggests AZI1 to exert both intra- and extracellular functions.

  16. Soluble Proteins, Proline, Carbohydrates and Na+/K+ Changes in Two Tomato (Lycopersicon esculentum Mill. Cultivars under in vitro Salt Stress

    Fariba Amini


    Full Text Available The effect of NaCl stress on soluble proteins, proline, carbohydrates and Na+/K+ of two tomato (Lycopersicon esculentum Mill. cultivars Isfahani and Shirazy were studied. Seeds were germinated on medium containing only water agar, then transferred to MS medium supplemented with different concentrations of NaCl (0, 40, 80, 120 and 160 mM for 21 days. Increasing of salinity resulted in increasing of soluble proteins in stem & leaf of cv. Isfahani but decreasing in cv. Shirazy. Soluble proteins in roots of both cultivars showed some variations. When concentration of NaCl in the medium was increased proline contents of stem-leaf and roots in both cultivars increased significantly. However, cv. Shirazy showed higher amount of proline level. Proline content in stem-leaf in comparing with roots was higher in two cultivars. In response to increasing of salt concentration of the medium, the average amount of total carbohydrate in stem-leaf of cv. Shirazy increased but, in cv. Isfahani level of carbohydrate decreased. When explants form both cultivars were exposed to the higher concentration of salt the level of carbohydrate in roots increased. High-level salinity decreased the reduced sugars in both cv. either in stem-leaf or roots. Salt stress increased Na+ and decreased K+ content in both cultivars significantly.

  17. Effects of fixation and demineralization on the intensity of autoradiographic labelling over the periodontal ligament of the mouse incisor after administration of [3H]-proline

    Beertsen, W.; Tonino, G.J.M.


    The effect of different histological procedures on the autoradiographic grain count over the periodontal ligament was studied quantitatively in autoradiographs made eight hours after administration of [3H]-proline. The lower jaws of 9 mice were fixed in Bouin's fixative, in 10 per cent formalin or i

  18. Combined nitrogen limitation and cadmium stress stimulate total carbohydrates, lipids, protein and amino acid accumulation in Chlorella vulgaris (Trebouxiophyceae)

    Chia, Mathias Ahii, E-mail: [Department of Botany, Federal University of São Carlos, Rodovia Washington Luis km 235, São Carlos, SP Cep 13565905 (Brazil); Lombardi, Ana Teresa [Department of Botany, Federal University of São Carlos, Rodovia Washington Luis km 235, São Carlos, SP Cep 13565905 (Brazil); Graça Gama Melão, Maria da [Department of Hydrobiology, Federal University of São Carlos, Rodovia Washington Luis km 235, São Carlos, SP Cep 13565905 (Brazil); Parrish, Christopher C. [Department of Ocean Sciences, Memorial University of Newfoundland, St. John’s, Newfoundland A1C 5S7 (Canada)


    Highlights: • Chlorella vulgaris was exposed to Cd under varying N concentrations. • Growth rate and cell density decreased with increasing Cd stress and N limitation. • Dry weight, chlorophyll a, total lipid, carbohydrate and protein were accumulated. • Amino acids like proline and glutamine were accumulated under N and Cd stress. • Changes in amino acid composition are sensitive biomarkers for Cd and N stress. - Abstract: Metals have interactive effects on the uptake and metabolism of nutrients in microalgae. However, the effect of trace metal toxicity on amino acid composition of Chlorella vulgaris as a function of varying nitrogen concentrations is not known. In this research, C. vulgaris was used to investigate the influence of cadmium (10{sup −7} and 2.0 × 10{sup −8} mol L{sup −1} Cd) under varying nitrogen (2.9 × 10{sup −6}, 1.1 × 10{sup −5} and 1.1 × 10{sup −3} mol L{sup −1} N) concentrations on its growth rate, biomass and biochemical composition. Total carbohydrates, total proteins, total lipids, as well as individual amino acid proportions were determined. The combination of Cd stress and N limitation significantly inhibited growth rate and cell density of C. vulgaris. However, increasing N limitation and Cd stress stimulated higher dry weight and chlorophyll a production per cell. Furthermore, biomolecules like total proteins, carbohydrates and lipids increased with increasing N limitation and Cd stress. Ketogenic and glucogenic amino acids were accumulated under the stress conditions investigated in the present study. Amino acids involved in metal chelation like proline, histidine and glutamine were significantly increased after exposure to combined Cd stress and N limitation. We conclude that N limitation and Cd stress affects the physiology of C. vulgaris by not only decreasing its growth but also stimulating biomolecule production.

  19. A Systematic Analysis of Coal Accumulation Process

    CHENG Aiguo


    Formation of coal seam and coal-rich zone is an integrated result of a series of factors in coal accumulation process. The coal accumulation system is an architectural aggregation of coal accumulation factors. It can be classified into 4 levels: the global coal accumulation super-system, the coal accumulation domain mega.system, the coal accumulation basin system, and the coal seam or coal seam set sub-system. The coal accumulation process is an open, dynamic, and grey system, and is meanwhile a system with such natures as aggregation, relevance, entirety, purpose-orientated, hierarchy, and environment adaptability. In this paper, we take coal accumulation process as a system to study origin of coal seam and coal-rich zone; and we will discuss a methodology of the systematic analysis of coal accumulation process. As an example, the Ordos coal basin was investigated to elucidate the application of the method of the coal accumulation system analysis.

  20. Metal accumulating plants: Medium's role

    Rabier, J.; Prudent, P.; Szymanska, B.; Mevy, J.-P.


    To evaluate phytoremediation potentialities by metal accumulation in tolerant plants, trials are carried out using in vitro cultures. Organie compounds influence on metal accumulation is studied with metals supplemented media. The tested compounds on zinc and lead absorption by Brassica juncea, are chelating agents (EDTA, citric acid) and soluble organic fractions of compost. EDTA seems to enhance the transfer of lead in plant but it is the opposite in the case of zinc. Citric acid stimulates root absorption for both zinc and lead. For the aqueous extracts of compost, variable effects are obtained according to the origin of compost (green wastes and food wastes). In'all tested conditions of cultures, zinc is mainly exported towards shoot while lead is stored in root.

  1. Bacterial accumulation in viscosity gradients

    Waisbord, Nicolas; Guasto, Jeffrey


    Cell motility is greatly modified by fluid rheology. In particular, the physical environments in which cells function, are often characterized by gradients of viscous biopolymers, such as mucus and extracellular matrix, which impact processes ranging from reproduction to digestion to biofilm formation. To understand how spatial heterogeneity of fluid rheology affects the motility and transport of swimming cells, we use hydrogel microfluidic devices to generate viscosity gradients in a simple, polymeric, Newtonian fluid. Using video microscopy, we characterize the random walk motility patterns of model bacteria (Bacillus subtilis), showing that both wild-type ('run-and-tumble') cells and smooth-swimming mutants accumulate in the viscous region of the fluid. Through statistical analysis of individual cell trajectories and body kinematics in both homogeneous and heterogeneous viscous environments, we discriminate passive, physical effects from active sensing processes to explain the observed cell accumulation at the ensemble level.

  2. Mechanisms of intrahepatic triglyceride accumulation

    Ress, Claudia; Kaser, Susanne


    Hepatic steatosis defined as lipid accumulation in hepatocytes is very frequently found in adults and obese adolescents in the Western World. Etiologically, obesity and associated insulin resistance or excess alcohol intake are the most frequent causes of hepatic steatosis. However, steatosis also often occurs with chronic hepatitis C virus (HCV) infection and is also found in rare but potentially life-threatening liver diseases of pregnancy. Clinical significance and outcome of hepatic trigl...

  3. Mechanisms of intrahepatic triglyceride accumulation.

    Ress, Claudia; Kaser, Susanne


    Hepatic steatosis defined as lipid accumulation in hepatocytes is very frequently found in adults and obese adolescents in the Western World. Etiologically, obesity and associated insulin resistance or excess alcohol intake are the most frequent causes of hepatic steatosis. However, steatosis also often occurs with chronic hepatitis C virus (HCV) infection and is also found in rare but potentially life-threatening liver diseases of pregnancy. Clinical significance and outcome of hepatic triglyceride accumulation are highly dependent on etiology and histological pattern of steatosis. This review summarizes current concepts of pathophysiology of common causes of hepatic steatosis, including non-alcoholic fatty liver disease (NAFLD), alcoholic fatty liver disease, chronic HCV infections, drug-induced forms of hepatic steatosis, and acute fatty liver of pregnancy. Regarding the pathophysiology of NAFLD, this work focuses on the close correlation between insulin resistance and hepatic triglyceride accumulation, highlighting the potential harmful effects of systemic insulin resistance on hepatic metabolism of fatty acids on the one side and the role of lipid intermediates on insulin signalling on the other side. Current studies on lipid droplet morphogenesis have identified novel candidate proteins and enzymes in NAFLD.

  4. Fatigue Damage Accumulation of Steel/rubber Composite

    Zhenhui TIAN; Huifeng TAN; Xingwen DU


    The fatigue damage accumulation of [±20°] laminated steel cord reinforced rubber composite under T-T loading was studied. Results indicate that the increase in the cyclic maximum strain exhibits three-stage tendency in the process of fatigue. The macroscopic fatigue damage initiates from the ends of steel cords in the form of cylindrical crack. Damage propagates along with the increase in crack numbers, the cord/matrix interface debonding and the growth of interply cracks. By using the dynamic creep as parameter, a linear fatigue damage accumulation model was established. This model can be used under dual loading conditions to estimate the residual fatigue life of the specimen.

