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Sample records for aggrecan c-type lectin

  1. Tetranectin, a trimeric plasminogen-binding C-type lectin

    DEFF Research Database (Denmark)

    Holtet, T L; Graversen, Jonas Heilskov; Clemmensen, I;

    1997-01-01

    Tetranectin, a plasminogen-binding protein belonging to the family of C-type lectins, was expressed in E. coli and converted to its native form by in vitro refolding and proteolytic processing. Recombinant tetranectin-as well as natural tetranectin from human plasma-was shown by chemical cross......-linking analysis and SDS-PAGE to be a homo-trimer in solution as are other known members of the collectin family of C-type lectins. Biochemical evidence is presented showing that an N-terminal domain encoded within exons 1 and 2 of the tetranectin gene is necessary and sufficient to govern subunit trimerization....

  2. C-type lectins%C型凝集素

    Institute of Scientific and Technical Information of China (English)

    谢建辉; 顾建新

    2011-01-01

    C型凝集素(C-type lectin)代表一个识别碳水化合物配体依赖于钙离子(Ca2+)参与的糖原结合蛋白家族,含有一个或多个一级结构和二级结构同源的碳水化合物识别结构域.随着研究的深入,越来越多的C型凝集素能够识别体内的非糖类的配体,包括蛋白质和脂类等.这些C型凝集素在维持机体稳态、免疫防御以及免疫监视等重要生理病理过程中发挥着重要作用.就C型凝集素的结构、分类和在免疫系统中的功能作一介绍.%C-type lectins are Ca2+-dependent glycan-binding proteins and share primary and secondary structural homology in their carbohydrate-recognition domains (CRDs). However, many members of this family are recently identified not to bind carbohydrates and have evolved to recognize non-sugar ligands such as proteins and lipids. The large family of C-type lectins has an important role in the physiological functions and pathological processes including immune homeostasis, immune defenses, and immune surveillance and so on. In this short review, we summarize the structure of C-type lectin domain, the classification of C-type lectins and their role in the immune system.

  3. DMPD: C-type lectin receptors in antifungal immunity. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 18160296 C-type lectin receptors in antifungal immunity. Willment JA, Brown GD. Tre...nds Microbiol. 2008 Jan;16(1):27-32. Epub 2007 Dec 21. (.png) (.svg) (.html) (.csml) Show C-type lectin receptors in antifungal... immunity. PubmedID 18160296 Title C-type lectin receptors in antifungal immunity. Author

  4. C-type lectins do not act as functional receptors for filovirus entry into cells

    Energy Technology Data Exchange (ETDEWEB)

    Matsuno, Keita; Nakayama, Eri; Noyori, Osamu [Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Sapporo (Japan); Marzi, Andrea; Ebihara, Hideki [Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, MT (United States); Irimura, Tatsuro [Graduate School of Pharmaceutical Science, University of Tokyo, Tokyo (Japan); Feldmann, Heinz [Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, Hamilton, MT (United States); Takada, Ayato, E-mail: atakada@czc.hokudai.ac.jp [Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Sapporo (Japan)

    2010-12-03

    Research highlights: {yields} Filovirus glycoprotein (GP) having a deficient receptor binding region were generated. {yields} Mutant GPs mediated virus entry less efficiently than wild-type GP. {yields} Mutant GPs bound to C-type lectins but not mediated entire steps of cellular entry. {yields} C-type lectins do not independently mediate filovirus entry into cells. {yields} Other molecule(s) are required for C-type lectin-mediated entry of filoviruses. -- Abstract: Cellular C-type lectins have been reported to facilitate filovirus infection by binding to glycans on filovirus glycoprotein (GP). However, it is not clearly known whether interaction between C-type lectins and GP mediates all the steps of virus entry (i.e., attachment, internalization, and membrane fusion). In this study, we generated vesicular stomatitis viruses pseudotyped with mutant GPs that have impaired structures of the putative receptor binding regions and thus reduced ability to infect the monkey kidney cells that are routinely used for virus propagation. We found that infectivities of viruses with the mutant GPs dropped in C-type lectin-expressing cells, parallel with those in the monkey kidney cells, whereas binding activities of these GPs to the C-type lectins were not correlated with the reduced infectivities. These results suggest that C-type lectin-mediated entry of filoviruses requires other cellular molecule(s) that may be involved in virion internalization or membrane fusion.

  5. C-type lectins do not act as functional receptors for filovirus entry into cells

    International Nuclear Information System (INIS)

    Research highlights: → Filovirus glycoprotein (GP) having a deficient receptor binding region were generated. → Mutant GPs mediated virus entry less efficiently than wild-type GP. → Mutant GPs bound to C-type lectins but not mediated entire steps of cellular entry. → C-type lectins do not independently mediate filovirus entry into cells. → Other molecule(s) are required for C-type lectin-mediated entry of filoviruses. -- Abstract: Cellular C-type lectins have been reported to facilitate filovirus infection by binding to glycans on filovirus glycoprotein (GP). However, it is not clearly known whether interaction between C-type lectins and GP mediates all the steps of virus entry (i.e., attachment, internalization, and membrane fusion). In this study, we generated vesicular stomatitis viruses pseudotyped with mutant GPs that have impaired structures of the putative receptor binding regions and thus reduced ability to infect the monkey kidney cells that are routinely used for virus propagation. We found that infectivities of viruses with the mutant GPs dropped in C-type lectin-expressing cells, parallel with those in the monkey kidney cells, whereas binding activities of these GPs to the C-type lectins were not correlated with the reduced infectivities. These results suggest that C-type lectin-mediated entry of filoviruses requires other cellular molecule(s) that may be involved in virion internalization or membrane fusion.

  6. C-type Lectin Receptors for Tumor Eradication: Future Directions

    Energy Technology Data Exchange (ETDEWEB)

    Streng-Ouwehand, Ingeborg; Unger, Wendy W. J.; Kooyk, Yvette van, E-mail: y.vankooyk@vumc.nl [Department of Molecular Cell Biology and Immunology, VU University Medical Center, P.O. Box 7057, 1007 MB Amsterdam (Netherlands)

    2011-08-08

    Dendritic cells are key regulators in directing immune responses and therefore are under extensive research for the induction of anti-tumor responses. DCs express a large array of receptors by which they scan their surroundings for recognition and uptake of pathogens. One of the receptor-families is the C-type lectins (CLR), which bind carbohydrate structures and internalize antigens upon recognition. Intracellular routing of antigen through CLR enhances loading and presentation of antigen through MHC class I and II, inducing antigen-specific CD4{sup +} and CD8{sup +} T-cell proliferation and skewing T-helper cells. These characteristics make CLRs very interesting targets for DC-based immunotherapy. Profound research has been done on targeting specific tumor antigens to CLR using either antibodies or the natural ligands such as glycan structures. In this review we will focus on the current data showing the potency of CLR-targeting and discuss improvements that can be achieved to enhance anti-tumor activity in the near future.

  7. Mosquito C-type lectins maintain gut microbiome homeostasis.

    Science.gov (United States)

    Pang, Xiaojing; Xiao, Xiaoping; Liu, Yang; Zhang, Rudian; Liu, Jianying; Liu, Qiyong; Wang, Penghua; Cheng, Gong

    2016-01-01

    The long-term evolutionary interaction between the host immune system and symbiotic bacteria determines their cooperative rather than antagonistic relationship. It is known that commensal bacteria have evolved a number of mechanisms to manipulate the mammalian host immune system and maintain homeostasis. However, the strategies employed by the microbiome to overcome host immune responses in invertebrates still remain to be understood. Here, we report that the gut microbiome in mosquitoes utilizes C-type lectins (mosGCTLs) to evade the bactericidal capacity of antimicrobial peptides (AMPs). Aedes aegypti mosGCTLs facilitate colonization by multiple bacterial strains. Furthermore, maintenance of the gut microbial flora relies on the expression of mosGCTLs in A. aegypti. Silencing the orthologues of mosGCTL in another major mosquito vector (Culex pipiens pallens) also impairs the survival of gut commensal bacteria. The gut microbiome stimulates the expression of mosGCTLs, which coat the bacterial surface and counteract AMP activity. Our study describes a mechanism by which the insect symbiotic microbiome offsets gut immunity to achieve homeostasis. PMID:27572642

  8. Dual function of C-type lectin-like receptors in the immune system.

    NARCIS (Netherlands)

    Cambi, A.; Figdor, C.G.

    2003-01-01

    Carbohydrate-binding C-type lectin and lectin-like receptors play an important role in the immune system. The large family can be subdivided into subtypes according to their structural similarities and functional differences. The selectins are of major importance in mediating cell adhesion and migra

  9. Chicken lung lectin is a functional C-type lectin and inhibits haemagglutination by influenza A virus.

    NARCIS (Netherlands)

    Hogenkamp, A.; Isohadouten, N.; Reemers, S.S.N.; Romijn, R.A.; Hemrika, W.; White, M.R.; Tefsen, B.; Vervelde, L.; van Eijk, M.; Veldhuizen, E.J.A.; Haagsman, H.P.

    2008-01-01

    Many proteins of the calcium-dependent (C-type) lectin family have been shown to play an important role in innate immunity. They can bind to a broad range of carbohydrates, which enables them to interact with ligands present on the surface of micro-organisms.We previously reported the finding of a n

  10. Antimicrobial properties of avian eggshell-specific C-type lectin-like proteins.

    Science.gov (United States)

    Wellman-Labadie, Olivier; Lakshminarayanan, Rajamani; Hincke, Maxwell T

    2008-03-01

    C-type lectin-like proteins are major components of the calcified eggshell of multiple avian species. In this study, two representative avian C-type lectin-like proteins, ovocleidin-17 and ansocalcin, were purified from decalcified chicken and goose eggshell protein extracts and investigated for carbohydrate binding activity as well as antimicrobial activity. Purified ovocleidin-17 and ansocalcin were found to bind bacterial polysaccharides, and were bactericidal against Bacillus subtilis, Staphylococcus aureus and Pseudomona aeruginosa. Bactericidal activity was found to be enhanced in the presence of calcium but was not dependent on its presence. The results suggest that avian C-type lectin-like proteins may play an important antimicrobial role in defence of the avian embryo. PMID:18258195

  11. A C-type lectin from Bothrops jararacussu venom disrupts Staphylococcal biofilms.

    Directory of Open Access Journals (Sweden)

    Raphael Contelli Klein

    Full Text Available Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chromatography, and the fractions were tested against S. aureus. Biofilm growth, but not bacterial growth, was affected by several fractions. Two fractions (15 and 16 showed the best activities and were also assayed against S. epidermidis NRS101. Fraction 15 was identified by TripleTOF mass spectrometry as a galactose-binding C-type lectin with a molecular weight of 15 kDa. The lectin was purified from the crude venom by D-galactose affinity chromatography, and only one peak was observed. This pure lectin was able to inhibit 75% and 80% of S. aureus and S. epidermidis biofilms, respectively, without affecting bacterial cell viability. The lectin also exhibited a dose-dependent inhibitory effect on both bacterial biofilms. The antibiofilm activity was confirmed using scanning electron microscopy. A pre-formed S. epidermidis biofilm was significantly disrupted by the C-type lectin in a time-dependent manner. Additionally, the lectin demonstrated the ability to inhibit biofilm formation by several mastitis pathogens, including different field strains of S. aureus, S. hyicus, S. chromogenes, Streptococcus agalactiae, and Escherichia coli. These findings reveal a new activity for C-type lectins. Studies are underway to evaluate the biological activity of these lectins in a mouse mastitis model.

  12. Purification and biological effects of a C-type lectin isolated from Bothrops moojeni

    Directory of Open Access Journals (Sweden)

    PSF Barbosa

    2010-01-01

    Full Text Available Snake venom proteins from the C-type lectin family have very distinct biological activities despite their highly conserved primary structure, which is homologous to the carbohydrate recognition region of true C-type lectins. We purified a lectin-like protein (BmLec from Bothrops moojeni venom and investigated its effect on platelet aggregation, insulin secretion, antibacterial activity, and isolated kidney cells. The BmLec was purified using two chromatographic steps: affinity chromatography and reverse phase high performance liquid chromatography (HPLC. BmLec showed a dose-dependent platelet aggregation and significantly decreased the bacterial growth rate in approximately 15%. During scanning electron microscopy, the profile of Xanthomonas axonopodis pv. passiflorae treated with lectin disclosed a high vesiculation and membrane rupture. BmLec induced a strong and significant increase in insulin secretion at 2.8 and 16.7 mM glucose concentrations, and this effect was seen in the presence of EGTA in both experiments. BmLec (10 µg/mL increased the perfusion pressure, renal vascular resistance and urinary flow. The glomerular filtration rate and percentages of sodium, potassium and chloride tubular transport were reduced at 60 minutes of perfusion. Renal alterations caused by BmLec were completely inhibited by indomethacin in all evaluated parameters. In conclusion, the C-type lectin isolated from Bothrops moojeni affected platelet aggregation, insulin secretion, antibacterial activity and isolated kidney function.

  13. Structure of the C-type lectin carbohydrate recognition domain of human tetranectin

    DEFF Research Database (Denmark)

    Kastrup, J S; Nielsen, B B; Rasmussen, H;

    1998-01-01

    Tetranectin (TN) is a C-type lectin involved in fibrinolysis, being the only endogenous ligand known to bind specifically to the kringle 4 domain of plasminogen. TN was originally isolated from plasma, but shows a wide tissue distribution. Furthermore, TN has been found in the extracellular matrix...

  14. Targeting of macrophage galactose-type C-type lectin (MGL) induces DC signaling and activation

    DEFF Research Database (Denmark)

    Napoletano, Chiara; Zizzari, Ilaria G; Rughetti, Aurelia;

    2012-01-01

    Dendritic cells (DCs) sense the microenvironment through several types of receptors recognizing pathogen-associated molecular patterns. In particular, C-type lectins, expressed by distinct subsets of DCs, recognize and internalize specific carbohydrate antigen in a Ca(2+) -dependent manner. Targe...