  5. Assessment of drug accumulation in the evaluation of pharmacokinetic data.

    Meineke, I; Gleiter, C H


    The evaluation of drug accumulation is approached from a practical point of view. Estimates of accumulation indices as obtained from standard estimators-AUC, peak levels, and trough levels (RAUAUC Rmax and Rmin, respectively)-are compared and differences analyzed. The estimators are based on the concentration-time curve characteristics area under the concentration-time curve (AUC), maximum concentration, and trough level. Simulated data are used for the analysis, both noise-free and with random error added. The data are based on pharmacokinetic parameters derived from a clinical study. The numerical procedures can be reproduced by the interested reader with little effort. It is shown empirically that if Rmin, > RAUC then simple kinetic behavior cannot be safely assumed, but accumulation is a complex function of time. Rmax as obtained from the data and an estimate of this value based on time to peak concentration (tmax) and apparent elimination rate constant (lambda(z)) after a single dose and at steady state can then be compared in an attempt to exclude time-dependent kinetics. This new numerical procedure can provide valuable and even pivotal information regarding the accumulation kinetics of a compound under investigation. Recommendations on how to use the available concentration-time information to the best advantage are presented. It is concluded that the assessment of drug accumulation should not be confined to the calculation of just one estimate, because the three estimators RAUC, Rmax. and Rmin reflect different aspects of accumulation. Moreover, all information about accumulation should be carefully analyzed in the clinical context. This way the relevant accumulation can be identified for safe and efficacious drug treatment.

  6. Kinetic parameters and tissue distribution of 5-oxo-L-prolinase determined by a fluorimetric assay.

    Weber, P; Jäger, M; Bangsow, T; Knell, G; Piechaczek, K; Koch, J; Wolf, S


    5-Oxo-L-prolinase (5-OPase) catalyses the hydrolysis of 5-oxo-L-proline to glutamate with concomitant stoichiometric cleavage of ATP to ADP, a reaction which is known to be part of the gamma-glutamyl cycle-an interrelated series of reactions involved in the synthesis and metabolism of glutathione. As recent studies indicate, this cyclic pathway plays a crucial role in the regulation of amino acid transport. Apparently, the intermediate product 5-oxo-L-proline functions as a second messenger molecule that upregulates the activity of certain amino acid transport systems. Thus, the degradation of 5-oxo-L-proline by 5-OPase leads to the downregulation of this stimulus. In this study, a new sensitive fluorimetric assay for 5-OPase activity was established which is based on the derivatization of glutamate with o-phthaldialdehyde in the presence of thiols and subsequent separation of the products by HPLC. The method is suitable for the screening of chromatography fractions as well as for the determination of the kinetic parameters Km and Vmax of purified 5-OPase. Additionally, it can be used for the measurement of enzyme activity in crude cell extracts and evaluation of tissue distribution.


    Cretu V.


    Full Text Available Based on theoretical and experimental research of the milk cooling process was elaborated mathematical model of accumulator with natural cold and vacuum installations, which allows determining the operating system installations, mentioned from environmental parameters.

  8. Effective Phytoextraction of Cadmium (Cd) with Increasing Concentration of Total Phenolics and Free Proline in Cannabis sativa (L) Plant Under Various Treatments of Fertilizers, Plant Growth Regulators and Sodium Salt.

    Ahmad, Ayaz; Hadi, Fazal; Ali, Nasir


    The comparative effect of fertilizers (NPK), plant growth regulators (GA3, IAA, Zeatin) and sodium chloride (NaCl) on Cd phytoaccumulation, proline and phenolics production in Cannabis sativa was evaluated. Proline and phenolices were correlated with Cd contents in plant. Cd significantly reduced the plant growth. Fertilizers application (in combination) most significantly increased the growth (19 cm root and 47 cm shoot) on Cd contaminated soil. All treatments increased the Cd contents in plant tissues. This increase was highly significant in fertilizers treated plants (1101, 121 and 544 ppm in roots, stem and leaves respectively). Significantly positive correlation was found between Cd concentration and dry biomass of root (R2=0.7511) and leaves (R2=0.5524). All treatments significantly increased the proline and total phenolics and maximum was recorded in NaCl treated plants followed by fertilizers. Proline was higher in roots while phenolics in leaves. The correlation between proline and phenolics was positive in leaf (R2=0.8439) and root (R2=0.5191). Proline and phenolics showed positive correlation with Cd concentration in plant. Conclusively, fertilizers in combination seem to be the better option for Cd phytoextraction. Further investigation is suggested to study the role of phenolics and proline in Cd phytoextraction.

  9. Dental wax decreases calculus accumulation in small dogs.

    Smith, Mark M; Smithson, Christopher W


    A dental wax was evaluated after unilateral application in 20 client-owned, mixed and purebred small dogs using a clean, split-mouth study model. All dogs had clinical signs of periodontal disease including plaque, calculus, and/or gingivitis. The wax was randomly applied to the teeth of one side of the mouth daily for 30-days while the contralateral side received no treatment. Owner parameters evaluated included compliance and a subjective assessment of ease of wax application. Gingivitis, plaque and calculus accumulation were scored at the end of the study period. Owners considered the wax easy to apply in all dogs. Compliance with no missed application days was achieved in 8 dogs. The number of missed application days had no effect on wax efficacy. There was no significant difference in gingivitis or plaque accumulation scores when comparing treated and untreated sides. Calculus accumulation scores were significantly less (22.1 %) for teeth receiving the dental wax.

  10. Evidence accumulation as a model for lexical selection.

    Anders, R; Riès, S; van Maanen, L; Alario, F X


    We propose and demonstrate evidence accumulation as a plausible theoretical and/or empirical model for the lexical selection process of lexical retrieval. A number of current psycholinguistic theories consider lexical selection as a process related to selecting a lexical target from a number of alternatives, which each have varying activations (or signal supports), that are largely resultant of an initial stimulus recognition. We thoroughly present a case for how such a process may be theoretically explained by the evidence accumulation paradigm, and we demonstrate how this paradigm can be directly related or combined with conventional psycholinguistic theory and their simulatory instantiations (generally, neural network models). Then with a demonstrative application on a large new real data set, we establish how the empirical evidence accumulation approach is able to provide parameter results that are informative to leading psycholinguistic theory, and that motivate future theoretical development. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Neurofilament Phosphorylation during Development and Disease: Which Came First, the Phosphorylation or the Accumulation?

    Jeffrey M. Dale


    Full Text Available Posttranslational modification of proteins is a ubiquitous cellular mechanism for regulating protein function. Some of the most heavily modified neuronal proteins are cytoskeletal proteins of long myelinated axons referred to as neurofilaments (NFs. NFs are type IV intermediate filaments (IFs that can be composed of four subunits, neurofilament heavy (NF-H, neurofilament medium (NF-M, neurofilament light (NF-L, and α-internexin. Within wild type axons, NFs are responsible for mediating radial growth, a process that determines axonal diameter. NFs are phosphorylated on highly conserved lysine-serine-proline (KSP repeats located along the C-termini of both NF-M and NF-H within myelinated axonal regions. Phosphorylation is thought to regulate aspects of NF transport and function. However, a key pathological hallmark of several neurodegenerative diseases is ectopic accumulation and phosphorylation of NFs. The goal of this review is to provide an overview of the posttranslational modifications that occur in both normal and diseased axons. We review evidence that challenges the role of KSP phosphorylation as essential for radial growth and suggests an alternative role for NF phosphorylation in myelinated axons. Furthermore, we demonstrate that regulation of NF phosphorylation dynamics may be essential to avoiding NF accumulations.

  12. Effect of Zn stresses on physiology, growth, Zn accumulation, and chlorophyll of Phyllostachys pubescen s.

    Peng, Danli; Shafi, Mohammad; Wang, Ying; Li, Song; Yan, Wenbo; Chen, Junren; Ye, Zhengqian; Liu, Dan


    The effects of Zinc (Zn) on lipid peroxidation, antioxidative enzymes, growth, Zn accumulation, and leaf chlorophyll of Phyllostachys pubescens (Pradelle) Mazel ex J.Houz. were investigated in two greenhouse experiments. Hydroponics experiment with Zn application of 0, 20, 100, and 400 μM revealed that lower concentration of Zn in solution led to increased malondialdehyde (MDA) and proline contents but inhibited SOD activity in all treatments. P. pubescens had showed strong ability to accumulate Zn in stems and reached maximum level at 100 μM with 7.91-fold increase compared with control. In pot experiment, treatment with Zn ranged from 0, 200, 400, 800, 1,600, to 3,200 mg kg(-1). Application of 800 mg kg(-1) revealed 116, 24.6, and 28.3 times increase in Zn concentration of roots, stems, and leaves, respectively. Growth and chlorophyll contents of plants in pots were better promoted at 400 mg kg(-1) Zn, with 60.5 and 30.9 % enhanced roots and shoot compared with control. The bioaccumulation factor (BAF) was in the sequence of stem > roots > leaves. The translocation factor (TF) of stem was higher than leaves.

  13. Low Temperature-Induced 30 (LTI30 positively regulates drought stress resistance in Arabidopsis: effect on abscisic acid sensitivity and hydrogen peroxide accumulation

    Haitao eShi


    Full Text Available As a dehydrin belonging to group II late embryogenesis abundant protein (LEA family, Arabidopsis Low Temperature-Induced 30 (LTI30/XERO2 has been shown to be involved in plant freezing stress resistance. However, the other roles of AtLTI30 remain unknown. In this study, we found that the expression of AtLTI30 was largely induced by drought stress and abscisic acid (ABA treatments. Thereafter, AtLTI30 knockout mutants and overexpressing plants were isolated to investigate the possible involvement of AtLTI30 in ABA and drought stress responses. AtLTI30 knockout mutants were less sensitive to ABA-mediated seed germination, while AtLTI30 overexpressing plants were more sensitive to ABA compared with wild type (WT. Consistently, the AtLTI30 knockout mutants displayed decreased drought stress resistance, while the AtLTI30 overexpressing plants showed improved drought stress resistance compared with WT, as evidenced by a higher survival rate and lower leaf water loss than WT after drought stress. Moreover, manipulation of AtLTI30 expression positively regulated the activities of catalases (CATs and endogenous proline content, as a result, negatively regulated drought stress-triggered hydrogen peroxide (H2O2 accumulation. All these results indicate that AtLTI30 is a positive regulator of plant drought stress resistance, partially through the modulation of ABA sensitivity, H2O2 and proline accumulation.

  14. Tipping news in information accumulation system

    Shin, J. K.


    As a continuous opinion dynamics model, the information accumulation system (IAS) includes three basic mechanisms of the news, the inheritance and the diffusion as contributing to the information accumulation process of a system. A system is composed of agents who diffuse information through internal interaction, while each of them has incomplete memory or inheritance rate. The news comes from external sources of information, such as mass media. Previously the model IAS was studied only for the small news problems. In this study, a tipping news problem is considered. A key question of the problem is: what is the minimum strength of advertisement that can tip the minority opinion to a majority one? Dynamics of the IAS is briefly revisited with a special interest on nonlinear behavior of the model. In particular, it is shown that a discrete map of the IAS for a single color problem can be transformed into a logistic map, from which the dynamics of the IAS can be better understood. To show the applicability of the IAS model, the result is applied to explain the concept of the critical population size, which claims that there is a minimum population size for a social knowledge system to be continuously inherited without being lost. And critical size of the tipping news is found analytically in terms of IAS parameters. Some of the key results from the present study are compared in detail with the results from the Brownian particle model, which is believed to be the most similar model to the IAS. The concept of tipping news is used to show that a traditional society can tip at an exceptionally low inter-community exposure. Finally, the result was applied to the language competition problem.

  15. Solution structure of the human BTK SH3 domain complexed with a proline-rich peptide from p120cbl

    Tzeng, S.-R.; Lou, Y.-C.; Pai, M.-T.; Jain, Moti L.; Cheng, J.-W. [National Tsing Hua University, Division of Structural Biology and Biomedical Science, Department of Life Science (China)


    X-linked agammaglobulinemia (XLA), an inherited disease, is caused by mutations in the Bruton's tyrosine kinase (BTK). The absence of functional BTK leads to failure of B cell differentiation which incapacitates antibody production in XLA patients leading to, sometimes lethal, bacterial infections. Point mutation in the BTK gene that leads to deletion of C-terminal 14 aa residues of BTK SH3 domain was found in one patient family. To understand the role of BTK in B cell development, we have determined the solution structure of BTK SH3 domain complexed with a proline-rich peptide from the protein product of c-cbl protooncogene (p120{sup cbl}). Like other SH3 domains, BTK SH3 domain consists of five {beta}-strands packed in two {beta}-sheets forming a {beta}-barrel-like structure. The rmsd calculated from the averaged coordinates for the BTK SH3 domain residues 218-271 and the p120{sup cbl} peptide residues 6-12 of the complex was 0.87 A ({+-}0.16 A) for the backbone heavy atoms (N, C, and C{sub {alpha}}) and 1.64 A ({+-}0.16 A) for all heavy atoms. Based on chemical shift changes and inter-molecular NOEs, we have found that the residues located in the RT loop, n-Src loop and helix-like loop between {beta}4 and {beta}5 of BTK SH3 domain are involved in ligand binding. We have also determined that the proline-rich peptide from p120{sup cbl} binds to BTK SH3 domain in a class I orientation. These results correlate well with our earlier observation that the truncated BTK SH3 domain (deletion of {beta}4, {beta}5 and the helix-like loop) exhibits weaker affinity for the p120{sup cbl} peptide. It is likely that the truncated SH3 domain fails to present to the ligand the crucial residues in the correct context and hence the weaker binding. These results delineate the importance of the C-terminus in the binding of SH3 domains and also indicate that improper folding and the altered binding behavior of mutant BTK SH3 domain likely lead to XLA.

  16. Human dipeptidyl peptidase III acts as a post-proline-cleaving enzyme on endomorphins.

    Barsun, Marina; Jajcanin, Nina; Vukelić, Bojana; Spoljarić, Jasminka; Abramić, Marija


    Dipeptidyl peptidase III (DPP III) is a zinc exopeptidase with an implied role in the mammalian pain-modulatory system owing to its high affinity for enkephalins and localisation in the superficial laminae of the spinal cord dorsal horn. Our study revealed that this human enzyme hydrolyses opioid peptides belonging to three new groups, endomorphins, hemorphins and exorphins. The enzymatic hydrolysis products of endomorphin-1 were separated and quantified by capillary electrophoresis and the kinetic parameters were determined for human DPP III and rat DPP IV. Both peptidases cleave endomorphin-1 at comparable rates, with liberation of the N-terminal Tyr-Pro. This is the first evidence of DPP III acting as an endomorphin-cleaving enzyme.

  17. Biota-Sediment Accumulation Factor Data

    U.S. Environmental Protection Agency — The Biota-Sediment Accumulation Factor contains approximately 20,000 biota-sediment accumulation factors (BSAFs) from 20 locations (mostly Superfund sites) for...

  18. Lead Tolerance and Accumulation in White Poplar Cultivated In Vitro

    Branislav Kovačević


    Full Text Available Background and Purpose: This paper analyses the lead tolerance and accumulation in white poplar genotypes in vitro, in order to optimize genotype evaluation and other procedures in their implementation in phytoremediation projects and landscaping in areas endangered by lead accumulation. Material and Methods: The lead tolerance and accumulation of five white poplar genotypes after 35 days in vitro cultivation on media supplemented with lead was examined. The following Pb(NO32 concentrations were used: 0, 10-6, 10-5, 10-4 and 10-3 M. Tolerance analysis (described by tolerance indices was based on morphological parameters, biomass accumulation and the content of photosynthetic pigments, while lead accumulation was described by shoot lead accumulation and shoot lead content. Results and Conclusions: The chosen lead concentrations appeared not to be lethal. Moreover, the obtained results showed that the tested lead concentrations had a positive effect on: number of formed roots, shoot moisture content and shoot height. The best differentiation among the examined genotypes was gained by the tolerance index based on the shoot height on 10-4 M Pb(NO32. The shoot lead accumulation and shoot lead content significantly increased on 10-4 and 10-3 M Pb(NO32 media. Thus, the concentration of 10-4 M Pb(NO32 is recommended for further research. Two examined genotypes of horticultural value (LCM and LBM achieved a significantly higher lead shoot content compared to the wide spread genotype “Villafranca” (almost 200% and 125% higher, respectively.

  19. Technical Benefits of Tram Vehicle Energy Accumulator

    Zbigniew Drazek


    Full Text Available Energy saving effects connected with use of energy accumulator on board of tram vehicle instead of substation are presented. Differences in results regarding weak and strong power supply system when taking into account energy losses and energy recuperation are pointed out. Running mode and energy changed from substation by a tram vehicle with accumulator is compared to a tram vehicle without on-board accumulator but supplied from substation equipped with energy accumulator.

  20. 47 CFR 32.3100 - Accumulated depreciation.


    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Accumulated depreciation. 32.3100 Section 32... Accumulated depreciation. (a) This account shall include the accumulated depreciation associated with the... with depreciation amounts concurrently charged to Account 6561, Depreciation...

  1. Models to estimate phytomass accumulation of hydroponic lettuce

    Lopes,Sidinei José; Dourado Neto,Durval; MANFRON, Paulo Augusto; Jasniewicz,Luís Renato


    The protected and hydroponics cultivation are increasing in Brazil, demanding a better knowledge of crop performance in this environment. Plant dry matter accumulation as a function of solar radiation, temperature, relative humidity and other weather parameters in greenhouse is different when compared with field cultivation. With the purpose of proposing models to characterize the temporal variation of leaf and total dry matter production of hydroponics-grown lettuce (Lactuca sativa L.) and t...

  2. Mutation accumulation and the catastrophic senescence of Pacific salmon

    Penna, T J P; Stauffer, D; Stauffer, Dietrich


    The bit-string model of biological aging is used to simulate the catastrophic senescence of Pacific Salmon. We have shown that reproduction occuring only once and at a fixed age is the only ingredient needed to explain the catastrophic senescence according the mutation accumulation theory. Several results are presented, some of them with up to 10^8 fishes, showing how the survival rates in catastrophic senescence are affected by changes in the parameters of the model.

  3. Electron-Positron Accumulator (EPA)

    Photographic Service


    After acceleration in the low-current linac LIL-W, the electrons and positrons are accumulated in EPA to obtain a sufficient intensity and a suitable time-structure, before being passed on to the PS for further acceleration to 3.5 GeV. Electrons circulate from right to left, positrons in the other direction. Dipole bending magnets are red, focusing quadrupoles blue, sextupoles for chromaticity-control orange. The vertical tube at the left of the picture belongs to an optical transport system carrying the synchrotron radiation to detectors for beam size measurement. Construction of EPA was completed in spring 1986. LIL-W and EPA were conceived for an energy of 600 MeV, but operation was limited to 500 MeV.