  15. Transmission-blocking antibodies against mosquito C-type lectins for dengue prevention.

    Science.gov (United States)

    Liu, Yang; Zhang, Fuchun; Liu, Jianying; Xiao, Xiaoping; Zhang, Siyin; Qin, Chengfeng; Xiang, Ye; Wang, Penghua; Cheng, Gong

    2014-02-01

    C-type lectins are a family of proteins with carbohydrate-binding activity. Several C-type lectins in mammals or arthropods are employed as receptors or attachment factors to facilitate flavivirus invasion. We previously identified a C-type lectin in Aedes aegypti, designated as mosquito galactose specific C-type lectin-1 (mosGCTL-1), facilitating the attachment of West Nile virus (WNV) on the cell membrane. Here, we first identified that 9 A. aegypti mosGCTL genes were key susceptibility factors facilitating DENV-2 infection, of which mosGCTL-3 exhibited the most significant effect. We found that mosGCTL-3 was induced in mosquito tissues with DENV-2 infection, and that the protein interacted with DENV-2 surface envelop (E) protein and virions in vitro and in vivo. In addition, the other identified mosGCTLs interacted with the DENV-2 E protein, indicating that DENV may employ multiple mosGCTLs as ligands to promote the infection of vectors. The vectorial susceptibility factors that facilitate pathogen invasion may potentially be explored as a target to disrupt the acquisition of microbes from the vertebrate host. Indeed, membrane blood feeding of antisera against mosGCTLs dramatically reduced mosquito infective ratio. Hence, the immunization against mosGCTLs is a feasible approach for preventing dengue infection. Our study provides a future avenue for developing a transmission-blocking vaccine that interrupts the life cycle of dengue virus and reduces disease burden. PMID:24550728

  16. Transmission-blocking antibodies against mosquito C-type lectins for dengue prevention.

    Directory of Open Access Journals (Sweden)

    Yang Liu

    2014-02-01

    Full Text Available C-type lectins are a family of proteins with carbohydrate-binding activity. Several C-type lectins in mammals or arthropods are employed as receptors or attachment factors to facilitate flavivirus invasion. We previously identified a C-type lectin in Aedes aegypti, designated as mosquito galactose specific C-type lectin-1 (mosGCTL-1, facilitating the attachment of West Nile virus (WNV on the cell membrane. Here, we first identified that 9 A. aegypti mosGCTL genes were key susceptibility factors facilitating DENV-2 infection, of which mosGCTL-3 exhibited the most significant effect. We found that mosGCTL-3 was induced in mosquito tissues with DENV-2 infection, and that the protein interacted with DENV-2 surface envelop (E protein and virions in vitro and in vivo. In addition, the other identified mosGCTLs interacted with the DENV-2 E protein, indicating that DENV may employ multiple mosGCTLs as ligands to promote the infection of vectors. The vectorial susceptibility factors that facilitate pathogen invasion may potentially be explored as a target to disrupt the acquisition of microbes from the vertebrate host. Indeed, membrane blood feeding of antisera against mosGCTLs dramatically reduced mosquito infective ratio. Hence, the immunization against mosGCTLs is a feasible approach for preventing dengue infection. Our study provides a future avenue for developing a transmission-blocking vaccine that interrupts the life cycle of dengue virus and reduces disease burden.

  17. Quantitative Expression of C-Type Lectin Receptors in Humans and Mice

    OpenAIRE

    Hans-Joachim Anders; Roman Günthner; Regina Gröbmayr; Christoph Römmele; Heni Eka Susanti; Maciej Lech

    2012-01-01

    C-type lectin receptors, their adaptor molecules and S-type lectins (galectins) are involved in the recognition of glycosylated self-antigens and pathogens. However, little is known about the species- and organ-specific expression profiles of these molecules. We therefore determined the mRNA expression levels of Dectin-1, MR1, MR2, DC-SIGN, Syk, Card-9, Bcl-10, Malt-1, Src, Dec-205, Galectin-1, Tim-3, Trem-1, and DAP-12 in 11 solid organs of human and mice. Mouse organs revealed lower mRNA...

  18. The role of Syk/CARD9 coupled C-type lectins in antifungal immunity

    OpenAIRE

    Drummond, Rebecca A.; Saijo, Shinobu; Iwakura, Yoichiro; Brown, Gordon D.

    2010-01-01

    Fungal infections are affecting an increasing number of people, and the failure of current therapies in treating systemic infection has resulted in an unacceptably high mortality rate. It is therefore of importance that we understand immune mechanisms operating during fungal infections, in order to facilitate development of adjunctive immunotherapies for the treatment of these diseases. C-type lectin receptors (CLRs) are pattern recognition receptors (PRRs) that are critical for immune respon...

  19. Analysis of C-type lectin receptor induced NF-kappaB signaling

    OpenAIRE

    Straßer, Andreas Dominikus

    2014-01-01

    Myeloid C-type lectin receptors (CLRs) that signal via Syk and the central Card9-Bcl10-Malt1 (CBM) complex induce the transcription of NF-κB-regulated genes. Activation of those receptors mediates inflammatory reactions and the defense against various pathogens. Despite the non-redundant role of CLRs for the induction of innate immune responses, particularly receptor-proximal events that transduce ligand binding to downstream signaling remain to be defined. This dissertation identifies PKCδ a...

  20. Differential expression of two C-type lectins in grass carp Ctenopharyngodon idella and their response to grass carp reovirus.

    Science.gov (United States)

    Ju, C S; He, L B; Pei, Y Y; Jiang, Y; Huang, R; Li, Y M; Liao, L J; Jang, S H; Wang, Y P

    2016-02-01

    The cDNAs of two C-type lectins in grass carp Ctenopharyngodon idella, galactose-binding lectin (galbl) and mannose-binding lectin (mbl), were cloned and analysed in this study. Both of them exhibited the highest expression level in liver, whereas their expression pattern differed in early phase of embryonic development. Following exposure to grass carp reovirus (GCRV), the mRNA expression level of galbl and mbl was significantly up-regulated in liver and intestine. PMID:26643267

  1. Molecular cloning and characterization of a C-type lectin from Ancylostoma ceylanicum: evidence for a role in hookworm reproductive physiology.

    OpenAIRE

    Brown, Allison C.; Harrison, Lisa M.; Kapulkin, Wadim; Jones, Brian F.; Sinha, Anindita; Savage, Amy; Villalon, Nicholas; Cappello, Michael

    2006-01-01

    Lectins comprise a family of related proteins that mediate essential cell functions through binding to carbohydrates. Within this protein family, C-type lectins are defined by the requirement of calcium for optimal biologic activity. Using reverse transcription PCR, a cDNA corresponding to a putative C-type lectin has been amplified from the hookworm parasite Ancylostoma ceylanicum. The 550 nucleotide open reading frame of the Ancylostoma ceylanicum C-type Lectin-1 (AceCTL-1) cDNA corresponds...

  2. Sensing of cell death by myeloid C-type lectin receptors

    Science.gov (United States)

    Sancho, David; Reis e Sousa, Caetano

    2015-01-01

    Molecules associated with dead or dying cells can be detected by receptors on macrophages and dendritic cells. Signals from these receptors impact myeloid cell function and play a role in determining whether death is silent or proinflammatory, tolerogenic or immunogenic. Prominent among myeloid receptors detecting dead cells are C-type lectin receptors (CLRs). Signals from these receptors variably induce endocytosis of cell corpses, corpse degradation, retrieval of dead cell-associated antigens and/or modulation of immune responses. The sensing of tissue damage by myeloid CLRs complements detection of pathogens in immunity and represents an ancient response aimed at restoring tissue homeostasis. PMID:23332826

  3. Intestinally secreted C-type lectin Reg3b attenuates salmonellosis but not listeriosis in mice

    OpenAIRE

    Ampting, Van, M.T.J.; Loonen, L.M.P.; Schonewille, A.J.; Konings, I.; Vink, C; Iovanna, J.; Chamaillard, M; Dekker, J.; Meer, Van Der; Wells, J.; Bovee-Oudenhoven, I.M.J.

    2012-01-01

    The Reg3 protein family, including the human member designated pancreatitis-associated protein (PAP), consists of secreted proteins that contain a C-type lectin domain involved in carbohydrate binding. They are expressed by intestinal epithelial cells. Colonization of germ-free mice and intestinal infection with pathogens increase the expression of Reg3g and Reg3b in the murine ileum. Reg3g is directly bactericidal for Gram-positive bacteria, but the exact role of Reg3b in bacterial infection...

  4. Galactose recognition by a tetrameric C-type lectin, CEL-IV, containing the EPN carbohydrate recognition motif.

    OpenAIRE

    Hatakeyama, Tomomitsu; Kamiya, Takuro; Kusunoki, Masami; Nakamura-Tsuruta, Sachiko; Hirabayashi, Jun; Goda, Shuichiro; Unno, Hideaki

    2011-01-01

    CEL-IV is a C-type lectin isolated from a sea cucumber, Cucumaria echinata. This lectin is composed of four identical C-type carbohydrate-recognition domains (CRDs). X-ray crystallographic analysis of CEL-IV revealed that its tetrameric structure was stabilized by multiple interchain disulfide bonds among the subunits. Although CEL-IV has the EPN motif in its carbohydrate-binding sites, which is known to be characteristic of mannose binding C-type CRDs, it showed preferential binding of galac...

  5. Galactose Recognition by a Tetrameric C-type Lectin, CEL-IV, Containing the EPN Carbohydrate Recognition Motif*

    OpenAIRE

    Hatakeyama, Tomomitsu; Kamiya, Takuro; Kusunoki, Masami; Nakamura-Tsuruta, Sachiko; Hirabayashi, Jun; Goda, Shuichiro; Unno, Hideaki

    2011-01-01

    CEL-IV is a C-type lectin isolated from a sea cucumber, Cucumaria echinata. This lectin is composed of four identical C-type carbohydrate-recognition domains (CRDs). X-ray crystallographic analysis of CEL-IV revealed that its tetrameric structure was stabilized by multiple interchain disulfide bonds among the subunits. Although CEL-IV has the EPN motif in its carbohydrate-binding sites, which is known to be characteristic of mannose binding C-type CRDs, it showed preferential binding of galac...

  6. Molecular characterization and expression analysis of a novel dual-CRD C-type lectin in kuruma shrimp (Marsupenaeus japonicus)

    Institute of Scientific and Technical Information of China (English)

    ZHANG Man; MAO Yong; WANG Jun; FENG Wenrong; SONG Xiaohong; SU Yongquan

    2015-01-01

    C-type lectins are among the most significant pattern recognition receptors (PRRs) found in invertebrate. They are a class of carbohydrate-binding proteins that can recognize specific sugar moieties on the surface of pathogens. In the present study, a novel C-type lecitn (termed MjLectin) from kuruma shrimp Marsupenaeus japonicus was identified. The full-length cDNA of MjLectin was 1 245 bp with a 1 011 bp open reading frame (ORF) that encoded a polypeptide of 336 amino acid residues. MjLectin consisted of two tandemly arrayed carbohydrate-recognition domains (CRDs), unlike other reported M. japonicus C-type lectins with only one CRD. It showed a high similarity to other shrimp dual-CRD lectins. Among the Ca2+-binding Site 2, the tripeptide motif dictating the carbohydrate binding specificity was exhibited as a rare mutant LPN (Leu134-Pro135-Asn136) in CRD1 and a traditional EPN (Glu299-Pro300-Asn301) in CRD2, respectively. MjLectin showed a specific expression pattern in both tissue and cellular levels, for its mRNA transcript was mainly expressed in the F-cells of the hepatopancreas. After white spot syndrome virus (WSSV) challenge (3.6×108 virions/μL), the expression of MjLectin in the hepatopancreas was up-regulated significantly at 48 h (P<0.01) compared with the control group. These results suggested that MjLectin might be involved in the innate immune defense against WSSV infection.

  7. C-type lectin receptors and RIG-I-like receptors: new points on the oncogenomics map

    International Nuclear Information System (INIS)

    The group of pattern recognition receptors includes families of Toll-like receptors, NOD-like receptors, C-type lectin receptors, and RIG-I-like receptors. They are key sensors for a number of infectious agents, some of which are oncogenic, and they launch an immune response against them, normally promoting their eradication. Inherited variations in genes encoding these receptors and proteins and their signaling pathways may affect their function, possibly modulating cancer risk and features of cancer progression. There are numerous studies investigating the association of single nucleotide polymorphisms within or near genes encoding Toll-like receptors and NOD-like receptors, cancer risk, and features of cancer progression. However, there is an almost total absence of articles analyzing the correlation between polymorphisms of genes encoding C-type lectin receptors and RIG-I-like receptors and cancer risk or progression. Nevertheless, there is some evidence supporting the hypothesis that inherited C-type lectin receptor and RIG-I-like receptor variants can be associated with increased cancer risk. Certain C-type lectin receptors and RIG-I-like receptors recognize pathogen-associated molecular patterns of potentially oncogenic infectious agents, and certain polymorphisms of genes encoding C-type lectin receptors and RIG-I-like receptors may have functional consequences at the molecular level that can lead to association of such single nucleotide polymorphisms with risk or progression of some diseases that may modulate cancer risk, so these gene polymorphisms may affect cancer risk indirectly. Polymorphisms of genes encoding C-type lectin receptors and RIG-I-like receptors thereby may be correlated with a risk of lung, oral, esophageal, gastric, colorectal, and liver cancer, as well as nasopharyngeal carcinoma, glioblastoma, multiple myeloma, and lymphoma. The list of the most promising polymorphisms for oncogenomic investigations may include rs1926736, rs2478577

  8. Critical roles of sea cucumber C-type lectin in non-self recognition and bacterial clearance.

    Science.gov (United States)

    Wei, Xiumei; Liu, Xiangquan; Yang, Jianmin; Wang, Sheng; Sun, Guohua; Yang, Jialong

    2015-08-01

    C-type lectin is one important pattern recognition receptor (PRR) that plays crucial roles in multiple immune responses. A C-type lectin from sea cucumber Apostichopus japonicus (AjCTL-1) was characterized in the present study. The amino acid sequence of AjCTL-1 shared high similarities with other C-type lectins from invertebrates and vertebrates. The C-type lectin domain (CTLD) of AjCTL-1 contained a Ca(2+)-binding site 2 and four conserved cysteine residues. AjCTL-1 mRNA expression patterns in tissues and after bacterial challenge were then analysed. Quantitative PCR revealed that AjCTL-1 mRNA was widely expressed in the tested tissues of healthy sea cucumber. The highest expression level occurred in gonad followed by body wall, coelomocytes, tentacle, intestinum and longitudinal muscle, and the lowest expression level was in respiratory tree. AjCTL-1 mRNA expression in coelomocytes was significantly induced by gram-negative Listonella anguillarum and gram-positive Micrococcus luteus, with different up-regulation patterns post-challenge. Recombinant AjCTL-1 exhibited the ability to bind peptidoglycan directly, agglutinate M. luteus, Staphylococcus aureus and Escherichia coli, in a Ca(2+)-dependant manner, and enhance the phagocytosis of coelomocytes against E. coli in vitro. The results indicated that AjCTL-1 could act as a PRR in Apostichopus japonicus and had critical roles in non-self recognition and bacterial clearance against invading microbes. PMID:26052017

  9. The Macrophage Galactose-Type C-Type Lectin (MGL Modulates Regulatory T Cell Functions.

    Directory of Open Access Journals (Sweden)

    Ilaria Grazia Zizzari

    Full Text Available Regulatory T cells (Tregs are physiologically designed to prevent autoimmune disease and maintain self-tolerance. In tumour microenvironments, their presence is related to a poor prognosis, and they influence the therapeutic outcome due to their capacity to suppress the immune response by cell-cell contact and to release immunosuppressive cytokines. In this study, we demonstrate that Treg immunosuppressive activity can be modulated by the cross-linking between the CD45RA expressed by Tregs and the C-type lectin MGL. This specific interaction strongly decreases the immunosuppressive activity of Tregs, restoring the proliferative capacity of co-cultured T lymphocytes. This effect can be attributed to changes in CD45RA and TCR signalling through the inhibition of Lck and inactivation of Zap-70, an increase in the Foxp3 methylation status and, ultimately, the reduced production of suppressive cytokines. These results indicate a role of MGL as an immunomodulator within the tumour microenvironment interfering with Treg functions, suggesting its possible use in the design of anticancer vaccines.