  4. Energy Accumulation by Hydrogen Technologies

    Jiřina Čermáková


    Full Text Available Photovoltaic power plants as a renewable energy source have been receiving rapidly growing attention in the Czech Republic and in the other EU countries. This rapid development of photovoltaic sources is having a negative effect on the electricity power system control, because they depend on the weather conditions and provide a variable and unreliable supply of electric power. One way to reduce this effect is by accumulating electricity in hydrogen. The aim of this paper is to introduce hydrogen as a tool for regulating photovoltaic energy in island mode. A configuration has been designed for connecting households with the photovoltaic hybrid system, and a simulation model has been made in order to check the validity of this system. The simulation results provide energy flows and have been used for optimal sizing of real devices. An appropriate system can deliver energy in a stand-alone installation.

  5. An amperometric D-amino acid biosensor prepared with a thermostable D-proline dehydrogenase and a carbon nanotube-ionic liquid gel.

    Tani, Yuji; Itoyama, Yukiko; Nishi, Kenichi; Wada, Chikahiro; Shoda, Yoshio; Satomura, Takenori; Sakuraba, Haruhiko; Ohshima, Toshihisa; Hayashi, Yukako; Yabutani, Tomoki; Motonaka, Junko


    Carbon nanotube (CNT) gel, which is composed of a mixture of single-wall CNT, an ionic liquid, and a thermostable D-proline dehydrogenase (D-Pro DH) immobilized electrode was utilized for the determination of D-amino acids (DAAs) in food samples. When a critical comparison with CNT, Ketjen Black (KB), and carbon powder (CP) was also carried out, the CNT/D-Pro DH immobilized electrode showed the highest sensitivity and the lowest detection limit of D-proline. In addition, the CNT/D-Pro DH immobilized electrode was applied to detection of DAAs in rice wine and vinegar samples. The concentrations of DAAs in rice wine and vinegar samples were 0.0210 +/- 0.0001 and 0.55 +/- 0.05 mmol L(-1), respectively.

  6. Study of the effects of proline, phenylalanine, and urea foliar application to Tempranillo vineyards on grape amino acid content. Comparison with commercial nitrogen fertilisers.

    Garde-Cerdán, T; López, R; Portu, J; González-Arenzana, L; López-Alfaro, I; Santamaría, P


    The aim of this work was to study the influence of foliar application of different nitrogen sources on grape amino acid content. The nitrogen sources applied to Tempranillo grapevines were proline, phenylalanine, urea, and two commercial nitrogen fertilisers, both without and with amino acids in their formulations. All treatments were applied at veraison and one week later. Proline treatment did not affect the must nitrogen composition. However, phenylalanine and urea foliar application enhanced the plants' synthesis of most of the amino acids, producing similar effects. In addition, the spray of commercial nitrogen fertilisers over leaves also induced a rise in grape amino acid concentrations regardless of the presence or absence of amino acids in their formulation. The most effective treatments were phenylalanine and urea followed by nitrogen fertilisers. This finding is of oenological interest for improved must nitrogen composition, ensuring better fermentation kinetics and most likely enhancing wine quality.

  7. Association of the leucine-7 to proline-7 variation in the signal sequence of neuropeptide Y with major depression.

    Koefoed, Pernille; Woldbye, David P D; Hansen, Thomas V O; Eplov, Lene F; Christiansen, Søren H; Mors, Ole; Kessing, Lars V; Werge, Thomas; Kaipio, Katja; Pesonen, Ullamari; Fahmy, Thomas; Mellerup, Erling; Jakobsen, Klaus D; Hansen, Elsebeth S; Knudsen, Gitte M; Bukh, Jens D; Bock, Camilla; Lindberg, Camilla; Kristensen, Ann S; Dam, Henrik; Nordentoft, Merete; Als, Thomas D; Wang, August G; Gether, Ulrik; Rehfeld, Jens F; Bolwig, Tom G


    There is clear evidence of a genetic component in major depression, and several studies indicate that neuropeptide Y (NPY) could play an important role in the pathophysiology of the disease. A well-known polymorphism encoding the substitution of leucine to proline in the signal peptide sequence of NPY (Leu7Pro variation) was previously found to protect against depression. Our study aimed at replicating this association in a large Danish population with major depression. Leu7Pro was studied in a sample of depressed patients and ethnically matched controls, as well as psychiatric disease controls with schizophrenia. Possible functional consequences of Leu7Pro were explored in vitro. In contrast to previous studies, Pro7 appeared to be a risk allele for depression, being significantly more frequent in the depression sample (5.5%, n = 593; p = 0.009; odds ratio, OR: 1.46) as compared to ethnically matched controls (3.8%, n = 2912), while schizophrenia patients (4.1%, n = 503) did not differ. In vitro, the Pro7 substitution appeared to be associated with reduced levels of NPY without affecting its mRNA level. The Leu7Pro variation may increase the risk of major depression, possibly by affecting the biosynthesis of NPY.

  8. Enhanced endothelium-dependent vasodilation in subjects with Proline7 substitution in the signal peptide of neuropeptide Y.

    Järvisalo, Mikko J; Jartti, Laura; Karvonen, Matti K; Pesonen, Ullamari; Koulu, Markku; Marniemi, Jukka; Hammar, Niklas; Kaprio, Jaakko; Paakkunainen, Ulla; Simell, Olli; Raitakari, Olli T; Rönnemaa, Tapani


    Neuropeptide Y (NPY) is a molecule that may have both vasoconstrictive and vasodilatory actions. A common polymorphism in the human NPY gene that results in the Leucine7 to Proline7 substitution (Leu7Pro) in the signal peptide part of the NPY was recently identified. This substitution has been associated with elevated serum cholesterol levels and with slightly accelerated progression rate of carotid intima-media thickness, thus suggesting increased risk of atherosclerosis in carriers of Pro7 substitution. Recent data also indicate that subjects with Pro7 substitution may have increased endothelial release of NPY. This study was undertaken to elucidate the effects of Leu7Pro polymorphism on arterial endothelial function. We measured flow-mediated endothelial-dependent dilatation (FMD) of the brachial artery in two separate populations: in 152 middle-aged men and in 95 prepubertal children. In both study populations, subjects with Pro7 substitution had 48-52% higher FMD compared with subjects having the wildtype (Leu7/Leu7) signal peptide sequence. We conclude that Pro7 substitution in signal peptide of the NPY is associated with enhanced endothelial-dependent vasodilation. Prospective studies are needed to determine whether Pro7 substitution is associated with increased or decreased risk of cardiovascular morbidity and mortality.

  9. Synthesis of deoxynucleoside triphosphates that include proline, urea, or sulfonamide groups and their polymerase incorporation into DNA.

    Hollenstein, Marcel


    To expand the chemical array available for DNA sequences in the context of in vitro selection, I present herein the synthesis of five nucleoside triphosphate analogues containing side chains capable of organocatalysis. The synthesis involved the coupling of L-proline-containing residues (dU(tP)TP and dU(cP)TP), a dipeptide (dU(FP)TP), a urea derivative (dU(Bpu)TP), and a sulfamide residue (dU(Bs)TP) to a suitably protected common intermediate, followed by triphosphorylation. These modified dNTPs were shown to be excellent substrates for the Vent (exo(-)) and Pwo DNA polymerases, as well as the Klenow fragment of E. coli DNA polymerase I, although they were only acceptable substrates for the 9°N(m) polymerase. All of the modified dNTPs, with the exception of dU(Bpu)TP, were readily incorporated into DNA by the polymerase chain reaction (PCR). Modified oligonucleotides efficiently served as templates for PCR for the regeneration of unmodified DNA. Thermal denaturation experiments showed that these modifications are tolerated in the major groove. Overall, these heavily modified dNTPs are excellent candidates for SELEX.

  10. Binding affinity of hydrolyzable tannins to parotid saliva and to proline-rich proteins derived from it.

    Bacon, J R; Rhodes, M J


    Proline-rich proteins (PRP) in human parotid saliva have a high affinity for dietary polyphenolic compounds (tannins), forming stable complexes that may modulate the biological and nutritional properties of the tannin. The formation of such complexes may also have an important role in the modulation or promotion of the sensation of oral astringency perceived when tannin-rich foods and beverages are consumed. The major classes of PRP (acidic, basic, and glycosylated) have been isolated from human saliva, and the relative binding affinities of a series of hydrolyzable tannins, which are found in a number of plant-derived foods and beverages, to these PRP classes have been determined using a competition assay. All of the classes of PRP have a high capacity for hydrolyzable tannins. Within the narrow range of binding affinities exhibited, structure/binding relationships with the levels of tannin galloylation, hexahydroxydiphenoyl esterification, and degree of polymerization were identified. No individual class of human salivary PRP appears to have an exclusive affinity for a particular type of hydrolyzable tannin.

  11. Ethanol Concentration Influences the Mechanisms of Wine Tannin Interactions with Poly(L-proline) in Model Wine.

    McRae, Jacqui M; Ziora, Zyta M; Kassara, Stella; Cooper, Matthew A; Smith, Paul A


    Changes in ethanol concentration influence red wine astringency, and yet the effect of ethanol on wine tannin-salivary protein interactions is not well understood. Isothermal titration calorimetry (ITC) was used to measure the binding strength between the model salivary protein, poly(L-proline) (PLP) and a range of wine tannins (tannin fractions from a 3- and a 7-year old Cabernet Sauvignon wine) across different ethanol concentrations (5, 10, 15, and 40% v/v). Tannin-PLP interactions were stronger at 5% ethanol than at 40% ethanol. The mechanism of interaction changed for most tannin samples across the wine-like ethanol range (10-15%) from a combination of hydrophobic and hydrogen binding at 10% ethanol to only hydrogen binding at 15% ethanol. These results indicate that ethanol concentration can influence the mechanisms of wine tannin-protein interactions and that the previously reported decrease in wine astringency with increasing alcohol may, in part, relate to a decrease tannin-protein interaction strength.