  10. Signalling versatility following self and non-self sensing by myeloid C-type lectin receptors

    Science.gov (United States)

    Iborra, Salvador; Sancho, David

    2015-01-01

    Among myeloid immune receptors, C-type lectin receptors (CLRs) have a remarkable capacity to sense a variety of self and non-self ligands. The coupling of CLRs to different signal transduction modules is influenced not only by the receptor, but also by the nature, density and architecture of the ligand, which can affect the rate of receptor internalization and trafficking to diverse intracellular compartments. Understanding how the variety of self and non-self ligands triggers differential CLR signalling and function presents a fascinating biological challenge. Non-self ligands usually promote inflammation and immunity, whereas self ligands are frequently involved in communication and tolerance. But pathogens can mimic self-inhibitory signals to escape immune surveillance, and endogenous ligands can contribute to the sensing of pathogens through CLRs. In this review, we survey the complexity and flexibility in functional outcome found in the myeloid CLRs, which is not only based on their differing intracellular motifs, but is also conditioned by the physical nature, affinity and avidity of the ligand. PMID:25269828

  11. Fungal engagement of the C-type lectin mincle suppresses dectin-1-induced antifungal immunity.

    Science.gov (United States)

    Wevers, Brigitte A; Kaptein, Tanja M; Zijlstra-Willems, Esther M; Theelen, Bart; Boekhout, Teun; Geijtenbeek, Teunis B H; Gringhuis, Sonja I

    2014-04-01

    Recognition of fungal pathogens by C-type lectin receptor (CLR) dectin-1 on human dendritic cells is essential for triggering protective antifungal TH1 and TH17 immune responses. We show that Fonsecaea monophora, a causative agent of chromoblastomycosis, a chronic fungal skin infection, evades these antifungal responses by engaging CLR mincle and suppressing IL-12, which drives TH1 differentiation. Dectin-1 triggering by F. monophora activates transcription factor IRF1, which is crucial for IL12A transcription via nucleosome remodeling. However, simultaneous F. monophora binding to mincle induces an E3 ubiquitin ligase Mdm2-dependent degradation pathway, via Syk-CARD9-mediated PKB signaling, that leads to loss of nuclear IRF1 activity, hence blocking IL12A transcription. The absence of IL-12 leads to impaired TH1 responses and promotes TH2 polarization. Notably, mincle is similarly exploited by other chromoblastomycosis-associated fungi to redirect TH responses. Thus, mincle is a fungal receptor that can suppress antifungal immunity and, as such, is a potential therapeutic target. PMID:24721577

  12. Differential expression of skin mucus C-type lectin in two freshwater eel species, Anguilla marmorata and Anguilla japonica.

    Science.gov (United States)

    Tsutsui, Shigeyuki; Yoshinaga, Tatsuki; Komiya, Kaoru; Yamashita, Hiroka; Nakamura, Osamu

    2016-08-01

    Two types of lactose-specific lectins, galectin (AJL-1) and C-type lectin (AJL-2), were previously identified in the mucus of adult Anguilla japonica. Here, we compared the expression profiles of these two homologous lectins at the adult and juvenile stages between the tropical eel Anguilla marmorata and the temperate eel A. japonica. Only one lectin, predicted to be an orthologue of AJL-1 by LC-MS/MS, was detected in the mucus of adult A. marmorata. We also found that an orthologous gene to AJL-2 was expressed at very low levels, or not at all, in the skin of adult A. marmorata. However, we detected the gene expression of an AJL-2-orthologue in the skin of juvenile A. marmorata, and a specific antibody also detected the lectin in the juvenile fish epidermis. These findings suggest that expression profiles of mucosal lectins vary during development as well as between species in the Anguilla genus. PMID:27026508

  13. C-type Lectin Receptor Expression on Human Basophils and Effects of Allergen-Specific Immunotherapy.

    Science.gov (United States)

    Lundberg, K; Rydnert, F; Broos, S; Andersson, M; Greiff, L; Lindstedt, M

    2016-09-01

    Basophils are emerging as immunoregulatory cells capable of interacting with their environment not only via their characteristic IgE-mediated activation, but also in an IgE-independent manner. Basophils are known to express and respond to stimulation via TLR2, TLR4, DC-SIGN and DCIR, but whether basophils also express other C-type lectin receptors (CLRs) is largely unknown. In this study, we investigate the CLR expression profile of human basophils using multicolour flow cytometry. As FcRs as well as some CLRs are associated with allergen recognition and shown to be involved in subsequent immune responses, the expression of CLRs and FcRs on peripheral blood basophils, as well as their frequency, was monitored for 1 year in subjects undergoing subcutaneous allergen-specific immunotherapy (AIT). Here, we show that human basophils express CLECSF14, DEC205, Dectin-1, Dectin-2 and MRC2. Furthermore, we demonstrate that the frequencies of basophils expressing the allergy-associated CLRs Dectin-1 and Dectin-2 were significantly reduced after 1 year and 8 weeks of AIT, respectively. In contrast, the frequency of basophils positive for FcγRII, as well as the fraction of total basophils, significantly increased after 1 year of AIT. The herein demonstrated expression of various CLRs on basophils, and their altered CLR and FcR expression profile upon AIT, suggest yet unexplored ways by which basophils can interact with antigens and may point to novel immunoregulatory functions targeted through AIT. PMID:27354239

  14. Hemolytic C-Type Lectin CEL-III from Sea Cucumber Expressed in Transgenic Mosquitoes Impairs Malaria Parasite Development

    OpenAIRE

    Shigeto Yoshida; Yohei Shimada; Daisuke Kondoh; Yoshiaki Kouzuma; Ghosh, Anil K.; Marcelo Jacobs-Lorena; Sinden, Robert E.

    2007-01-01

    The midgut environment of anopheline mosquitoes plays an important role in the development of the malaria parasite. Using genetic manipulation of anopheline mosquitoes to change the environment in the mosquito midgut may inhibit development of the malaria parasite, thus blocking malaria transmission. Here we generate transgenic Anopheles stephensi mosquitoes that express the C-type lectin CEL-III from the sea cucumber, Cucumaria echinata, in a midgut-specific manner. CEL-III has strong and ra...

  15. Mechanistic insights into the role of C-type lectin receptor/CARD9 signaling in human antifungal immunity

    OpenAIRE

    Drummond, Rebecca A.; Lionakis, Michail S.

    2016-01-01

    Human CARD9 deficiency is an autosomal recessive primary immunodeficiency disorder caused by biallelic mutations in the gene CARD9, which encodes a signaling protein that is found downstream of many C-type lectin receptors (CLRs). CLRs encompass a large family of innate recognition receptors, expressed predominantly by myeloid and epithelial cells, which bind fungal carbohydrates and initiate antifungal immune responses. Accordingly, human CARD9 deficiency is associated with the spontaneous d...

  16. C-type lectin receptor-induced NF-κB activation in innate immune and inflammatory responses

    OpenAIRE

    Kingeter, Lara M.; Lin, Xin

    2012-01-01

    The C-type lectin receptors (CLRs) belong to a large family of proteins that contain a carbohydrate recognition domain (CRD) and calcium binding sites on their extracellular domains. Recent studies indicate that many CLRs, such as Dectin-1, Dectin-2 and Mincle, function as pattern recognition receptors (PRRs) recognizing carbohydrate ligands from infected microorganisms. Upon ligand binding, these CLRs induce multiple signal transduction cascades through their own immunoreceptor tyrosine-base...

  17. Mechanistic Insights into the Role of C-Type Lectin Receptor/CARD9 Signaling in Human Antifungal Immunity

    OpenAIRE

    Drummond, Rebecca A.; Lionakis, Michail S.

    2016-01-01

    Human CARD9 deficiency is an autosomal recessive primary immunodeficiency disorder caused by biallelic mutations in the gene CARD9, which encodes a signaling protein that is found downstream of many C-type lectin receptors (CLRs). CLRs encompass a large family of innate recognition receptors, expressed predominantly by myeloid and epithelial cells, which bind fungal carbohydrates and initiate antifungal immune responses. Accordingly, human CARD9 deficiency is associated with the spontaneous d...

  18. C-type lectin receptors differentially induce Th17 cells and vaccine immunity to the endemic mycosis of North America

    OpenAIRE

    Wang, Huafeng; LeBert, Vanessa; Hung, Chiung Yu; Galles, Kevin; Saijo, Shinobu; Lin, Xin; Cole, Garry T.; Bruce S Klein; Wüthrich, Marcel

    2014-01-01

    Vaccine immunity to the endemic mycoses of North America requires Th17 cells, but the pattern recognition receptors and signaling pathways that drive these protective responses have not been defined. We show that C-type lectin receptors (CLRs) exert divergent contributions to the development of anti-fungal Th17 cells and vaccine resistance against Blastomyces dermatitidis, Histoplasma capsulatum and Coccidioides posadasii. Acquired immunity to B. dermatitidis requires Dectin-2, whereas vaccin...

  19. Differential Use of the C-Type Lectins L-SIGN and DC-SIGN for Phlebovirus Endocytosis.

    Science.gov (United States)

    Léger, Psylvia; Tetard, Marilou; Youness, Berthe; Cordes, Nicole; Rouxel, Ronan N; Flamand, Marie; Lozach, Pierre-Yves

    2016-06-01

    Bunyaviruses represent a growing threat to humans and livestock globally. The receptors, cellular factors and endocytic pathways used by these emerging pathogens to infect cells remain largely unidentified and poorly characterized. DC-SIGN is a C-type lectin highly expressed on dermal dendritic cells that has been found to act as an authentic entry receptor for many phleboviruses (Bunyaviridae), including Rift Valley fever virus (RVFV), Toscana virus (TOSV) and Uukuniemi virus (UUKV). We found that these phleboviruses can exploit another C-type lectin, L-SIGN, for infection. L-SIGN shares 77% sequence homology with DC-SIGN and is expressed on liver sinusoidal endothelial cells. L-SIGN is required for UUKV binding but not for virus internalization. An endocytosis-defective mutant of L-SIGN was still able to mediate virus uptake and infection, indicating that L-SIGN acts as an attachment receptor for phleboviruses rather than an endocytic receptor. Our results point out a fundamental difference in the use of the C-type lectins L-SIGN and DC-SIGN by UUKV to enter cells, although both proteins are closely related in terms of molecular structure and biological function. This study sheds new light on the molecular mechanisms by which phleboviruses target the liver and also highlights the added complexity in virus-receptor interactions beyond attachment. PMID:26990254

  20. Galactose recognition by a tetrameric C-type lectin, CEL-IV, containing the EPN carbohydrate recognition motif.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Kamiya, Takuro; Kusunoki, Masami; Nakamura-Tsuruta, Sachiko; Hirabayashi, Jun; Goda, Shuichiro; Unno, Hideaki

    2011-03-25

    CEL-IV is a C-type lectin isolated from a sea cucumber, Cucumaria echinata. This lectin is composed of four identical C-type carbohydrate-recognition domains (CRDs). X-ray crystallographic analysis of CEL-IV revealed that its tetrameric structure was stabilized by multiple interchain disulfide bonds among the subunits. Although CEL-IV has the EPN motif in its carbohydrate-binding sites, which is known to be characteristic of mannose binding C-type CRDs, it showed preferential binding of galactose and N-acetylgalactosamine. Structural analyses of CEL-IV-melibiose and CEL-IV-raffinose complexes revealed that their galactose residues were recognized in an inverted orientation compared with mannose binding C-type CRDs containing the EPN motif, by the aid of a stacking interaction with the side chain of Trp-79. Changes in the environment of Trp-79 induced by binding to galactose were detected by changes in the intrinsic fluorescence and UV absorption spectra of WT CEL-IV and its site-directed mutants. The binding specificity of CEL-IV toward complex oligosaccharides was analyzed by frontal affinity chromatography using various pyridylamino sugars, and the results indicate preferential binding to oligosaccharides containing Galβ1-3/4(Fucα1-3/4)GlcNAc structures. These findings suggest that the specificity for oligosaccharides may be largely affected by interactions with amino acid residues in the binding site other than those determining the monosaccharide specificity. PMID:21247895

  1. Glycodendrimeric ligands of c-type lectin receptors as ther agents in experimental cancer

    Czech Academy of Sciences Publication Activity Database

    Pospíšil, Miloslav; Vannucci, Luca; Fišerová, Anna; Krausová, Kateřina; Horváth, Ondřej; Křen, Vladimír; Lindhorst, T.; Sadalapure, K.; Bezouška, Karel

    2001-01-01

    Roč. 495, - (2001), s. 343-348. ISSN 0065-2598 R&D Projects: GA ČR GV312/98/K034; GA AV ČR IAA7020006 Keywords : introduction * lectin * receptors Subject RIV: EE - Microbiology, Virology Impact factor: 0.513, year: 2000

  2. Bilinexin, a snake C-type lectin from Agkistrodon bilineatus venom agglutinates platelets via GPIb and alpha2beta1.

    Science.gov (United States)

    Du, X Y; Navdaev, A; Clemetson, J M; Magnenat, E; Wells, T N; Clemetson, K J

    2001-11-01

    A new snake protein, named bilinexin, has been purified from Agkistrodon bilineatus venom by ion-exchange chromatography and gel filtration chromatography. Under non-reducing conditions it has a mass of 110 kDa protein on SDS-PAGE. On reduction, it can be separated into five subunits with masses in the range 13-25 kDa. The N-terminal sequences of these subunits are very similar to those of convulxin or the alboaggregins, identifying bilinexin as a new member of the snake C-type lectin family, unusual in having multiple subunits. Bilinexin agglutinates fixed platelets. washed platelets and platelet rich plasma (PRP) without obvious activation (shape change) as confirmed by light microscope examination. Both inhibitory and binding studies indicate that antibodies against alpha2beta1 inhibit not only platelet agglutination induced by bilinexin, but also bilinexin binding to platelets. VM16d, a monoclonal anti-GPIbalpha antibody, completely inhibits platelet agglutination induced by bilinexin, and polyclonal antibodies against GPIbalpha prevent its binding to platelets. However, neither convulxin, polyclonal anti-GPVI antibodies, nor GPIIb/IIIa inhibitors affect its binding to and agglutination of platelets. Bilinexin neither activates GPIIb/IIIa integrin on platelets nor induces tyrosine phosphorylation of platelet proteins, nor increases intracellular Ca2+ in platelets. Like alboaggregin B, bilinexin agglutinates platelets, which makes it a good tool to investigate the differences in mechanism between snake C-type lectins causing platelet agglutination and those that induce full activation. PMID:11816718

  3. C-type lectin receptors differentially induce th17 cells and vaccine immunity to the endemic mycosis of North America.