  12. Lacrimal proline rich 4 (LPRR4 protein in the tear fluid is a potential biomarker of dry eye syndrome.

    Saijyothi Venkata Aluru

    Full Text Available Dry eye syndrome (DES is a complex, multifactorial, immune-associated disorder of the tear and ocular surface. DES with a high prevalence world over needs identification of potential biomarkers so as to understand not only the disease mechanism but also to identify drug targets. In this study we looked for differentially expressed proteins in tear samples of DES to arrive at characteristic biomarkers. As part of a prospective case-control study, tear specimen were collected using Schirmer strips from 129 dry eye cases and 73 age matched controls. 2D electrophoresis (2DE and Differential gel electrophoresis (DIGE was done to identify differentially expressed proteins. One of the differentially expressed protein in DES is lacrimal proline rich 4 protein (LPRR4. LPRR4 protein expression was quantified by enzyme immune sorbent assay (ELISA. LPRR4 was down regulated significantly in all types of dry eye cases, correlating with the disease severity as measured by clinical investigations. Further characterization of the protein is required to assess its therapeutic potential in DES.

  13. Detection of proline-rich proteins for the identification of saliva by enzyme-linked immunosorbent assay.

    Igoh, Akihisa; Tomotake, Sho; Doi, Yusuke


    Saliva is one of the most common body fluids found at a crime scene. Therefore, identifying saliva is important in forensic science. However, the current protein marker assays used to identify saliva are not sufficiently specific. Although proline-rich proteins (PRPs) are highly specific for saliva, their forensic potential has not yet been investigated. In this study, we developed enzyme-linked immunosorbent assays (ELISAs) to detect acidic salivary PRP HaeIII subfamily 1/2 (PRH1/2) and basic salivary PRP 2 (PRB2). The specificity, sensitivity, and efficiency of the ELISAs for PRH1/2 and PRB2 were compared with those of the ELISA for statherin (STATH), a known protein marker for saliva. The levels of PRH1/2 were significantly higher in saliva and saliva stains than in other body fluids (nasal secretions, urine, semen, vaginal fluid, blood, and sweat). PRB2 and STATH were detected in both nasal secretions and saliva. The PRH1/2 ELISA showed sensitivity similar to that of STATH ELISA. The detection rate of PRH1/2 ELISA was almost similar to that of STATH ELISA, followed by the ELISA for PRB2. The PRH1/2 ELISA had higher specificity for saliva than STATH ELISA. Therefore, the PRH1/2 ELISA has potential as a method to identify saliva for forensic investigation.

  14. Perspective of future drugs targeting sterile 20/SPS1-related proline/alanine-rich kinase for blood pressure control

    Lin, Gen-Min; Liu, Pang-Yen; Wu, Ching-Fen; Wang, Wen-Been; Han, Chih-Lu


    According to a genome-wide association study, intronic SNPs within the human sterile 20/SPS1-related proline/alanine-rich kinase (SPAK) gene was linked to 20% of the general population and may be associated with elevated blood pressure. As cell volume changes, mammalian SPAK kinases respond to phosphorylate and regulate cation-coupled chloride co-transporter activity. To our knowledge, phosphorylation of upstream with-no-lysine (K) (WNK) kinases would activate SPAK kinases. The activation of WNK-OSR1/SPAK cascade on the kidneys and aortic tissue is related to the development of hypertension. Several regulators of the WNK pathway such as the Kelch kinase protein 3 - Cullin 3 E3 ligase, hyperinsulinemia, and low potassium intake to mediate hypertension have been identified. In addition, the SPAK kinases may affect the action of renin-angiotensin-aldosterone system on blood pressure as well. In 2010, two SPAK knock-in and knock-out mouse models have clarified the pathogenesis of lowering blood pressure by influencing the receptors on the kidneys and aortic smooth muscle. More recently, two novel SPAK inhibitors for mice, Stock 1S-14279 and Closantel were discovered in 2014. Targeting of SPAK seems to be promising for future antihypertensive therapy. Therefore we raised some viewpoints for the issue for the antihypertensive therapy on the SPAK (gene or kinase). PMID:26131334

  15. Perspective of future drugs targeting sterile 20/SPS1-related proline/alanine-rich kinase for blood pressure control

    Gen-Min; Lin; Pang-Yen; Liu; Ching-Fen; Wu; Wen-Been; Wang; Chih-Lu; Han


    According to a genome-wide association study,intronic SNPs within the human sterile 20/SPS1-related proline/alanine-rich kinase(SPAK) gene was linked to 20% of the general population and may be associated with elevated blood pressure. As cell volume changes,mammalian SPAK kinases respond to phosphorylate and regulate cation-coupled chloride co-transporter activity. To our knowledge,phosphorylation of upstream with-no-lysine(K)(WNK) kinases would activate SPAK kinases. The activation of WNK-OSR1/SPAK cascade on the kidneys and aortic tissue is related to the development of hypertension. Several regulators of the WNK pathway such as the Kelch kinase protein 3-Cullin 3 E3 ligase,hyperinsulinemia,and low potassium intake to mediate hypertension have been identified. In addition,the SPAK kinases may affect the action of renin-angiotensin-aldosterone system on blood pressure as well. In 2010,two SPAK knock-in and knock-out mouse models have clarified the pathogenesis of lowering blood pressure by influencing the receptors on the kidneys and aortic smooth muscle. More recently,two novel SPAK inhibitors for mice,Stock 1S-14279 and Closantel were discovered in 2014. Targeting of SPAK seems to be promising for future antihypertensive therapy. Therefore we raised some viewpoints for the issue for the antihypertensive therapy on the SPAK(gene or kinase).

  16. RNA sequencing and pathway analysis identify tumor necrosis factor alpha driven small proline-rich protein dysregulation in chronic rhinosinusitis.

    Ramakrishnan, Vijay R; Gonzalez, Joseph R; Cooper, Sarah E; Barham, Henry P; Anderson, Catherine B; Larson, Eric D; Cool, Carlyne D; Diller, John D; Jones, Kenneth; Kinnamon, Sue C


    Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory disorder in which many pathways contribute to end-organ disease. Small proline-rich proteins (SPRR) are polypeptides that have recently been shown to contribute to epithelial biomechanical properties relevant in T-helper type 2 inflammation. There is evidence that genetic polymorphism in SPRR genes may predict the development of asthma in children with atopy and, correlatively, that expression of SPRRs is increased under allergic conditions, which leads to epithelial barrier dysfunction in atopic disease. RNAs from uncinate tissue specimens from patients with CRS and control subjects were compared by RNA sequencing by using Ingenuity Pathway Analysis (n = 4 each), and quantitative polymerase chain reaction (PCR) (n = 15). A separate cohort of archived sinus tissue was examined by immunohistochemistry (n = 19). A statistically significant increase of SPRR expression in CRS sinus tissue was identified that was not a result of atopic presence. SPRR1 and SPRR2A expressions were markedly increased in patients with CRS (p < 0.01) on RNA sequencing, with confirmation by using real-time PCR. Immunohistochemistry of archived surgical samples demonstrated staining of SPRR proteins within squamous epithelium of both groups. Pathway analysis indicated tumor necrosis factor (TNF) alpha as a master regulator of the SPRR gene products. Expression of SPRR1 and of SPRR2A is increased in mucosal samples from patients with CRS and appeared as a downstream result of TNF alpha modulation, which possibly resulted in epithelial barrier dysfunction.

  17. Thermodynamic contribution of backbone conformational entropy in the binding between SH3 domain and proline-rich motif.

    Zeng, Danyun; Shen, Qingliang; Cho, Jae-Hyun


    Biological functions of intrinsically disordered proteins (IDPs), and proteins containing intrinsically disordered regions (IDRs) are often mediated by short linear motifs, like proline-rich motifs (PRMs). Upon binding to their target proteins, IDPs undergo a disorder-to-order transition which is accompanied by a large conformational entropy penalty. Hence, the molecular mechanisms underlying control of conformational entropy are critical for understanding the binding affinity and selectivity of IDPs-mediated protein-protein interactions (PPIs). Here, we investigated the backbone conformational entropy change accompanied by binding of the N-terminal SH3 domain (nSH3) of CrkII and PRM derived from guanine nucleotide exchange factor 1 (C3G). In particular, we focused on the estimation of conformational entropy change of disordered PRM upon binding to the nSH3 domain. Quantitative characterization of conformational dynamics of disordered peptides like PRMs is limited. Hence, we combined various methods, including NMR model-free analysis, δ2D, DynaMine, and structure-based calculation of entropy loss. This study demonstrates that the contribution of backbone conformational entropy change is significant in the PPIs mediated by IDPs/IDRs.

  18. A cis-proline in alpha-hemoglobin stabilizing protein directs the structural reorganization of alpha-hemoglobin.

    Gell, David A; Feng, Liang; Zhou, Suiping; Jeffrey, Philip D; Bendak, Katerina; Gow, Andrew; Weiss, Mitchell J; Shi, Yigong; Mackay, Joel P


    alpha-Hemoglobin (alphaHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with alphaHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting alphaHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in alphaHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro(30), in loop 1 of AHSP. Mutation of Pro(30) to a variety of residue types results in reduced ability to convert alphaHb. In complex with alphaHb, AHSP Pro(30) adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in alphaHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the alphaHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of alphaHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops.

  19. A cis-Proline in α-Hemoglobin Stabilizing Protein Directs the Structural Reorganization of α-Hemoglobin*

    Gell, David A.; Feng, Liang; Zhou, Suiping; Jeffrey, Philip D.; Bendak, Katerina; Gow, Andrew; Weiss, Mitchell J.; Shi, Yigong; Mackay, Joel P.


    α-Hemoglobin (αHb) stabilizing protein (AHSP) is expressed in erythropoietic tissues as an accessory factor in hemoglobin synthesis. AHSP forms a specific complex with αHb and suppresses the heme-catalyzed evolution of reactive oxygen species by converting αHb to a conformation in which the heme is coordinated at both axial positions by histidine side chains (bis-histidyl coordination). Currently, the detailed mechanism by which AHSP induces structural changes in αHb has not been determined. Here, we present x-ray crystallography, NMR spectroscopy, and mutagenesis data that identify, for the first time, the importance of an evolutionarily conserved proline, Pro30, in loop 1 of AHSP. Mutation of Pro30 to a variety of residue types results in reduced ability to convert αHb. In complex with αHb, AHSP Pro30 adopts a cis-peptidyl conformation and makes contact with the N terminus of helix G in αHb. Mutations that stabilize the cis-peptidyl conformation of free AHSP, also enhance the αHb conversion activity. These findings suggest that AHSP loop 1 can transmit structural changes to the heme pocket of αHb, and, more generally, highlight the importance of cis-peptidyl prolyl residues in defining the conformation of regulatory protein loops. PMID:19706593

  20. A Linear Surface Epitope in a Proline-Rich Region of ORF3 Product of Genotype 1 Hepatitis E Virus

    Yang, Yonglin; Lin, Shaoli; Nan, Yuchen; Ma, Zexu; Yang, Liping; Zhang, Yanjin


    Hepatitis E virus (HEV) is one of the viral pathogens causing hepatitis in humans. HEV open reading frame 3 (ORF3) encodes a small multifunctional protein (VP13), which is essential for HEV infection. In this study, a linear epitope was identified in a polyproline (PXXP) motif from VP13 of genotype 1 HEV by using a monoclonal antibody. The epitope was detected in enzyme-linked immunosorbent assay (ELISA), immunoblotting and immunofluorescence assays. Epitope mapping showed that the epitope locates in a proline-rich region containing a PXXP motif in amino acid residues 66-75 of VP13. The epitope was also detected in HEV-infected liver cells and reacted with genotype 1-specific antibodies in an HEV-positive human serum sample. The results demonstrated that the epitope in the PXXP motif of the genotype 1 VP13 is linear and surface-oriented, which should facilitate in-depth studies on the viral protein and HEV biology. PMID:27548202

  1. A Linear Surface Epitope in a Proline-Rich Region of ORF3 Product of Genotype 1 Hepatitis E Virus

    Yonglin Yang


    Full Text Available Hepatitis E virus (HEV is one of the viral pathogens causing hepatitis in humans. HEV open reading frame 3 (ORF3 encodes a small multifunctional protein (VP13, which is essential for HEV infection. In this study, a linear epitope was identified in a polyproline (PXXP motif from VP13 of genotype 1 HEV by using a monoclonal antibody. The epitope was detected in enzyme-linked immunosorbent assay (ELISA, immunoblotting and immunofluorescence assays. Epitope mapping showed that the epitope locates in a proline-rich region containing a PXXP motif in amino acid residues 66-75 of VP13. The epitope was also detected in HEV-infected liver cells and reacted with genotype 1-specific antibodies in an HEV-positive human serum sample. The results demonstrated that the epitope in the PXXP motif of the genotype 1 VP13 is linear and surface-oriented, which should facilitate in-depth studies on the viral protein and HEV biology.

  2. Copper(II) interaction with peptide fragments of histidine-proline-rich glycoprotein: Speciation, stability and binding details.

    La Mendola, Diego; Magrì, Antonio; Santoro, Anna Maria; Nicoletti, Vincenzo G; Rizzarelli, Enrico


    GHHPH is the peptide repeat present in histidine-proline rich glycoprotein (HPRG), a plasma glycoprotein involved in angiogenesis process. The copper(II) ions interaction with mono (Ac-GHHPHG-NH(2)) and its bis-repeat (Ac-GHHPHGHHPHG-NH(2)) was investigated by means of potentiometric and spectroscopic techniques. To single out the copper(II) coordination environments of different species formed with Ac-GHHPHG-NH(2), three single point mutated peptides were also synthesized and their ability to coordinate Cu(2+) investigated. Ac-GHHPHG-NH(2) binds Cu(2+) by the imidazole side chain and the amide nitrogen deprotonation that takes place towards the N-terminus. The bis-repeat is able to bind Cu(2+) more efficiently than Ac-GHHPHG-NH(2). This difference is not only due to the number of His residues in the sequence but also to the different binding sites. In fact, the comparison of the potentiometric and spectroscopic data of the copper(II) complexes with a bis-repeatPeg construct Ac-(GHHPHG)-Peg-(GHHPHG)-NH(2) and those of the metal complexes with Ac-HGHH-NH(2), indicates that the central HGHH amino acid sequence is the main copper(II) binding site.

  3. Site-specific indolation of proline-based peptides via copper(II)-catalyzed oxidative coupling of tertiary amine N-oxides.

    Wu, Xiaowei; Zhang, Dengyou; Zhou, Shengbin; Gao, Feng; Liu, Hong


    The first site-specific and purely chemical method for modifying proline-based peptides was developed via a convenient, copper-catalyzed oxidative coupling of tertiary amine N-oxides with indoles. This novel approach features high regioselectivity and diastereoselectivity, mild conditions, and compatibility with various functional groups. In addition, a simplified process was realized in one pot and two steps via in situ oxidative coupling of tertiary amine and indoles.

  4. Varying light regimes in naturally growing Jatropha curcus: pigment, proline and photosynthetic performance

    Wadhwa R.


    Full Text Available Light stress is a major abiotic stress which adversely affects productivity of the plants. Tolerance to abiotic stresses is very complex, due to the intricate of interactions between stress factors and various molecular, biochemical and physiological phenomena affecting plant growth and development. In many cases, high yield potential can contribute to yield in moderate stress environment. We studied chlorophyll (Chl a fluorescence parameters and analyzed D1 core protein in one year old plants of Jatropha curcus under different light regimes (10–1200 μmol m–2 s–1 in sun and shade plants. Chl a fluorescence provides insights into the responses of the photosynthetic system to increasing irradiance. Total Chl content was 1.43 and 0.61 mg/g-1 FM for shade and sun exposed plants respectively. The effective quantum yield (ΔF/Fm' of the sun plants was lower as compared to shade plants but the amount of the D1 core protein was higher in plants grown under high light intensity. A decrease in ΔF/Fm' indicates down regulation of photosynthesis or photoinhibition. D1 protein is the membrane protein complex of the PSII reaction centre. The degradation of D1 protein may regulate the functioning of the PSII repair cycle under photoinhibitory conditions. It has been shown that low-light grown or shade plants are more susceptible to photoinhibition than high light or sun plants. This higher susceptibility is accompanied by slow degradation of damaged D1 protein. High light intensity or exposure to photooxidation leads to the irreversible damage in photosynthetic performance and consequently has an overall inhibitory effect on crop productivity.

  5. Impacts of proline on the central metabolism of an industrial erythromycin-producing strain Saccharopolyspora erythraea via (13)C labeling experiments.

    Hong, Ming; Huang, Mingzhi; Chu, Ju; Zhuang, Yingping; Zhang, Siliang


    Saccharopolyspora erythraea E3 is an important industrial strain for erythromycin production and knowledge on its metabolism is limited. In the present work, (13)C labeling experiments were conducted to characterize the metabolism of S. erythraea E3. We found that S. erythraea E3 was difficult to grow on minimal medium with glucose as sole carbon source and the addition of proline remarkably improved the cell growth. The activity of EMP pathway was very low and ED pathway was alternatively the main glucose utilization pathway. The addition of proline resulted in remarkable changes in the fluxes of central metabolism. The fluxes in PP pathway, in TCA cycle and in ED pathway were 90% higher, 64% and 31% lower on Glc/Pro than on Glc, respectively. The maintenance energy on Glc/Pro was 58.4% lower than that on Glc. The energy charge was lower on Glc than on Glc/Pro, indicating that the cells on Glc suffered from energy burden. This study elucidates the impacts of proline on the central metabolism of S. erythraea and deepens the understanding of its metabolism.

  6. cis-Proline-mediated Ser(P)[superscript 5] Dephosphorylation by the RNA Polymerase II C-terminal Domain Phosphatase Ssu72

    Werner-Allen, Jon W.; Lee, Chul-Jin; Liu, Pengda; Nicely, Nathan I.; Wang, Su; Greenleaf, Arno L.; Zhou, Pei (Duke)


    RNA polymerase II coordinates co-transcriptional events by recruiting distinct sets of nuclear factors to specific stages of transcription via changes of phosphorylation patterns along its C-terminal domain (CTD). Although it has become increasingly clear that proline isomerization also helps regulate CTD-associated processes, the molecular basis of its role is unknown. Here, we report the structure of the Ser(P){sup 5} CTD phosphatase Ssu72 in complex with substrate, revealing a remarkable CTD conformation with the Ser(P){sup 5}-Pro{sup 6} motif in the cis configuration. We show that the cis-Ser(P){sup 5}-Pro{sup 6} isomer is the minor population in solution and that Ess1-catalyzed cis-trans-proline isomerization facilitates rapid dephosphorylation by Ssu72, providing an explanation for recently discovered in vivo connections between these enzymes and a revised model for CTD-mediated small nuclear RNA termination. This work presents the first structural evidence of a cis-proline-specific enzyme and an unexpected mechanism of isomer-based regulation of phosphorylation, with broad implications for CTD biology

  7. Delta1-pyrroline-5-carboxylic acid formed by proline dehydrogenase from the Bacillus subtilis ssp. natto expressed in Escherichia coli as a precursor for 2-acetyl-1-pyrroline.