    Science.gov (United States)

    Wang, Huafeng; LeBert, Vanessa; Hung, Chiung Yu; Galles, Kevin; Saijo, Shinobu; Lin, Xin; Cole, Garry T; Klein, Bruce S; Wüthrich, Marcel

    2014-02-01

    Vaccine immunity to the endemic mycoses of North America requires Th17 cells, but the pattern recognition receptors and signaling pathways that drive these protective responses have not been defined. We show that C-type lectin receptors exert divergent contributions to the development of antifungal Th17 cells and vaccine resistance against Blastomyces dermatitidis, Histoplasma capsulatum, and Coccidioides posadasii. Acquired immunity to B. dermatitidis requires Dectin-2, whereas vaccination against H. capsulatum and C. posadasii infection depends on innate sensing by Dectin-1 and Dectin-2, but not Mincle. Tracking Ag-specific T cells in vivo established that the Card9 signaling pathway acts indispensably and exclusively on differentiation of Th17 cells, while leaving intact their activation, proliferation, survival, and migration. Whereas Card9 signaling is essential, C-type lectin receptors offer distinct and divergent contributions to vaccine immunity against these endemic fungal pathogens. Our work provides new insight into innate immune mechanisms that drive vaccine immunity and Th17 cells. PMID:24391211

  4. Mannose-recognition mutant of the galactose/N-acetylgalactosamine-specific C-type lectin CEL-I engineered by site-directed mutagenesis

    OpenAIRE

    Moriuchi, Hiromi; Unno, Hideaki; Goda, Shuichiro; Tateno, Hiroaki; Hirabayashi, Jun; Hatakeyama, Tomomitsu

    2015-01-01

    Background CEL-I is a galactose/N-acetylgalactosamine-specific C-type lectin isolated from the sea cucumber Cucumaria echinata. Its carbohydrate-binding site contains a QPD (Gln-Pro-Asp) motif, which is generally recognized as the galactose specificity-determining motif in the C-type lectins. In our previous study, replacement of the QPD motif by an EPN (Glu-Pro-Asn) motif led to a weak binding affinity for mannose. Therefore, we examined the effects of an additional mutation in the carbohydr...

  5. C-type lectin-like carbohydrate recognition of the hemolytic lectin CEL-III containing ricin-type -trefoil folds.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Unno, Hideaki; Kouzuma, Yoshiaki; Uchida, Tatsuya; Eto, Seiichiro; Hidemura, Haruki; Kato, Norihisa; Yonekura, Masami; Kusunoki, Masami

    2007-12-28

    CEL-III is a Ca(2+)-dependent hemolytic lectin, isolated from the marine invertebrate Cucumaria echinata. The three-dimensional structure of CEL-III/GalNAc and CEL-III/methyl alpha-galactoside complexes was solved by x-ray crystallographic analysis. In these complexes, five carbohydrate molecules were found to be bound to two carbohydrate-binding domains (domains 1 and 2) located in the N-terminal 2/3 portion of the polypeptide and that contained beta-trefoil folds similar to ricin B-chain. The 3-OH and 4-OH of bound carbohydrate molecules were coordinated with Ca(2+) located at the subdomains 1alpha, 1gamma, 2alpha, 2beta, and 2gamma, simultaneously forming hydrogen bond networks with nearby amino acid side chains, which is similar to carbohydrate binding in C-type lectins. The binding of carbohydrates was further stabilized by aromatic amino acid residues, such as tyrosine and tryptophan, through a stacking interaction with the hydrophobic face of carbohydrates. The importance of amino acid residues in the carbohydrate-binding sites was confirmed by the mutational analyses. The orientation of bound GalNAc and methyl alpha-galactoside was similar to the galactose moiety of lactose bound to the carbohydrate-binding site of the ricin B-chain, although the ricin B-chain does not require Ca(2+) ions for carbohydrate binding. The binding of the carbohydrates induced local structural changes in carbohydrate-binding sites in subdomains 2alpha and 2beta. Binding of GalNAc also induced a slight change in the main chain structure of domain 3, which could be related to the conformational change upon binding of specific carbohydrates to induce oligomerization of the protein. PMID:17977832

  6. cDNA cloning,sequence analysis,and recombinant expression of akitonin beta,a C-type lectin-like protein from Agkistrodon acutus

    Institute of Scientific and Technical Information of China (English)

    Xiang-dong ZHA; Jing LIU; Kang-sen XU

    2004-01-01

    AIM: To clone the cDNA of a new member of snake venom C-type lectin-like proteins, to study its structurefunction relationships and to achieve its recombinant production. METHODS: PCR primers were designed based on the homology and cDNA was amplified by RT-PCR using total RNA from snake venom gland as the template.The PCR products were cloned into the plasmid pGEM-T and sequenced. The deduced protein sequence was analyzed with some bioinformatic programs. A recombinant expression plasmid was constructed using pBADTOPO as vector and transformed into E. coli TOP10 competent cells. RESULTS: A novel cDNA sequence encoding akitonin β was found and accepted by GenBank (accession number AF387100). Akitonin β consists of a typical carbohydrate recognition domain (CRD) of C-type lectins, and it is homologous with other snake venom C-type lectin-like proteins. It was predicted to be a platelet antagonist. Upon induction with arabinose rAkitonin β expressing in E coli was achieved at a high level (superior to 150 mg/L). The recombinant fusion protein exhibited inhibitory activities on rat platelet aggregation in vitro. CONCLUSION: A new member of snake venom C-type lectin-like proteins was discovered and characterized, and an efficient recombinant expression system was established for its production.

  7. The dendritic cell-specific C-type lectin DC-SIGN is a receptor for Schistosoma mansoni egg antigens and recognizes the glycan antigen Lewis x.

    NARCIS (Netherlands)

    Die, van I.M.; Vliet, van SJ; Nyame, AK; Cummings, RD; Bank, CM; Appelmelk, B.J.; Geijtenbeek, T.B.H.; Kooijk, van Y.

    2003-01-01

    Schistosoma mansoni soluble egg antigens (SEAs) are crucially involved in modulating the host immune response to infection by S. mansoni. We report that human dendritic cells bind SEAs through the C-type lectin dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN). Monoclonal antibodies agai

  8. Hemolytic C-type lectin CEL-III from sea cucumber expressed in transgenic mosquitoes impairs malaria parasite development.

    Directory of Open Access Journals (Sweden)

    Shigeto Yoshida

    2007-12-01

    Full Text Available The midgut environment of anopheline mosquitoes plays an important role in the development of the malaria parasite. Using genetic manipulation of anopheline mosquitoes to change the environment in the mosquito midgut may inhibit development of the malaria parasite, thus blocking malaria transmission. Here we generate transgenic Anopheles stephensi mosquitoes that express the C-type lectin CEL-III from the sea cucumber, Cucumaria echinata, in a midgut-specific manner. CEL-III has strong and rapid hemolytic activity toward human and rat erythrocytes in the presence of serum. Importantly, CEL-III binds to ookinetes, leading to strong inhibition of ookinete formation in vitro with an IC(50 of 15 nM. Thus, CEL-III exhibits not only hemolytic activity but also cytotoxicity toward ookinetes. In these transgenic mosquitoes, sporogonic development of Plasmodium berghei is severely impaired. Moderate, but significant inhibition was found against Plasmodium falciparum. To our knowledge, this is the first demonstration of stably engineered anophelines that affect the Plasmodium transmission dynamics of human malaria. Although our laboratory-based research does not have immediate applications to block natural malaria transmission, these findings have significant implications for the generation of refractory mosquitoes to all species of human Plasmodium and elucidation of mosquito-parasite interactions.

  9. Hemolytic C-type lectin CEL-III from sea cucumber expressed in transgenic mosquitoes impairs malaria parasite development.

    Science.gov (United States)

    Yoshida, Shigeto; Shimada, Yohei; Kondoh, Daisuke; Kouzuma, Yoshiaki; Ghosh, Anil K; Jacobs-Lorena, Marcelo; Sinden, Robert E

    2007-12-01

    The midgut environment of anopheline mosquitoes plays an important role in the development of the malaria parasite. Using genetic manipulation of anopheline mosquitoes to change the environment in the mosquito midgut may inhibit development of the malaria parasite, thus blocking malaria transmission. Here we generate transgenic Anopheles stephensi mosquitoes that express the C-type lectin CEL-III from the sea cucumber, Cucumaria echinata, in a midgut-specific manner. CEL-III has strong and rapid hemolytic activity toward human and rat erythrocytes in the presence of serum. Importantly, CEL-III binds to ookinetes, leading to strong inhibition of ookinete formation in vitro with an IC(50) of 15 nM. Thus, CEL-III exhibits not only hemolytic activity but also cytotoxicity toward ookinetes. In these transgenic mosquitoes, sporogonic development of Plasmodium berghei is severely impaired. Moderate, but significant inhibition was found against Plasmodium falciparum. To our knowledge, this is the first demonstration of stably engineered anophelines that affect the Plasmodium transmission dynamics of human malaria. Although our laboratory-based research does not have immediate applications to block natural malaria transmission, these findings have significant implications for the generation of refractory mosquitoes to all species of human Plasmodium and elucidation of mosquito-parasite interactions. PMID:18159942

  10. Conservation of the C-type lectin fold for massive sequence variation in a Treponema diversity-generating retroelement

    Energy Technology Data Exchange (ETDEWEB)

    Le Coq, Johanne; Ghosh, Partho (UCSD)

    2012-06-19

    Anticipatory ligand binding through massive protein sequence variation is rare in biological systems, having been observed only in the vertebrate adaptive immune response and in a phage diversity-generating retroelement (DGR). Earlier work has demonstrated that the prototypical DGR variable protein, major tropism determinant (Mtd), meets the demands of anticipatory ligand binding by novel means through the C-type lectin (CLec) fold. However, because of the low sequence identity among DGR variable proteins, it has remained unclear whether the CLec fold is a general solution for DGRs. We have addressed this problem by determining the structure of a second DGR variable protein, TvpA, from the pathogenic oral spirochete Treponema denticola. Despite its weak sequence identity to Mtd ({approx}16%), TvpA was found to also have a CLec fold, with predicted variable residues exposed in a ligand-binding site. However, this site in TvpA was markedly more variable than the one in Mtd, reflecting the unprecedented approximate 10{sup 20} potential variability of TvpA. In addition, similarity between TvpA and Mtd with formylglycine-generating enzymes was detected. These results provide strong evidence for the conservation of the formylglycine-generating enzyme-type CLec fold among DGRs as a means of accommodating massive sequence variation.

  11. The Structure of the Poxvirus A33 Protein Reveals a Dimer of Unique C-Type Lectin-Like Domains

    Energy Technology Data Exchange (ETDEWEB)

    Su, Hua-Poo; Singh, Kavita; Gittis, Apostolos G.; Garboczi, David N. (NIH)

    2010-11-03

    The current vaccine against smallpox is an infectious form of vaccinia virus that has significant side effects. Alternative vaccine approaches using recombinant viral proteins are being developed. A target of subunit vaccine strategies is the poxvirus protein A33, a conserved protein in the Chordopoxvirinae subfamily of Poxviridae that is expressed on the outer viral envelope. Here we have determined the structure of the A33 ectodomain of vaccinia virus. The structure revealed C-type lectin-like domains (CTLDs) that occur as dimers in A33 crystals with five different crystal lattices. Comparison of the A33 dimer models shows that the A33 monomers have a degree of flexibility in position within the dimer. Structural comparisons show that the A33 monomer is a close match to the Link module class of CTLDs but that the A33 dimer is most similar to the natural killer (NK)-cell receptor class of CTLDs. Structural data on Link modules and NK-cell receptor-ligand complexes suggest a surface of A33 that could interact with viral or host ligands. The dimer interface is well conserved in all known A33 sequences, indicating an important role for the A33 dimer. The structure indicates how previously described A33 mutations disrupt protein folding and locates the positions of N-linked glycosylations and the epitope of a protective antibody.

  12. Mechanistic Insights into the Role of C-Type Lectin Receptor/CARD9 Signaling in Human Antifungal Immunity.

    Science.gov (United States)

    Drummond, Rebecca A; Lionakis, Michail S

    2016-01-01

    Human CARD9 deficiency is an autosomal recessive primary immunodeficiency disorder caused by biallelic mutations in the gene CARD9, which encodes a signaling protein that is found downstream of many C-type lectin receptors (CLRs). CLRs encompass a large family of innate recognition receptors, expressed predominantly by myeloid and epithelial cells, which bind fungal carbohydrates and initiate antifungal immune responses. Accordingly, human CARD9 deficiency is associated with the spontaneous development of persistent and severe fungal infections that primarily localize to the skin and subcutaneous tissue, mucosal surfaces and/or central nervous system (CNS). In the last 3 years, more than 15 missense and nonsense CARD9 mutations have been reported which associate with the development of a wide spectrum of fungal infections caused by a variety of fungal organisms. The mechanisms by which CARD9 provides organ-specific protection against these fungal infections are now emerging. In this review, we summarize recent immunological and clinical advances that have provided significant mechanistic insights into the pathogenesis of human CARD9 deficiency. We also discuss how genetic mutations in CARD9-coupled receptors (Dectin-1, Dectin-2) and CARD9-binding partners (MALT1, BCL10) affect human antifungal immunity relative to CARD9 deficiency, and we highlight major understudied research questions which merit future investigation. PMID:27092298

  13. Mechanistic insights into the role of C-type lectin receptor/CARD9 signaling in human antifungal immunity

    Directory of Open Access Journals (Sweden)

    Rebecca A. Drummond

    2016-04-01

    Full Text Available Human CARD9 deficiency is an autosomal recessive primary immunodeficiency disorder caused by biallelic mutations in the gene CARD9, which encodes a signaling protein that is found downstream of many C-type lectin receptors (CLRs. CLRs encompass a large family of innate recognition receptors, expressed predominantly by myeloid and epithelial cells, which bind fungal carbohydrates and initiate antifungal immune responses. Accordingly, human CARD9 deficiency is associated with the spontaneous development of persistent and severe fungal infections that primarily localize to the skin and subcutaneous tissue, mucosal surfaces and/or central nervous system (CNS. In the last few years, more than 15 missense and nonsense CARD9 mutations have been reported which associate with the development of a wide spectrum of fungal infections caused by a variety of fungal organisms. The mechanisms by which CARD9 provides organ-specific protection against these fungal infections are now emerging. In this review, we summarize recent immunological and clinical advances that have provided significant mechanistic insights into the pathogenesis of human CARD9 deficiency. We also discuss how genetic mutations in CARD9-coupled receptors (Dectin-1, Dectin-2 and CARD9-binding partners (MALT1, BCL10 affect human antifungal immunity relative to CARD9 deficiency, and we highlight major understudied research questions which merit future investigation.

  14. Crystallization and preliminary crystallographic study of an invertebrate C-type lectin, CEL-I, from the marine invertebrate Cucumaria echinata.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Matsuo, Noriaki; Aoyagi, Haruhiko; Sugawara, Hajime; Uchida, Tatsuya; Kurisu, Genji; Kusunoki, Masami

    2002-01-01

    CEL-I is a GalNAc-specific carbohydrate-binding protein (lectin) isolated from the sea cucumber Cucumaria echinata. This protein belongs to the widely distributed C-type lectin family of animal lectins, which require Ca(2+) for their carbohydrate-binding ability and play important roles in various molecular-recognition processes in organisms. CEL-I was crystallized with 2-methyl-2,4-pentanediol using the hanging-drop vapour-diffusion technique. The CEL-I crystals belong to the monoclinic space group C2, with unit-cell parameters a = 92.38 (3), b = 69.94 (3), c = 76.69 (3) A, beta = 136.46 (2) degrees. Diffraction data were collected to 2.0 A resolution using synchrotron radiation. The asymmetric unit contains one CEL-I molecule. PMID:11752793

  15. Mitogenic activity of CEL-I, an N-acetylgalactosamine (GalNAc)-specific C-type lectin, isolated from the marine invertebrate Cucumaria echinata (Holothuroidea).

    Science.gov (United States)

    Jiang, Zedong; Kim, Daekyung; Yamasaki, Yasuhiro; Yamanishi, Tomohiro; Hatakeyama, Tomomitsu; Yamaguchi, Kenichi; Oda, Tatsuya

    2010-01-01

    An N-acetylgalactosamine (GalNAc)-specific Ca(2+)-dependent lectin (C-type lectin), isolated from the marine invertebrate Holothuroidea (Cucumaria echinata), CEL-I, showed potent mitogenic activity toward normal mouse spleen cells. The mitogenic activity of CEL-I, which reached a maximum at 100 microg/ml, was inhibited by GalNAc in a concentration-dependent manner. The mitogenic effect of CEL-I at 10 microg/ml on T cell- enriched splenocytes was at a similar level due to a well-known T cell mitogen, concanavalin A (Con A), at 10 microg/ml. Furthermore, CEL-I evoked a mitogenic response from nude mouse spleen cells, while no significant effects of Con A on this cell population were observed over a wide range of concentrations. These results suggest that CEL-I is a potent mitogenic lectin with the ability to stimulate both T and B cells. PMID:20699569

  16. Syk Kinase-Coupled C-type Lectin Receptors Engage Protein Kinase C-δ to Elicit Card9 Adaptor-Mediated Innate Immunity

    OpenAIRE

    Strasser, Dominikus; Neumann, Konstantin; Bergmann, Hanna; Marakalala, Mohlopheni J.; Guler, Reto; Rojowska, Anna; Hopfner, Karl-Peter; Brombacher, Frank; Urlaub, Henning; Baier, Gottfried; Brown, Gordon D.; Leitges, Michael; Ruland, Jürgen

    2012-01-01

    Summary C-type lectin receptors (CLRs) that couple with the kinase Syk are major pattern recognition receptors for the activation of innate immunity and host defense. CLRs recognize fungi and other forms of microbial or sterile danger, and they induce inflammatory responses through the adaptor protein Card9. The mechanisms relaying CLR proximal signals to the core Card9 module are unknown. Here we demonstrated that protein kinase C-δ (PKCδ) was activated upon Dectin-1-Syk signaling, mediated ...

  17. CEL-I, an N-acetylgalactosamine (GalNAc)-specific C-type lectin, induces nitric oxide production in RAW264.7 mouse macrophage cell line.

    OpenAIRE

    Yamanishi, Tomohiro; Hatakeyama, Tomomitsu; YAMAGUCHI, Kenichi; Oda, Tatsuya

    2009-01-01

    We found that CEL-I, a GalNAc-specific C-type lectin isolated from the marine invertebrate Holothuroidea (Cucumaria echinata), induces inducible nitric oxide synthase (iNOS) expression and NO production in RAW264.7 cells. The NO production was inhibited by an iNOS inhibitor, L-NAME, but was not by a lipopolysaccharide (LPS) inhibitor, polymyxin B. In the presence of 0.1-M GalNAc, increased NO production by CEL-I-treated RAW264.7 cells was observed rather than the inhibition. Bovine serum albu...

  18. Recombinant Expression, In Vitro Refolding and Characterizing Disulfide Bonds of a Mouse Inhibitory C-Type Lectin-Like Receptor Nkrp1b

    Czech Academy of Sciences Publication Activity Database

    Hernychová, Lucie; Mrázek, Hynek; Ivanova, Ljubina; Kukačka, Zdeněk; Chmelík, Josef; Novák, Petr

    2015-01-01

    Roč. 64, č. 2015 (2015), s. 85-93. ISSN 0862-8408 R&D Projects: GA MŠk(CZ) EE2.3.30.0003; GA MŠk(CZ) ED1.1.00/02.0109; GA MŠk LO1509 Grant ostatní: OPPC(XE) CZ.2.16/3.1.00/24023 Institutional support: RVO:61388971 Keywords : NK cell * C-type lectin-like receptor (CTLR) * Nkrp1 Subject RIV: CE - Biochemistry Impact factor: 1.293, year: 2014

  19. Alteration of the carbohydrate-binding specificity of a C-type lectin CEL-I mutant with an EPN carbohydrate-binding motif.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Ishimine, Tomohiro; Baba, Tomohiro; Kimura, Masanari; Unno, Hideaki; Goda, Shuichiro

    2013-07-01

    CEL-I is a Gal/GalNAc-specific C-type lectin isolated from the sea cucumber Cucumaria echinata. This lectin is composed of two carbohydrate-recognition domains (CRDs) with the carbohydrate-recognition motif QPD (Gln-Pro- Asp), which is generally known to exist in galactose-specific C-type CRDs. In the present study, a mutant CEL-I with EPN (Glu-Pro-Asn) motif, which is thought to be responsible for the carbohydrate-recognition of mannose-specific Ctype CRDs, was produced in Escherichia coli, and its effects on the carbohydrate-binding specificity were examined using polyamidoamine dendrimer (PD) conjugated with carbohydrates. Although wild-type CEL-I effectively formed complexes with N-acetylgalactosamine (GalNAc)-PD but not with mannose-PD, the mutant CEL-I showed relatively weak but definite affinity for mannose-PD. These results indicated that the QPD and EPN motifs play a significant role in the carbohydrate-recognition mechanism of CEL-I, especially in the discrimination of galactose and mannose. Additional mutations in the recombinant CEL-I binding site may further increase its specificity for mannose, and should provide insights into designing novel carbohydrate-recognition proteins. PMID:23157284

  20. Evolution of the C-Type Lectin-Like Receptor Genes of the DECTIN-1 Cluster in the NK Gene Complex

    Directory of Open Access Journals (Sweden)

    Susanne Sattler

    2012-01-01

    Full Text Available Pattern recognition receptors are crucial in initiating and shaping innate and adaptive immune responses and often belong to families of structurally and evolutionarily related proteins. The human C-type lectin-like receptors encoded in the DECTIN-1 cluster within the NK gene complex contain prominent receptors with pattern recognition function, such as DECTIN-1 and LOX-1. All members of this cluster share significant homology and are considered to have arisen from subsequent gene duplications. Recent developments in sequencing and the availability of comprehensive sequence data comprising many species showed that the receptors of the DECTIN-1 cluster are not only homologous to each other but also highly conserved between species. Even in Caenorhabditis elegans, genes displaying homology to the mammalian C-type lectin-like receptors have been detected. In this paper, we conduct a comprehensive phylogenetic survey and give an up-to-date overview of the currently available data on the evolutionary emergence of the DECTIN-1 cluster genes.

  1. E3 ubiquitin ligase CHIP interacts with C-type lectin-like receptor CLEC-2 and promotes its ubiquitin-proteasome degradation.

    Science.gov (United States)

    Shao, Miaomiao; Li, Lili; Song, Shushu; Wu, Weicheng; Peng, Peike; Yang, Caiting; Zhang, Mingming; Duan, Fangfang; Jia, Dongwei; Zhang, Jie; Wu, Hao; Zhao, Ran; Wang, Lan; Ruan, Yuanyuan; Gu, Jianxin

    2016-10-01

    C-type lectin-like receptor 2 (CLEC-2) was originally identified as a member of non-classical C-type lectin-like receptors in platelets and immune cells. Activation of CLEC-2 is involved in thrombus formation, lymphatic/blood vessel separation, platelet-mediated tumor metastasis and immune response. Nevertheless, the regulation of CLEC-2 expression is little understood. In this study, we identified that the C terminus of Hsc70-interacting protein (CHIP) interacted with CLEC-2 by mass spectrometry analysis, and CHIP decreased the protein expression of CLEC-2 through lysine-48-linked ubiquitination and proteasomal degradation. Deleted and point mutation also revealed that CHIP controlled CLEC-2 protein expression via both tetratricopeptide repeats (TPR) domain and Ubox domain in a HSP70/90-independent manner. Moreover, reduced CHIP expression was associated with decreased CLEC-2 polyubiquitination and increased CLEC-2 protein levels in PMA-induced differentiation of THP-1 monocytes into macrophages. These results indicate that CLEC-2 is the target substrate of E3 ubiquitin ligase CHIP, and suggest that the CHIP/CLEC-2 axis may play an important role in the modulation of immune response. PMID:27443248

  2. The Cryptosporidium parvum C-Type Lectin CpClec Mediates Infection of Intestinal Epithelial Cells via Interactions with Sulfated Proteoglycans.

    Science.gov (United States)

    Ludington, Jacob G; Ward, Honorine D

    2016-05-01

    The apicomplexan parasite Cryptosporidium causes significant diarrheal disease worldwide. Effective anticryptosporidial agents are lacking, in part because the molecular mechanisms underlying Cryptosporidium-host cell interactions are poorly understood. Previously, we identified and characterized a novel Cryptosporidium parvum C-type lectin domain-containing mucin-like glycoprotein, CpClec. In this study, we evaluated the mechanisms underlying interactions of CpClec with intestinal epithelial cells by using an Fc-tagged recombinant protein. CpClec-Fc displayed Ca(2+)-dependent, saturable binding to HCT-8 and Caco-2 cells and competitively inhibited C. parvum attachment to and infection of HCT-8 cells. Binding of CpClec-Fc was specifically inhibited by sulfated glycosaminoglycans, particularly heparin and heparan sulfate. Binding was reduced after the removal of heparan sulfate and following the inhibition of glycosaminoglycan synthesis or sulfation in HCT-8 cells. Like CpClec-Fc binding, C. parvum attachment to and infection of HCT-8 cells were inhibited by glycosaminoglycans and were reduced after heparan sulfate removal or inhibition of glycosaminoglycan synthesis or sulfation. Lastly, CpClec-Fc binding and C. parvum sporozoite attachment were significantly decreased in CHO cell mutants defective in glycosaminoglycan synthesis. Together, these results indicate that CpClec is a novel C-type lectin that mediates C. parvum attachment and infection via Ca(2+)-dependent binding to sulfated proteoglycans on intestinal epithelial cells. PMID:26975991

  3. Characteristic recognition of N-acetylgalactosamine by an invertebrate C-type Lectin, CEL-I, revealed by X-ray crystallographic analysis.

    Science.gov (United States)

    Sugawara, Hajime; Kusunoki, Masami; Kurisu, Genji; Fujimoto, Tokiko; Aoyagi, Haruhiko; Hatakeyama, Tomomitsu

    2004-10-22

    CEL-I is a C-type lectin, purified from the sea cucumber Cucumaria echinata, that shows a high specificity for N-acetylgalactosamine (GalNAc). We determined the crystal structures of CEL-I and its complex with GalNAc at 2.0 and 1.7 A resolution, respectively. CEL-I forms a disulfide-linked homodimer and contains two intramolecular disulfide bonds, although it lacks one intramolecular disulfide bond that is widely conserved among various C-type carbohydrate recognition domains (CRDs). Although the sequence similarity of CEL-I with other C-type CRDs is low, the overall folding of CEL-I was quite similar to those of other C-type CRDs. The structure of the complex with GalNAc revealed that the basic recognition mode of GalNAc was very similar to that for the GalNAc-binding mutant of the mannose-binding protein. However, the acetamido group of GalNAc appeared to be recognized more strongly by the combination of hydrogen bonds to Arg115 and van der Waals interaction with Gln70. Mutational analyses, in which Gln70 and/or Arg115 were replaced by alanine, confirmed that these residues contributed to GalNAc recognition in a cooperative manner. PMID:15319425

  4. Characterization of a recombinant C-type lectin, rCEL-IV, expressed in Escherichia coli cells using a synthetic gene.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Hozawa, Takao; Hirotani, Iyo; Tsuda, Nobuaki; Kusunoki, Masami; Shiba, Kohei

    2006-03-01

    The body fluid of marine invertebrate Cucumaria echinata (Holothuroidea) contains four Ca2+-dependent galactose-specific lectins. One of these lectins, CEL-IV, is composed of a C-type carbohydrate-recognition domain homotetramer. CEL-IV exhibits higher specificity for alpha-galactosides than for beta-galactosides, while other C. echinata lectins show preferential binding of beta-galactosides. We constructed an artificial synthetic gene for recombinant CEL-IV (rCEL-IV) based on the amino acid sequence previously determined from the purified protein. rCEL-IV was expressed in Escherichia coli cells as inclusion bodies. After the refolding process, most of rCEL-IV spontaneously formed a homotetramer structure having interchain disulfide bonds. The secondary structure of rCEL-IV was similar to that of the native one, as judged by the comparison of the far UV-circular dichroism spectra of rCEL-IV and native CEL-IV (nCEL-IV). Carbohydrate-binding specificity of rCEL-IV was confirmed to be similar to that of nCEL-IV from the results of the binding-inhibition assay using liposomes composed of rabbit erythrocyte lipids. Crystals of rCEL-IV were obtained in a few days by the sitting drop vapor diffusion method. These results indicate that rCEL-IV achieved essentially correct three-dimensional structure, including the carbohydrate-binding sites, and it would be very useful for further study on the carbohydrate-recognition mechanism by mutational and X-ray crystallographic analyses. PMID:16503091

  5. Amino acid sequence and carbohydrate-binding analysis of the N-acetyl-D-galactosamine-specific C-type lectin, CEL-I, from the Holothuroidea, Cucumaria echinata.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Matsuo, Noriaki; Shiba, Kouhei; Nishinohara, Shoichi; Yamasaki, Nobuyuki; Sugawara, Hajime; Aoyagi, Haruhiko

    2002-01-01

    CEL-I is one of the Ca2+-dependent lectins that has been isolated from the sea cucumber, Cucumaria echinata. This protein is composed of two identical subunits held by a single disulfide bond. The complete amino acid sequence of CEL-I was determined by sequencing the peptides produced by proteolytic fragmentation of S-pyridylethylated CEL-I. A subunit of CEL-I is composed of 140 amino acid residues. Two intrachain (Cys3-Cys14 and Cys31-Cys135) and one interchain (Cys36) disulfide bonds were also identified from an analysis of the cystine-containing peptides obtained from the intact protein. The similarity between the sequence of CEL-I and that of other C-type lectins was low, while the C-terminal region, including the putative Ca2+ and carbohydrate-binding sites, was relatively well conserved. When the carbohydrate-binding activity was examined by a solid-phase microplate assay, CEL-I showed much higher affinity for N-acetyl-D-galactosamine than for other galactose-related carbohydrates. The association constant of CEL-I for p-nitrophenyl N-acetyl-beta-D-galactosaminide (NP-GalNAc) was determined to be 2.3 x 10(4) M(-1), and the maximum number of bound NP-GalNAc was estimated to be 1.6 by an equilibrium dialysis experiment. PMID:11866098

  6. Involvement of viral envelope GP2 in Ebola virus entry into cells expressing the macrophage galactose-type C-type lectin

    Energy Technology Data Exchange (ETDEWEB)

    Usami, Katsuaki [Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033 (Japan); Matsuno, Keita; Igarashi, Manabu [Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Sapporo 001-0020 (Japan); Denda-Nagai, Kaori [Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033 (Japan); Takada, Ayato [Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Sapporo 001-0020 (Japan); Irimura, Tatsuro, E-mail: irimura@mol.f.u-tokyo.ac.jp [Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 113-0033 (Japan)

    2011-04-01

    Highlights: {yields} Ebola virus infection is mediated by binding to and fusion with the target cells. {yields} Structural feature of the viral glycoprotein determines the infectivity. {yields} Surface C-type lectin, MGL, of macrophages and dendritic cells mediate the infection. {yields} GP2, one of glycoprotein subunits, plays an essential role in MGL-mediated infection. {yields} There is a critical amino acid residue involved in high infectivity. -- Abstract: Ebola virus (EBOV) infection is initiated by the interaction of the viral surface envelope glycoprotein (GP) with the binding sites on target cells. Differences in the mortality among different species of the Ebola viruses, i.e., Zaire ebolavirus (ZEBOV) and Reston ebolavirus (REBOV), correspond to the in vitro infectivity of the pseudo-typed virus constructed with the GPs in cells expressing macrophage galactose-type calcium-type lectin (MGL/CD301). Through mutagenesis of GP2, the transmembrane-anchored subunit of GP, we found that residues 502-527 of the GP2 sequence determined the different infectivity between VSV-ZEBOV GP and -REBOV GP in MGL/CD301-expressing cells and a histidine residue at position 516 of ZEBOV GP2 appeared essential in the differential infectivity. These findings may provide a clue to clarify a molecular basis of different pathogenicity among EBOV species.