    Huang, Tzou-Chi; Huang, Yi-Wen; Hung, Hui-Ju; Ho, Chi-Tang; Wu, Mei-Li


    Proline dehydrogenase (PRODH) catalyzes the biosynthesis of Delta1-pyrroline-5-carboxylic acid (P5C). The Bacillus subtilis subsp. natto gene for the proline dehydrogenase (BnPRODH) was cloned and expressed in Escherichia coli. Nucleotide sequence analysis of the clone revealed an open-reading frame that encodes 302 amino acid polypeptide with a calculated molecular mass of 34.5 kDa. The deduced amino acid sequence showed sequence similarity to bacterial PRODH and PutA of E. coli. The BnPRODH gene was cloned into pET21b and was expressed at a high level in E. coli BL21(DE3). The expressed protein was purified by using nickel ion affinity column chromatography to homogeneity before characterization. The purified recombinant BnPRODH was used to produce P5C. Model system composed of P5C and methylglyoxal was set up to study the formation of 2-acetyl-1-pyrroline. Our data showed that P5C, derived from the conversion of l-proline by the purified recombinant PRODH, might react directly with methylglyoxal to form 2-AP. P5C/methylglyoxal pathway represents the first report of a biological mechanism by which 2-AP may be synthesized in vitro by PRODH.

  8. Proline modulates the effect of bisphosphonate on calcium levels and adenosine triphosphate production in cell lines derived from bovine Echinococcus granulosus protoscoleces.

    Fuchs, A G; Echeverría, C I; Pérez Rojo, F G; Prieto González, E A; Roldán, E J A


    Bisphosphonates have been proposed as pharmacological agents against parasite and cancer cell growth. The effect of these compounds on helminthic cell viability and acellular compartment morphology, however, has not yet been studied. The effects of different types of bisphosphonates, namely etidronate (EHDP), pamidronate (APD), alendronate (ABP), ibandronate (IB) and olpadronate (OPD), and their interaction with amiloride, 1,25-dihydroxycholecalciferol (D3) and proline were evaluated on a cell line derived from bovine Echinococcus granulousus protoscoleces (EGPE) that forms cystic colonies in agarose. The EGPE cell line allowed testing the effect of bisphosphonates alone and in association with other compounds that could modulate calcium apposition/deposition, and were useful in measuring the impact of these compounds on cell growth, cystic colony formation and calcium storage. Decreased cell growth and cystic colony formation were found with EHDP, IB and OPD, and increased calcium storage with EHDP only. Calcium storage in EGPE cells appeared to be sensitive to the effect of amiloride, D3 and proline. Proline decreased calcium storage and increased colony formation. Changes in calcium storage may be associated with degenerative changes of the cysts, as shown in the in vitro colony model and linked to an adenosine triphosphate (ATP) decrease. In conclusion, bisphosphonates could be suitable tempering drugs to treat cestode infections.

  9. Structure and activity of the peptidyl-prolyl isomerase domain from the histone chaperone Fpr4 toward histone H3 proline isomerization.

    Monneau, Yoan R; Soufari, Heddy; Nelson, Christopher J; Mackereth, Cameron D


    The FK506-binding protein (FKBP) family of peptidyl-prolyl isomerases (PPIases) is characterized by a common catalytic domain that binds to the inhibitors FK506 and rapamycin. As one of four FKBPs within the yeast Saccharomyces cerevisiae, Fpr4 has been described as a histone chaperone, and is in addition implicated in epigenetic function in part due to its mediation of cis-trans conversion of proline residues within histone tails. To better understand the molecular details of this activity, we have determined the solution structure of the Fpr4 C-terminal PPIase domain by using NMR spectroscopy. This canonical FKBP domain actively increases the rate of isomerization of three decapeptides derived from the N terminus of yeast histone H3, whereas maintaining intrinsic cis and trans populations. Observation of the uncatalyzed and Fpr4-catalyzed isomerization rates at equilibrium demonstrate Pro(16) and Pro(30) of histone H3 as the major proline targets of Fpr4, with little activity shown against Pro(38). This alternate ranking of the three target prolines, as compared with affinity determination or the classical chymotrypsin-based fluorescent assay, reveals the mechanistic importance of substrate residues C-terminal to the peptidyl-prolyl bond.

  10. Core Transmembrane Domain 6 Plays a Pivotal Role in the Transport Cycle of the Sodium/Proline Symporter PutP.

    Bracher, Susanne; Schmidt, Claudia C; Dittmer, Sophie I; Jung, Heinrich


    Crystal structures of transporters with a LeuT-type structural fold assign core transmembrane domain 6 (TM6') a central role in substrate binding and translocation. Here, the function of TM6' in the sodium/proline symporter PutP, a member of the solute/sodium symporter family, was investigated. A complete scan of TM6' identified eight amino acids as particularly important for PutP function. Of these residues, Tyr-248, His-253, and Arg-257 impact sodium binding, whereas Arg-257 and Ala-260 may participate in interactions leading to closure of the inner gate. Furthermore, the previous suggestion of an involvement of Trp-244, Tyr-248, and Pro-252 in proline binding is further supported. In addition, substitution of Gly-245, Gly-247, and Gly-250 affects the amount of PutP in the membrane. A Cys accessibility analysis suggests an involvement of the inner half of TM6' in the formation of a hydrophilic pathway that is open to the inside in the absence of ligands and closed in the presence of sodium and proline. In conclusion, the results demonstrate that TM6' plays a central role in substrate binding and release on the inner side of the membrane also in PutP and extend the knowledge on functionally relevant amino acids in transporters with a LeuT-type structural fold.

  11. Regulatory region with putA gene of proline dehydrogenase that links to the lum and the lux operons in Photobacterium leiognathi.

    Lin, J W; Yu, K Y; Chen, H Y; Weng, S F


    Nucleotide sequence of regulatory region (R & R) with putA gene (EMBL Accession No. U39227) from Photobacterium leiognathi PL741 has been determined, and the putA gene encoded amino acid sequence of proline dehydrogenase is deduced. Alignment and comparison of proline dehydrogenase of P. leiognathi with the proline dehydrogenase domain in the PutA protein of Escherichia coli and Salmonella typhimurium show that they are homologous. Nucleotide sequence reveals that regulatory region with the putA gene is linked to the lum and lux operons in genome; the gene order is (R & R: regulatory region; ter:transcriptional terminator), whereas the R & R is the regulatory region for the lum and the lux operons, ter is the transcriptional terminator for the lum operon, and R & R(I) apparently is the regulatory region for the putA and related genes. Nucleotide sequence analysis illustrates the specific inverted repeat (SIR), cAMP-CRP consensus sequence, canonical -10/-35 promoter, putative operator and Shine-Dalgarno (SD) sequence on the regulatory region R & R(I) for the putA and related genes; it suggests that the putA and related genes are simply linked to the lum and the lux operons in genome, the regulatory region R & R(I) is independent for the putA and related genes.

  12. Reversions of two proline-requiring auxotrophs of Haemophilus influenzae by N-methyl-N'-nitro-N-nitrosoguanidine and hydrazine

    Kimball, R.F.


    New mutation detection systems are described for Haemophilus influenzae. They involve two independently isolated proline auxotrophs which appear to be mutants at different sites in a proline locus (proB) that is very closely linked to a locus (thd) for thymidine requirement. One of the mutants, proB1, appears to revert to prototrophy only by mutations at the locus. The other, proB2, reverts both by mutation at the locus and by unlinked suppressors. The latter account for about 90 percent of the reversions induced by MNNG and by HZ. The close linkage of proB to thd was used to distinguish between true revertants and suppressors by a transformation test. A comparison was made between the mutation induction kinetics of the different classes of revertants and mutations to novobiocin resistance with MNNG and HZ. The very different induction kinetics for these two mutagens previously reported for the novobiocin resistance system were also found for the proline systems. There were some differences between the detection systems, however, in the frequency of induced mutation relative to the spontaneous frequency and, in one case, in the form of the induction curve. It is concluded that the major features of the induction curves reflect the amount of damage done to DNA and so are general for all systems, but that there are some features which are locus- or site-specific.

  13. Prey-induced changes in the accumulation of amino acids and phenolic metabolites in the leaves of Drosera capensis L.