  7. Involvement of viral envelope GP2 in Ebola virus entry into cells expressing the macrophage galactose-type C-type lectin

    International Nuclear Information System (INIS)

    Highlights: → Ebola virus infection is mediated by binding to and fusion with the target cells. → Structural feature of the viral glycoprotein determines the infectivity. → Surface C-type lectin, MGL, of macrophages and dendritic cells mediate the infection. → GP2, one of glycoprotein subunits, plays an essential role in MGL-mediated infection. → There is a critical amino acid residue involved in high infectivity. -- Abstract: Ebola virus (EBOV) infection is initiated by the interaction of the viral surface envelope glycoprotein (GP) with the binding sites on target cells. Differences in the mortality among different species of the Ebola viruses, i.e., Zaire ebolavirus (ZEBOV) and Reston ebolavirus (REBOV), correspond to the in vitro infectivity of the pseudo-typed virus constructed with the GPs in cells expressing macrophage galactose-type calcium-type lectin (MGL/CD301). Through mutagenesis of GP2, the transmembrane-anchored subunit of GP, we found that residues 502-527 of the GP2 sequence determined the different infectivity between VSV-ZEBOV GP and -REBOV GP in MGL/CD301-expressing cells and a histidine residue at position 516 of ZEBOV GP2 appeared essential in the differential infectivity. These findings may provide a clue to clarify a molecular basis of different pathogenicity among EBOV species.

  8. Identification of a novel C-type lectin from the shrimp Litopenaeus vannamei and its role in defense against pathogens infection

    Institute of Scientific and Technical Information of China (English)

    LUO Zhan; ZHANG Jiquan; LI Fuhua; ZHANG Xiaojun; LIU Chengzhang; XIANG Jianhai

    2011-01-01

    Acting as one of the pattern recognition receptors (PRRs),C-type lectin is believed to mediate pathogen recognition and plays an important role in the clearance of pathogens as part of the innate immune system.In this work,a novel C-type lectin gene (named LvLecl) was cloned from the shrimp Litopenaeus vannamei.The ORF of LvLecl is 510 bp,encoding 169 amino acids.The deduced amino acid sequence contains a putative signal peptide of 19 amino acids at the N-terminal and a carbohydrate recognition domain (CRD) at the C-terminal.LvLecl was mainly expressed in the hepatopancreas.Real-time PCR analysis indicated that the level of LvLecl transcripts significantly changed in the hepatopancreas after the shrimp were artificially challenged with LPS,Micrococcus lysodeikticus and white spot syndrome virus (WSSV).RNAi-based silencing of LvLecl resulted in increases in mortality when the shrimp were challenged with WSSV,and the median lethal time was reduced compared with controls.Although there was no characteristic “EPN” (Glu-Pro-Ser) or “QPD” (Gln-Pro-Asp) motif,the recombinant LvLecl,expressed in Escherichia coli BL21 (DE3),could also agglutinate M.lysodeikticus and Vibrio anguillarum.The agglutinating activities were calcium-dependent and could be inhibited by D-mannose,D-glucose,D-galactose and N-Acetyl-D-mannose.These results suggest that LvLecl might be involved in the immune response against WSSV and bacterial infections and contribute to non-self recognition as a pattem recognition receptor in the innate immune system of the shrimp L.vannamei.

  9. CLEC4F is an inducible C-type lectin in F4/80-positive cells and is involved in alpha-galactosylceramide presentation in liver.

    Directory of Open Access Journals (Sweden)

    Chih-Ya Yang

    Full Text Available CLEC4F, a member of C-type lectin, was first purified from rat liver extract with high binding affinity to fucose, galactose (Gal, N-acetylgalactosamine (GalNAc, and un-sialylated glucosphingolipids with GalNAc or Gal terminus. However, the biological functions of CLEC4F have not been elucidated. To address this question, we examined the expression and distribution of murine CLEC4F, determined its binding specificity by glycan array, and investigated its function using CLEC4F knockout (Clec4f-/- mice. We found that CLEC4F is a heavily glycosylated membrane protein co-expressed with F4/80 on Kupffer cells. In contrast to F4/80, CLEC4F is detectable in fetal livers at embryonic day 11.5 (E11.5 but not in yolk sac, suggesting the expression of CLEC4F is induced as cells migrate from yolk cells to the liver. Even though CLEC4F is not detectable in tissues outside liver, both residential Kupffer cells and infiltrating mononuclear cells surrounding liver abscesses are CLEC4F-positive upon Listeria monocytogenes (L. monocytogenes infection. While CLEC4F has strong binding to Gal and GalNAc, terminal fucosylation inhibits CLEC4F recognition to several glycans such as Fucosyl GM1, Globo H, Bb3∼4 and other fucosyl-glycans. Moreover, CLEC4F interacts with alpha-galactosylceramide (α-GalCer in a calcium-dependent manner and participates in the presentation of α-GalCer to natural killer T (NKT cells. This suggests that CLEC4F is a C-type lectin with diverse binding specificity expressed on residential Kupffer cells and infiltrating monocytes in the liver, and may play an important role to modulate glycolipids presentation on Kupffer cells.

  10. Characterization of recombinant CEL-I, a GalNAc-specific C-type lectin, expressed in Escherichia coli using an artificial synthetic gene.

    Science.gov (United States)

    Hatakeyama, Tomomitsu; Shiba, Kouhei; Matsuo, Noriaki; Fujimoto, Tokiko; Oda, Tatsuya; Sugawara, Hajime; Aoyagi, Haruhiko

    2004-01-01

    CEL-I is a C-type lectin isolated from the Holothuroidea Cucumaria echinata. This lectin shows very high N-acetylgalactosamine-binding specificity. We constructed an artificial gene encoding recombinant CEL-I (rCEL-I) using a combination of synthetic oligonucleotides, and expressed it in Escherichia coli cells. Since the recombinant protein was obtained as inclusion bodies, the latter were solubilized using urea and 2-mercaptoethanol, and the protein was refolded during the purification and dialysis steps. The purified rCEL-I showed comparable hemagglutinating activity to that of native CEL-I at relatively high Ca(2+)-concentrations, whereas it was weaker at lower Ca(2+)-concentrations due to decreased Ca(2+)-binding affinity. rCEL-I exhibited similar carbohydrate-binding specificity to native CEL-I, including strong GalNAc-binding specificity, as examined by hemagglutination inhibition assay. Comparison of the far UV-CD spectra of recombinant and native CEL-I revealed that the two proteins undergo a similar conformational change upon binding of Ca(2+). Single crystals of rCEL-I were also obtained under the same conditions as those used for the native protein, suggesting that they have similar tertiary structures. Although native CEL-I exhibited strong cytotoxicity toward cultured cells, rCEL-I showed low cytotoxicity. These results indicate that rCEL-I has a tertiary structure and carbohydrate-binding specificity similar to those of native CEL-I. Howeger, there is a subtle difference in the properties between the two proteins probably due to the additional methionine residue at the N-terminus of rCEL-I. PMID:14999015

  11. Lectins with Potential for Anti-Cancer Therapy

    Directory of Open Access Journals (Sweden)

    Tammy Yau

    2015-02-01

    Full Text Available This article reviews lectins of animal and plant origin that induce apoptosis and autophagy of cancer cells and hence possess the potential of being developed into anticancer drugs. Apoptosis-inducing lectins encompass galectins, C-type lectins, annexins, Haliotis discus discus lectin, Polygonatum odoratum lectin, mistletoe lectin, and concanavalin A, fucose-binding Dicentrarchus labrax lectin, and Strongylocentrotus purpuratus lectin, Polygonatum odoratum lectin, and mistletoe lectin, Polygonatum odoratum lectin, autophagy inducing lectins include annexins and Polygonatum odoratum lectin.

  12. CEL-I, an N-acetylgalactosamine (GalNAc)-specific C-type lectin, induces nitric oxide production in RAW264.7 mouse macrophage cell line.

    Science.gov (United States)

    Yamanishi, Tomohiro; Hatakeyama, Tomomitsu; Yamaguchi, Kenichi; Oda, Tatsuya

    2009-08-01

    We found that CEL-I, a GalNAc-specific C-type lectin isolated from the marine invertebrate Holothuroidea (Cucumaria echinata), induces inducible nitric oxide synthase (iNOS) expression and NO production in RAW264.7 cells. The NO production was inhibited by an iNOS inhibitor, L-NAME, but was not by a lipopolysaccharide (LPS) inhibitor, polymyxin B. In the presence of 0.1-M GalNAc, increased NO production by CEL-I-treated RAW264.7 cells was observed rather than the inhibition. Bovine serum albumin (BSA) significantly inhibited the CEL-I-induced NO production as well as the binding of FITC-labelled CEL-I on RAW264.7 cells. Three MAP kinase inhibitors (specific to extra-cellular regulated kinase, c-jun NH(2)-terminal kinase and p38 MAP kinase) inhibited CEL-I-induced NO production with different extents. Heat-treatment of CEL-I resulted in a decreased activity of CEL-I depending on the temperature. These results suggest that CEL-I induces NO production in RAW264.7 cells through the protein-cell interaction rather than the binding to the specific carbohydrate chains on the cell surface. PMID:19351706

  13. Vixapatin (VP12, a C-Type Lectin-Protein from Vipera xantina palestinae Venom: Characterization as a Novel Anti-angiogenic Compound

    Directory of Open Access Journals (Sweden)

    Philip Lazarovici

    2012-10-01

    Full Text Available A C-type lectin-like protein (CTL, originally identified as VP12 and lately named Vixapatin, was isolated and characterized from Israeli viper Vipera xantina palestinae snake venom. This CTL was characterized as a selective α2β1 integrin inhibitor with anti-melanoma metastatic activity. The major aim of the present study was to prove the possibility that this protein is also a potent novel anti-angiogenic compound. Using an adhesion assay, we demonstrated that Vixapatin selectively and potently inhibited the α2 mediated adhesion of K562 over-expressing cells, with IC50 of 3 nM. 3 nM Vixapatin blocked proliferation of human dermal microvascular endothelial cells (HDMEC; 25 nM inhibited collagen I induced migration of human fibrosarcoma HT-1080 cells; and 50 nM rat C6 glioma and human breast carcinoma MDA-MB-231 cells. 1 µM Vixapatin reduced HDMEC tube formation by 75% in a Matrigel assay. Furthermore, 1 µM Vixapatin decreased by 70% bFGF-induced physiological angiogenesis, and by 94% C6 glioma-induced pathological angiogenesis, in shell-less embryonic quail chorioallantoic membrane assay. Vixapatin’s ability to inhibit all steps of the angiogenesis process suggest that it is a novel pharmacological tool for studying α2β1 integrin mediated angiogenesis and a lead compound for the development of a novel anti-angiogenic/angiostatic/anti-cancer drug.

  14. The C-type lectin receptor SIGNR3 binds to fungi present in commensal microbiota and influences immune regulation in experimental colitis

    Directory of Open Access Journals (Sweden)

    Magdalena eEriksson

    2013-07-01

    Full Text Available Inflammatory bowel disease is a condition of acute and chronic inflammation of the gut. An important factor contributing to pathogenesis is a dysregulated mucosal immunity against commensal bacteria and fungi. Host pattern recognition receptors sense commensals in the gut and are involved in maintaining the balance between controlled responses to pathogens and overwhelming innate immune activation. C-type lectin receptors (CLRs are pattern recognition receptors recognizing glycan structures on pathogens and self-antigens. Here we examined the role of the murine CLR SIGNR3 in the recognition of commensals and its involvement in intestinal immunity. SIGNR3 is the closest murine homologue of the human DC-SIGN receptor recognizing similar carbohydrate ligands such as terminal fucose or high-mannose glycans. We discovered that SIGNR3 recognizes fungi present in the commensal microbiota. To analyze if this interaction impacts the intestinal immunity against microbiota, the dextran sulfate sodium (DSS-induced colitis model was employed. SIGNR3-/- mice exhibited an increased weight loss associated with more severe colitis symptoms compared to wild-type control mice. The increased inflammation in SIGNR3-/- mice was accompanied by a higher level of TNF-α in colon. Our findings demonstrate for the first time that SIGNR3 recognizes intestinal fungi and has an immune regulatory role in colitis.

  15. Macrophage-inducible C-type lectin is associated with anti-cyclic citrullinated peptide antibodies-positive rheumatoid arthritis in men

    Institute of Scientific and Technical Information of China (English)

    WU Xin-yu; GUO Jian-ping; YIN Fang-rui; LU Xiao-lan; LI Ru; HE Jing; LIU Xu; LI Zhan-guo

    2012-01-01

    Background Macrophage-inducible C-type lectin (MINCLE) is an important member of C-type lectin superfamily,which has been shown evidence for susceptibility to arthritis in animal models.We aimed to investigate the possible association of MINCLE with rheumatoid arthritis (RA) susceptibility in Chinese Hart population.Methods Haplotypes from HapMap database (Chinese Hart Beijing,CHB) were used to select tag-single nucleotide polymorphism (SNP) (r2=0.8) residing in MINCLE gene.A total of 563 patients with RA and 404 healthy controls were TagMan genotyped for SNP rs10841845.Association analyses were performed on the whole data set and on RA subsets based on gender difference and the status of anti-cyclic citrullinated peptide (anti-CCP) antibody in RA patients.Association statistics were calculated by age and sex adjusted logistic regression.Results Overall,MINCLE SNP rs10841845 was not associated with susceptibility to RA.However,following anti-CCP stratification,rs10841845 GG genotypes conferred a significantly protective effects against anti-CCP-positive RA (OR 0.65,95% CI 0.430-0.995,P=0.048).Following gender stratification,SNP rs10841845 G allele appeared to insert its RA protective effect only in male patients,both at allele level (G vs.A OR 0.66,95% CI 0.46-0.93,P=0.018) and at genotype level (GG vs.AA+AG,OR 0.429,95% CI 0.20-0.95,P=0.036).Notably,the male RA protective effect of rs10841845 G allele was only seen in anti-CCP-positive RA (G vs.A:OR 0.64,95% CI 0.43-0.96,P=0.029; GG vs.AA+AG:OR 0.375,95% Cl 0.14-0.94,P=0.038).Furthermore,we observed a significant reduction of Disease Activity Score (DAS) 28 score (3.91±0.70 vs.5.66±0.31,P=0.022) and serum C-reactive protein levels (31.64±24.13 vs.91.80±12.02,P=0.012)in male anti-CCP-positive RA patients carrying rs10841845 GG genotype,compared with patients carrying AA+AG genotypes.Conclusions Our study provides the evidence for a gender specific association between MINCLE rs10841845 and RA

  16. Super-resolution imaging of C-type lectin spatial rearrangement within the dendritic cell plasma membrane at fungal microbe contact sites

    Directory of Open Access Journals (Sweden)

    Michelle S Itano

    2014-08-01

    Full Text Available Dendritic cells express DC-SIGN and CD206, C-type lectins (CTLs that bind a variety of pathogens and may facilitate pathogen uptake for subsequent antigen presentation. Both proteins form punctate membrane nanodomains (~80 nm on naïve cells. We analyzed the spatiotemporal distribution of CTLs following host-fungal particle contact using confocal microscopy and three distinct methods of cluster identification and measurement of receptor clusters in super-resolution datasets: DBSCAN, Pair Correlation and a custom implementation of the Getis spatial statistic. Quantitative analysis of confocal and super-resolution images demonstrated that CTL nanodomains become concentrated in the contact site relative to non-contact membrane after the first hour of exposure and established that this recruitment is sustained out to four hours. DC-SIGN nanodomains in fungal contact sites exhibit a 70% area increase and a 38% decrease in interdomain separation. Contact site CD206 nanodomains possess 90% greater area and 42% lower interdomain separation relative to non-contact regions. Contact site CTL clusters appear as disk-shaped domains of approximately 150-175 nm in diameter. The increase in length scale of CTL nanostructure in contact sites suggests that the smaller nanodomains on resting membranes may merge during fungal nanodomain structure, or that they become packed closely enough to achieve sub-resolution inter-domain edge separations of < 30 nm. This study provides evidence of local receptor spatial rearrangements on the nanoscale that occur in the plasma membrane upon pathogen binding and may direct important signaling interactions required to recognize and respond to the presence of a relatively large pathogen.

  17. Super-resolution imaging of C-type lectin spatial rearrangement within the dendritic cell plasma membrane at fungal microbe contact sites

    Science.gov (United States)

    Itano, Michelle; Graus, Matthew; Pehlke, Carolyn; Wester, Michael; Liu, Ping; Lidke, Keith; Thompson, Nancy; Jacobson, Ken; Neumann, Aaron

    2014-08-01

    Dendritic cells express DC-SIGN and CD206, C-type lectins (CTLs) that bind a variety of pathogens and may facilitate pathogen uptake for subsequent antigen presentation. Both proteins form punctate membrane nanodomains (~80 nm) on naïve cells. We analyzed the spatiotemporal distribution of CTLs following host-fungal particle contact using confocal microscopy and three distinct methods of cluster identification and measurement of receptor clusters in super-resolution datasets: DBSCAN, Pair Correlation and a custom implementation of the Getis spatial statistic. Quantitative analysis of confocal and super-resolution images demonstrated that CTL nanodomains become concentrated in the contact site relative to non-contact membrane after the first hour of exposure and established that this recruitment is sustained out to four hours. DC-SIGN nanodomains in fungal contact sites exhibit a 70% area increase and a 38% decrease in interdomain separation. Contact site CD206 nanodomains possess 90% greater area and 42% lower interdomain separation relative to non-contact regions. Contact site CTL clusters appear as disk-shaped domains of approximately 150-175 nm in diameter. The increase in length scale of CTL nanostructure in contact sites suggests that the smaller nanodomains on resting membranes may merge during fungal nanodomain structure, or that they become packed closely enough to achieve sub-resolution inter-domain edge separations of < 30 nm. This study provides evidence of local receptor spatial rearrangements on the nanoscale that occur in the plasma membrane upon pathogen binding and may direct important signaling interactions required to recognize and respond to the presence of a relatively large pathogen.

  18. Monocyte galactose/N-acetylgalactosamine-specific C-type lectin receptor stimulant immunotherapy of an experimental glioma. Part II: combination with external radiation improves survival

    International Nuclear Information System (INIS)

    A peptide mimetic of a ligand for the galactose/N-acetylgalactosamine-specific C-type lectin receptors (GCLR) exhibited monocyte-stimulating activity, but did not extend survival when applied alone against a syngeneic murine malignant glioma. In this study, the combined effect of GCLRP with radiation was investigated. C57BL/6 mice underwent stereotactic intracranial implantation of GL261 glioma cells. Animals were grouped based on randomized tumor size by magnetic resonance imaging on day seven. One group that received cranial radiation (4 Gy on days seven and nine) only were compared with animals treated with radiation and GCLRP (4 Gy on days seven and nine combined with subcutaneous injection of 1 nmol/g on alternative days beginning on day seven). Magnetic resonance imaging was used to assess tumor growth and correlated with survival rate. Blood and brain tissues were analyzed with regard to tumor and contralateral hemisphere using fluorescence-activated cell sorting analysis, histology, and enzyme-linked immunosorbent assay. GCLRP activated peripheral monocytes and was associated with increased blood precursors of dendritic cells. Mean survival increased (P < 0.001) and tumor size was smaller (P < 0.02) in the GCLRP + radiation group compared to the radiation-only group. Accumulation of dendritic cells in both the tumoral hemisphere (P < 0.005) and contralateral tumor-free hemisphere (P < 0.01) was associated with treatment. Specific populations of monocyte-derived brain cells develop critical relationships with malignant gliomas. The biological effect of GCLRP in combination with radiation may be more successful because of the damage incurred by tumor cells by radiation and the enhanced or preserved presentation of tumor cell antigens by GCLRP-activated immune cells. Monocyte-derived brain cells may be important targets for creating effective immunological modalities such as employing the receptor system described in this study

  19. Molecular characterization of the reniform nematode C-type lectin gene family reveals a likely role in mitigating environmental stresses during plant parasitism.

    Science.gov (United States)

    Ganji, Satish; Jenkins, Johnie N; Wubben, Martin J

    2014-03-10

    The reniform nematode, Rotylenchulus reniformis, is a damaging semi-endoparasitic pathogen of more than 300 plant species. Transcriptome sequencing of R. reniformis parasitic females revealed an enrichment for sequences homologous to C-type lectins (CTLs), an evolutionarily ancient family of Ca(+2)-dependent carbohydrate-binding proteins that are involved in the innate immune response. To gain further insight as to the potential role of CTLs in facilitating plant parasitism by R. reniformis, we performed a comprehensive assessment of the CTL gene family. 5'- and 3'-RACE experiments identified a total of 11 R. reniformis CTL transcripts (Rr-ctl-1 through Rr-ctl-11) that ranged in length from 1083 to 1,194 bp and showed 93-99% identity with one another. An alignment of cDNA and genomic sequences revealed three introns with the first intron residing within the 5'-untranslated region. BLAST analyses showed the closest homologs belonging to the parasitic nematodes Heligmosomoides polygyrus and Heterodera glycines. Rr-ctl-1, -2, and -3 were expressed throughout the R. reniformis life cycle; whereas, the remaining Rr-ctl genes showed life stage-specific expression. Quantitative real time RT-PCR determined that Rr-ctl transcripts were 839-fold higher in sedentary female nematodes than the next most abundant life stage. Predicted Rr-CTL peptides ranged from 301 to 338 amino acids long, possessed an N-terminal signal peptide for secretion, and contained a conserved CLECT domain, including the mannose-binding motifs EPN and EPD and the conserved WND motif that is required for binding Ca(+2). In addition, Rr-CTL peptides harbored repeats of a novel 17-mer motif within their C-terminus that showed similarity to motifs associated with bacterial ice nucleation proteins. In situ hybridization of Rr-ctl transcripts within sedentary females showed specific accumulation within the hypodermis of the body regions exposed to the soil environment; those structures embedded within the

  20. A novel C-type lectin, Nattectin-like protein, with a wide range of bacterial agglutination activity in large yellow croaker Larimichthys crocea.

    Science.gov (United States)

    Lv, Changhuan; Zhang, Dongling; Wang, Zhiyong

    2016-03-01

    C-type lectins (CTLs) are generally recognized as a superfamily of Ca(2+)-dependent carbohydrate-binding proteins, which serve as pattern recognition receptors (PRRs) in innate immunity of vertebrates. In this study, the molecular characterization and immune roles of a novel CTL from Larimichthys crocea (designated as LcNTC) were investigated. LcNTC is a novel protein that shared 33%-49% homology with other teleosts CTLs. The full-length cDNA of LcNTC was composed of 859 bp with a 465 bp open reading frame encoding a putative protein of 154 residues. LcNTC contained a single CRD with four conserved disulfide-bonded cysteine residues (Cys(57)-Cys(148), Cys(126)-Cys(140)) and EPN/AND motifs instead of invariant EPN/WND motifs required for carbohydrate-binding specificity and constructing Ca(2+)-binding sites. LcNTC mRNA was detected in all examined tissues with the most abundant in the gill. After challenged with poly I:C and Vibrio parahaemolyticus, the temporal expression of LcNTC was significantly up-regulated in the liver, spleen and head-kidney. LcNTC transcripts were also induced in the gill, skin, spleen and head-kidney post-infection with Cryptocaryon irritans. The recombinant LcNTC (rLcNTC) purified from Escherichia coli BL21 (DE3) exhibited strong agglutination activity against erythrocytes from human, rabbit and large yellow croaker in a Ca(2+)-dependent manner, and the agglutination could be inhibited by d-Mannose, d-Glucose, d-Fructose, α-Lactose, d-Maltose and LPS. Positive microbial agglutination activities of rLcNTC were observed against all tested bacteria in the presence of Ca(2+), including Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus and Micrococcus lysoleikticus) and Gram-negative bacteria (E. coli, V. parahaemolyticus, Vibrio alginolyticus and Aeromonas hydrophila). These findings collectively indicated that LcNTC might be involved in the innate immunity of L. crocea as a PRR. PMID:26828263

  1. The lipopolysaccharide-binding protein participating in hemocyte nodule formation in the silkworm Bombyx mori is a novel member of the C-type lectin superfamily with two different tandem carbohydrate-recognition domains.

    Science.gov (United States)

    Koizumi, N; Imamura, M; Kadotani, T; Yaoi, K; Iwahana, H; Sato, R

    1999-01-25

    We recently isolated and characterized the lipopolysaccharide (LPS)-binding protein, BmLBP, from the larval hemolymph of the silkworm Bombyx mori. BmLBP is a pattern recognition molecule that recognizes the lipid A portion of LPS and participates in a cellular defense reaction. This paper describes the cDNA cloning of BmLBP. The deduced amino acid sequence of BmLBP revealed that BmLBP is a novel member of the C-type lectin superfamily with a unique structural feature that consists of two different carbohydrate-recognition domains in tandem, a short and a long form. PMID:9989592

  2. Lectins with Potential for Anti-Cancer Therapy

    OpenAIRE

    Tammy Yau; Xiuli Dan; Charlene Cheuk Wing Ng; Tzi Bun Ng

    2015-01-01

    This article reviews lectins of animal and plant origin that induce apoptosis and autophagy of cancer cells and hence possess the potential of being developed into anticancer drugs. Apoptosis-inducing lectins encompass galectins, C-type lectins, annexins, Haliotis discus discus lectin, Polygonatum odoratum lectin, mistletoe lectin, and concanavalin A, fucose-binding Dicentrarchus labrax lectin, and Strongylocentrotus purpuratus lectin, Polygonatum odoratum lectin, and mistletoe lectin, Polygo...

  3. Enhancement of solubility and yield of a β-glucan receptor Dectin-1 C-type lectin-like domain in Escherichia coli with a solubility-enhancement tag.

    Science.gov (United States)

    Dulal, Hari Prasad; Nagae, Masamichi; Ikeda, Akemi; Morita-Matsumoto, Kana; Adachi, Yoshiyuki; Ohno, Naohito; Yamaguchi, Yoshiki

    2016-07-01

    Dectin-1 is a C-type lectin-like pattern recognition receptor for β(1-3)-glucans. It plays a crucial role in protecting against fungal invasion through binding to β-glucans which are commonly present on the fungal cell wall. To probe its ligand binding mechanism by NMR, we expressed the recombinant murine Dectin-1 C-type lectin-like domain (CTLD) in E. coli using pCold vector and purified it. However, the high concentration of Dectin-1 CTLD required for NMR analysis could not be attained due to its inherent low solubility and low bacterial expression. In this study, we tried to increase expression and solubility of Dectin-1 CTLD by codon optimization and fusion of a GB1 tag (B1 domain of streptococcal Protein G). GB1 was inserted on either the N-terminal (NT) or C-terminal end as well as both terminal ends of human and mouse Dectin-1 CTLDs. A pure monomeric sample was only obtained with NT-GB1 fused mouse Dectin-1. Expression of mouse Dectin-1 CTLD yielded 0.9 ± 0.2 mg/L culture, codon optimized mouse Dectin-1 CTLD produced 1.4 ± 0.2 mg/L, and the tag-fused domain 7.1 ± 0.3 mg/L. The tag also increased solubility from 0.1 mM to 1.4 mM. The recombinant protein was correctly folded, in a monomeric state, and specifically bound β-glucan laminarin. These results indicate that fusing GB1 to the N-terminus of mouse Dectin-1 domain advantageously increases yield and solubility, allows retention of native structure, and that the site of fusion is critical. PMID:27062941

  4. Structure and function of aggrecan

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Aggrecan is the major proteoglycan in the articular cartilage. This molecule is important in the proper functioning of articular cartilage because it provides a hydrated gel structure (via its interaction with hyaluronan and link protein) that endows the cartilage with load-bearing properties. It is also crucial in chondroskeletal morphogenesis during development. Aggrecan is a multimodular molecule expressed by chondrocytes. Its core protein is composed of three globular domains (Gl, G2, and G3) and a large extended region (CS) between G2 and G3 for glycosaminoglycan chain attachment. G1 comprises the amino terminus of the core protein. This domain has the same structural motif as link protein. Functionally, the G1 domain interacts with hyaluronan acid and link protein, forming stable ternary complexes in the extracellular matrix.G2 is homologous to the tandem repeats of G1 and of link protein and is involved in product processing. G3makes up the carboxyl terminus of the core protein. It enhances glycosaminoglycan modification and product secretion. Aggrecan plays an important role in mediating chondrocyte-chondrocyte and chondrocyte-matrix interactions through its ability to bind hyaluronan.