    Kováčik, Jozef; Klejdus, Bořivoj; Stork, František; Hedbavny, Josef


    Effect of prey feeding (ants Formica fusca) on the quantitative changes in the accumulation of free amino acids, soluble proteins, phenolic metabolites and mineral nutrients in the leaves of carnivorous plant Drosera capensis was studied. Arginine was the most abundant compound in Drosera leaves, while proline was abundant in ants. The amount of the majority of amino acids and their sum were elevated in the fed leaves after 3 and 21 days, and the same, but with further enhancement after 21 days, was observed in ants. Accumulation of amino acids also increased in young non-fed leaves of fed plants. Soluble proteins decreased in ants, but were not enhanced in fed leaves. This confirms the effectiveness of sundew's enzymatic machinery in digestion of prey and suggests that amino acids are not in situ deposited, but rather are allocated within the plant. The content of total soluble phenols, flavonoids and two selected flavonols (quercetin and kaempferol) was not affected by feeding in Drosera leaves, indicating that their high basal level was sufficient for the plant's metabolism and prey-induced changes were mainly N based. The prey also showed to be an important source of other nutrients besides N, and a stimulation of root uptake of some mineral nutrients is assumed (Mg, Cu, Zn). Accumulation of Ca and Na was not affected by feeding.

  14. PABLM. Accumulated Environment Radiation Dose

    Napier, B.A.; Kennedy, W.E.Jr.; Soldat, J.K. [Pacific Northwest Lab., Richland, WA (United States)


    PABLM calculates internal radiation doses to man from radionuclides in food products and external radiation doses from radionuclides in the environment. Radiation doses from radionuclides in the environment may be calculated from deposition on the soil or plants during an atmospheric or liquid release, or from exposure to residual radionuclides after the releases have ended. Radioactive decay is considered during the release, after deposition, and during holdup of food after harvest. The radiation dose models consider exposure to radionuclides deposited on the ground or crops from contaminated air or irrigation water, radionuclides in contaminated drinking water, aquatic foods raised in contaminated water, and radionuclides in bodies of water and sediments where people might fish, boat, or swim. For vegetation, the radiation dose model considers both direct deposition and uptake through roots. Doses may be calculated for either a maximum-exposed individual or for a population group. The program is designed to calculate accumulated radiation doses from the chronic ingestion of food products that contain radionuclides and doses from the external exposure to radionuclides in the environment. A first-year committed dose is calculated as well as an integrated dose for a selected number of years.

  15. Sequential biases in accumulating evidence

    Huggins, Richard; Dogo, Samson Henry


    Whilst it is common in clinical trials to use the results of tests at one phase to decide whether to continue to the next phase and to subsequently design the next phase, we show that this can lead to biased results in evidence synthesis. Two new kinds of bias associated with accumulating evidence, termed ‘sequential decision bias’ and ‘sequential design bias’, are identified. Both kinds of bias are the result of making decisions on the usefulness of a new study, or its design, based on the previous studies. Sequential decision bias is determined by the correlation between the value of the current estimated effect and the probability of conducting an additional study. Sequential design bias arises from using the estimated value instead of the clinically relevant value of an effect in sample size calculations. We considered both the fixed‐effect and the random‐effects models of meta‐analysis and demonstrated analytically and by simulations that in both settings the problems due to sequential biases are apparent. According to our simulations, the sequential biases increase with increased heterogeneity. Minimisation of sequential biases arises as a new and important research area necessary for successful evidence‐based approaches to the development of science. © 2015 The Authors. Research Synthesis Methods Published by John Wiley & Sons Ltd. PMID:26626562

  16. Quantum gravity and inventory accumulation

    Sheffield, Scott


    We begin by studying inventory accumulation at a LIFO (last-in-first-out) retailer with two products. In the simplest version, the following occur with equal probability at each time step: first product ordered, first product produced, second product ordered, second product produced. The inventory thus evolves as a simple random walk on Z^2. In more interesting versions, a p fraction of customers orders the "freshest available" product regardless of type. We show that the corresponding random walks scale to Brownian motions with diffusion matrices depending on p. We then turn our attention to the critical Fortuin-Kastelyn random planar map model, which gives, for each q>0, a probability measure on random (discretized) two-dimensional surfaces decorated by loops, related to the q-state Potts model. A longstanding open problem is to show that as the discretization gets finer, the surfaces converge in law to a limiting (loop-decorated) random surface. The limit is expected to be a Liouville quantum gravity surfa...

  17. Towards understanding the tandem mass spectra of protonated oligopeptides. 2: The proline effect in collision-induced dissociation of protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp).

    Bleiholder, Christian; Suhai, Sándor; Harrison, Alex G; Paizs, Béla


    The product ion spectra of proline-containing peptides are commonly dominated by y(n) ions generated by cleavage at the N-terminal side of proline residues. This proline effect is investigated in the current work by collision-induced dissociation (CID) of protonated Ala-Ala-Xxx-Pro-Ala (Xxx includes Ala, Ser, Leu, Val, Phe, and Trp) in an electrospray/quadrupole/time-of-flight (QqTOF) mass spectrometer and by quantum chemical calculations on protonated Ala-Ala-Ala-Pro-Ala. The CID spectra of all investigated peptides show a dominant y(2) ion (Pro-Ala sequence). Our computational results show that the proline effect mainly arises from the particularly low threshold energy for the amide bond cleavage N-terminal to the proline residue, and from the high proton affinity of the proline-containing C-terminal fragment produced by this cleavage. These theoretical results are qualitatively supported by the experimentally observed y(2)/b(3) abundance ratios for protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp). In the post-cleavage phase of fragmentation the N-terminal oxazolone fragment with the Ala-Ala-Xxx sequence and Pro-Ala compete for the ionizing proton for these peptides. As the proton affinity of the oxazolone fragment increases, the y(2)/b(3) abundance ratio decreases.

  18. Towards Understanding the Tandem Mass Spectra of Protonated Oligopeptides. 2: The Proline Effect in Collision-Induced Dissociation of Protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp)

    Bleiholder, Christian; Suhai, Sándor; Harrison, Alex G.; Paizs, Béla


    The product ion spectra of proline-containing peptides are commonly dominated by y n ions generated by cleavage at the N-terminal side of proline residues. This proline effect is investigated in the current work by collision-induced dissociation (CID) of protonated Ala-Ala-Xxx-Pro-Ala (Xxx includes Ala, Ser, Leu, Val, Phe, and Trp) in an electrospray/quadrupole/time-of-flight (QqTOF) mass spectrometer and by quantum chemical calculations on protonated Ala-Ala-Ala-Pro-Ala. The CID spectra of all investigated peptides show a dominant y 2 ion (Pro-Ala sequence). Our computational results show that the proline effect mainly arises from the particularly low threshold energy for the amide bond cleavage N-terminal to the proline residue, and from the high proton affinity of the proline-containing C-terminal fragment produced by this cleavage. These theoretical results are qualitatively supported by the experimentally observed y 2 / b 3 abundance ratios for protonated Ala-Ala-Xxx-Pro-Ala (Xxx = Ala, Ser, Leu, Val, Phe, and Trp). In the post-cleavage phase of fragmentation the N-terminal oxazolone fragment with the Ala-Ala-Xxx sequence and Pro-Ala compete for the ionizing proton for these peptides. As the proton affinity of the oxazolone fragment increases, the y 2 / b 3 abundance ratio decreases.

  19. Accumulation boundaries: codimension-two accumulation of accumulations in phase diagrams of semiconductor lasers, electric circuits, atmospheric and chemical oscillators.

    Bonatto, Cristian; Gallas, Jason Alfredo Carlson


    We report high-resolution phase diagrams for several familiar dynamical systems described by sets of ordinary differential equations: semiconductor lasers; electric circuits; Lorenz-84 low-order atmospheric circulation model; and Rössler and chemical oscillators. All these systems contain chaotic phases with highly complicated and interesting accumulation boundaries, curves where networks of stable islands of regular oscillations with ever-increasing periodicities accumulate systematically. The experimental exploration of such codimension-two boundaries characterized by the presence of infinite accumulation of accumulations is feasible with existing technology for some of these systems.

  20. Erythrocytosis-associated HIF-2α Mutations Demonstrate a Critical Role for Residues C-terminal to the Hydroxylacceptor Proline*

    Furlow, Paul W.; Percy, Melanie J.; Sutherland, Scott; Bierl, Charlene; McMullin, Mary Frances; Master, Stephen R.; Lappin, Terence R. J.; Lee, Frank S.


    A classic physiologic response to hypoxia in humans is the up-regulation of the ERYTHROPOIETIN (EPO) gene, which is the central regulator of red blood cell mass. The EPO gene, in turn, is activated by hypoxia inducible factor (HIF). HIF is a transcription factor consisting of an α subunit (HIF-α) and a β subunit (HIF-β). Under normoxic conditions, prolyl hydroxylase domain protein (PHD, also known as HIF prolyl hydroxylase and egg laying-defective nine protein) site specifically hydroxylates HIF-α in a conserved LXXLAP motif (where underlining indicates the hydroxylacceptor proline). This provides a recognition motif for the von Hippel Lindau protein, a component of an E3 ubiquitin ligase complex that targets hydroxylated HIF-α for degradation. Under hypoxic conditions, this inherently oxygen-dependent modification is arrested, thereby stabilizing HIF-α and allowing it to activate the EPO gene. We previously identified and characterized an erythrocytosis-associated HIF2A mutation, G537W. More recently, we reported two additional erythrocytosis-associated HIF2A mutations, G537R and M535V. Here, we describe the functional characterization of these two mutants as well as a third novel erythrocytosis-associated mutation, P534L. These mutations affect residues C-terminal to the LXXLAP motif. We find that all result in impaired degradation and thus aberrant stabilization of HIF-2α. However, each exhibits a distinct profile with respect to their effects on PHD2 binding and von Hippel Lindau interaction. These findings reinforce the importance of HIF-2α in human EPO regulation, demonstrate heterogeneity of functional defects arising from these mutations, and point to a critical role for residues C-terminal to the LXXLAP motif in HIF-α. PMID:19208626