  5. Monocyte galactose/N-acetylgalactosamine-specific C-type lectin receptor stimulant immunotherapy of an experimental glioma. Part 1: stimulatory effects on blood monocytes and monocyte-derived cells of the brain

    Directory of Open Access Journals (Sweden)

    Kushchayev SV

    2012-09-01

    Full Text Available Sergiy V Kushchayev,1 Tejas Sankar,1 Laura L Eggink,4,5 Yevgeniya S Kushchayeva,5 Philip C Wiener,1,5 J Kenneth Hoober,5,6 Jennifer Eschbacher,3 Ruolan Liu,2 Fu-Dong Shi,2 Mohammed G Abdelwahab,4 Adrienne C Scheck,4 Mark C Preul11Neurosurgery Research Laboratory, 2Neuroimmunology Laboratory, 3Department of Pathology, 4Neurooncology Research, Barrow Neurological Institute, St Joseph's Hospital and Medical Center, Phoenix, 5School of Life Sciences, Arizona State University, Tempe, 6Susavion Biosciences, Inc, Tempe, AZ, USAObjectives: Immunotherapy with immunostimulants is an attractive therapy against gliomas. C-type lectin receptors specific for galactose/N-acetylgalactosamine (GCLR regulate cellular differentiation, recognition, and trafficking of monocyte-derived cells. A peptide mimetic of GCLR ligands (GCLRP was used to activate blood monocytes and populations of myeloid-derived cells against a murine glioblastoma.Methods: The ability of GCLRP to stimulate phagocytosis by human microglia and monocyte-derived cells of the brain (MDCB isolated from a human glioblastoma was initially assessed in vitro. Induction of activation markers on blood monocytes was assayed by flow cytometry after administration of GCLRP to naive mice. C57BL/6 mice underwent stereotactic intracranial implantation of GL261 glioma cells and were randomized for tumor size by magnetic resonance imaging, which was also used to assess increase in tumor size. Brain tumor tissues were analyzed using flow cytometry, histology, and enzyme-linked immunosorbent assay with respect to tumor, peritumoral area, and contralateral hemisphere regions.Results: GCLRP exhibited strong stimulatory effect on MDCBs and blood monocytes in vitro and in vivo. GCLRP was associated with an increased percentage of precursors of dendritic cells in the blood (P = 0.003, which differentiated into patrolling macrophages in tumoral (P = 0.001 and peritumoral areas (P = 0.04, rather than into dendritic cells

  6. Aggrecan structure in amphibian cartilage

    Directory of Open Access Journals (Sweden)

    Covizi D.Z.

    2000-01-01

    Full Text Available The structure of the large proteoglycan present in the bullfrog epiphyseal cartilage was studied by immunochemical and biochemical methods. The isolated monomer showed a polydisperse behavior on Sepharose CL2B, with a peak at Kav = 0.14. Chondroitin sulfate chains were identified by HPLC analysis of the products formed by chondroitinase digestion and mercuric acetate treatment. These chains have approximately 38 disaccharides, a Di45:Di68 ratio of 1.6 and GalNAc4S + GalNAc4,6S are the main non-reducing terminals. Keratan sulfate was identified by the use of two monoclonal antibodies in Western blots after chondroitinase ABC treatment. A keratan sulfate-rich region (~110 kDa was isolated by sequential treatment with chondroitinase ABC and proteases. We also employed antibodies in Western blotting experiments and showed that the full length deglycosylated core protein is about 300 kDa after SDS-PAGE. Domain-specific antibodies revealed the presence of immunoreactive sites corresponding to G1/G2 and G3 globular domains and the characterization of this large proteoglycan as aggrecan. The results indicate the high conservation of the aggrecan domain structure in this lower vertebrate.

  7. C-type lectin receptor dectin-3 mediates trehalose 6,6'-dimycolate (TDM)-induced Mincle expression through CARD9/Bcl10/MALT1-dependent nuclear factor (NF)-κB activation.

    Science.gov (United States)

    Zhao, Xue-Qiang; Zhu, Le-Le; Chang, Qing; Jiang, Changying; You, Yun; Luo, Tianming; Jia, Xin-Ming; Lin, Xin

    2014-10-24

    Previous studies indicate that both Dectin-3 (also called MCL or Clec4d) and Mincle (also called Clec4e), two C-type lectin receptors, can recognize trehalose 6,6'-dimycolate (TDM), a cell wall component from mycobacteria, and induce potent innate immune responses. Interestingly, stimulation of Dectin-3 by TDM can also induce Mincle expression, which may enhance the host innate immune system to sense Mycobacterium infection. However, the mechanism by which Dectin-3 induces Mincle expression is not fully defined. Here, we show that TDM-induced Mincle expression is dependent on Dectin-3-mediated NF-κB, but not nuclear factor of activated T-cells (NFAT), activation, and Dectin-3 induces NF-κB activation through the CARD9-BCL10-MALT1 complex. We found that bone marrow-derived macrophages from Dectin-3-deficient mice were severely defective in the induction of Mincle expression in response to TDM stimulation. This defect is correlated with the failure of TDM-induced NF-κB activation in Dectin-3-deficient bone marrow-derived macrophages. Consistently, inhibition of NF-κB, but not NFAT, impaired TDM-induced Mincle expression, whereas NF-κB, but not NFAT, binds to the Mincle promoter. Dectin-3-mediated NF-κB activation is dependent on the CARD9-Bcl10-MALT1 complex. Finally, mice deficient for Dectin-3 or CARD9 produced much less proinflammatory cytokines and keyhole limpet hemocyanin (KLH)-specific antibodies after immunization with an adjuvant containing TDM. Overall, this study provides the mechanism by which Dectin-3 induces Mincle expression in response to Mycobacterium infection, which will have significant impact to improve adjuvant and design vaccine for antimicrobial infection. PMID:25202022

  8. Monocyte galactose/N-acetylgalactosamine-specific C-type lectin receptor stimulant immunotherapy of an experimental glioma. Part 1: stimulatory effects on blood monocytes and monocyte-derived cells of the brain

    International Nuclear Information System (INIS)

    Immunotherapy with immunostimulants is an attractive therapy against gliomas. C-type lectin receptors specific for galactose/N-acetylgalactosamine (GCLR) regulate cellular differentiation, recognition, and trafficking of monocyte-derived cells. A peptide mimetic of GCLR ligands (GCLRP) was used to activate blood monocytes and populations of myeloid-derived cells against a murine glioblastoma. The ability of GCLRP to stimulate phagocytosis by human microglia and monocyte-derived cells of the brain (MDCB) isolated from a human glioblastoma was initially assessed in vitro. Induction of activation markers on blood monocytes was assayed by flow cytometry after administration of GCLRP to naive mice. C57BL/6 mice underwent stereotactic intracranial implantation of GL261 glioma cells and were randomized for tumor size by magnetic resonance imaging, which was also used to assess increase in tumor size. Brain tumor tissues were analyzed using flow cytometry, histology, and enzyme-linked immunosorbent assay with respect to tumor, peritumoral area, and contralateral hemisphere regions. GCLRP exhibited strong stimulatory effect on MDCBs and blood monocytes in vitro and in vivo. GCLRP was associated with an increased percentage of precursors of dendritic cells in the blood (P = 0.003), which differentiated into patrolling macrophages in tumoral (P = 0.001) and peritumoral areas (P = 0.04), rather than into dendritic cells, as in control animals. Treatment with GCLRP did not result in a significant change in survival of mice bearing a tumor. In vitro and in vivo activation of monocytes was achieved by administration of GCLR to mice. GCLRP-activated blood monocytes were recruited to the brain and exhibited specific phenotypes corresponding with tumor region (glioma, peritumoral zone, and contralateral glioma-free hemisphere). GCLRP treatment alone was associated with increased glioma mass as the result of the infiltration of phagocytic cells. Regional specificity for MDCB may have

  9. CEL-I, an invertebrate N-acetylgalactosamine-specific C-type lectin, induces TNF-alpha and G-CSF production by mouse macrophage cell line RAW264.7 cells.

    Science.gov (United States)

    Yamanishi, Tomohiro; Yamamoto, Yoshiko; Hatakeyama, Tomomitsu; Yamaguchi, Kenichi; Oda, Tatsuya

    2007-11-01

    Our previous studies demonstrated that CEL-I, an N-acetylgalactosamine (GalNAc)-specific C-type lectin purified from the marine invertebrate Cucumaria echinata (Holothuroidea) showed potent cytotoxicity to several cell lines such as HeLa, MDCK and XC cells. In this study, we found that CEL-I induced increased secretion of tumour necrosis factor-alpha (TNF-alpha) and granulocyte colony stimulation factor (G-CSF) by mouse macrophage cell line RAW264.7 cells in a dose-dependent manner, whereas this cell line was highly resistant to CEL-I cytotoxicity. The cytokine-inducing activity of CEL-I was stronger than that of phytohaemagglutinin (PHA-L). A binding study using FITC-labelled CEL-I (F-CEL-I) indicated that the amount of bound F-CEL-I on RAW264.7 cells was greater than that of F-PHA-L, suggesting that the greater activity of CEL-I to induce cytokine secretion by RAW264.7 cells is partly due to the higher binding ability. Since the cell binding and cytokine-inducing activity of CEL-I were partly but significantly inhibited by the specific sugar (GalNAc), it is considered that the binding of CEL-I to cell-surface-specific saccharide moieties, which may be recognized by CEL-I with higher affinity than GalNAc, is essential for the induction of cytokine secretion. The secretion of TNF-alpha and G-CSF from CEL-I-treated RAW264.7 cells were almost completely prevented by brefeldin A (BFA), whereas increase in mRNA levels of these cytokines were not affected by BFA. Bio-Plex beads assay suggested that temporal increase in phosphorylation of extracellular-regulated kinase (ERK), c-jun NH(2)-terminal kinase (JNK) and p38 MAP kinase occurred at relatively early time following CEL-I treatment. Furthermore, the secretion of TNF-alpha and G-CSF were inhibited by specific inhibitors for these MAP kinases. These results suggest that the intracellular signal transduction through the activation of MAP kinase system is involved in CEL-I-induced cytokine secretion. PMID:17846063

  10. Cartilage Aggrecan Can Undergo Self-Adhesion

    OpenAIRE

    Han, Lin; Dean, Delphine; Daher, Laura A.; Grodzinsky, Alan J.; Ortiz, Christine

    2008-01-01

    Here it is reported that aggrecan, the highly negatively charged macromolecule in the cartilage extracellular matrix, undergoes Ca2+-mediated self-adhesion after static compression even in the presence of strong electrostatic repulsion in physiological-like solution conditions. Aggrecan was chemically end-attached onto gold-coated planar silicon substrates and gold-coated microspherical atomic force microscope probe tips (end radius R ≈ 2.5 μm) at a density (∼40 mg/mL) that simulates physiolo...

  11. Expression of C-type lectin receptors and Toll-like receptors in decidua of unexplained recurrent spontaneous abortion. Expression of C-type lectin receptors and Toll-like receptors in decidua of unexplained recurrent spontaneous abortion%C 型凝集素受体和 Toll 样受体在原因不明复发性流产患者蜕膜中的表达

    Institute of Scientific and Technical Information of China (English)

    邱添; 滕银成; 王玉东; 徐亮

    2014-01-01

    目的:研究原因不明复发性流产(URSA)患者蜕膜中树突细胞(DC)特异性细胞间黏附分子-3结合非整合素因子(DC-SIGN)和甘露糖受体( MR)及 Toll 样受体2(TLR-2)和 TLR-4的表达情况。方法:采集 URSA 患者及正常妊娠人工流产妇女(对照组)的新鲜蜕膜。采用免疫组化和 Western blot 法检测蜕膜中 DC-SIGN、MR、TLR-2和TLR-4蛋白表达,RT-PCR 检测蜕膜中 DC-SIGN、MR、TLR-2和 TLR-4 mRNA 表达;流式细胞检测蜕膜中 DC 表面表达 DC-SIGN、MR、TLR-2和 TLR-4的水平。结果:与对照组比较, URSA 组蜕膜中及 DC 表面的 TLR-2和 TLR-4表达水平明显升高(P0.05)。结论:相对于正常妊娠妇女,URSA 患者蜕膜中及树突细胞表面的 C 型凝集素受体(CLRs)弱势表达,而 TLRs 优势表达。 CLRs 与 TLRs 之间的免疫平衡失调可能是 UR-SA 发病机制中的重要因素之一。%Objective:To investigate the expression of C-typed lectin receptors (CLRs) of DC-SIGN and MR and Toll-like receptors (TLRs) of TLR-2 and TLR-4 in decidua of unex-plained recurrent spontaneous abortion( URSA) patients. Methods:The decidua tissues were obtained from URSA patients (URSA group,n =20) and normal pregnant women (NP group,n= 20). The expressions of DCSIGN,MR,TLR-2 and TLR-4 protein in decidua were detected by immunohistochemistry and Western blot. The levels of DCSIGN,MR,TLR-2 and TLR-4 mRNA in decidua were detected by RT-PCR. The expressions of DCSIGN,MR,TLR-2 and TLR-4 mol-ecule on DC were detected by flow cytometry. Results:The expressions of TLR-2 and TLR-4 on DC and in decidua in URSA group were higher than that in NP group (P0. 05). Conclusion:Compared with normal pregnant women,the weak expression of CLRs and the overexpression of TLRs were observed in decidua in URSA patients. The imbal-ance between CLRs and TLRs may play an important role in the pathogenesis of URSA.

  12. The different roles of aggrecan interaction domains

    DEFF Research Database (Denmark)

    Aspberg, Anders

    2012-01-01

    vital in that it binds the proteoglycan to hyaluronan in ternary complex with link protein, retaining the proteoglycan in the tissue. The importance of the C-terminal G3 domain interactions has recently been emphasized by two different human hereditary disorders: autosomal recessive aggrecan...

  13. 应用3'RACE技术扩增仿刺参C型凝集素基因及实验条件的优化%Clone of C-type lectin gene of sea cucumber Apostichopus japonicus and optimization of experimental conditions using 3' RACE technique

    Institute of Scientific and Technical Information of China (English)

    李莹; 韩璐璐

    2012-01-01

    The express sequence tag (EST) of C-type leetin of Apostichopus japonicus from GenBank was obtained. Based on this sequence, we utilize software of primer 5. 0 to design a pair of RACE primer. After the PCR reaction, the cDNA sequence of C-type leetin of Apostichopus japonicus was obtained. Based on part of the known partial cDNA sequence,gene specific primers (GSPF) was designed which work together with universal primers (UPM). The 3' cDNA end of C-type leetin in Apostichopus japonicus was successfully cloned. At the same time, we optimized this experiment and an amplified fragment of 670 bp in length which overlapped the known C-type leetin sequence by 417 bp was obtained subsequently. The result showed that this sequence was consistent with expected aim gene.%从GenBank上调取刺参C型凝集素(C-type Lectin)基因序列EST,根据此序列设计RACE引物,采用PCR扩增技术得到了仿刺参C-type Lectin基因序列(EST),根据这段EST序列设计1个基因特异引物(GSPF),与通用引物(UPM)扩增,成功地克隆到了该基因的3’末端序列.同时,对仿刺参C型凝集素基因3’克隆的实验条件进行了优化.该扩增片段长度为670 bp,与已知序列重叠部分为417 bp.经测序和比对发现该段序列与预期的目标基因的序列一致.

  14. Lectin binders

    International Nuclear Information System (INIS)

    Lectins are widely distributed in the plant kingdom, many of them being well characterized in their chemical structure and the effects they have on alien biological systems such as erythrocytes or lymphocytes. The biological function of plant lectins remains speculative. We therefore inspected plant extracts from components which might bind specifically to the lectin from the respective plant. Single proteins (lectin binders) could be isolated from each plant extract. The interaction of these proteins with lectins was demonstrated and qualified by several methods. Similar to the lectins, the lectin binders are localized in the cytoplasm in contrast to them, however, they persist during germination and plant growth. Their precise role in the plant is not known, but they are likely to be associated with lectins not only in vitro but also in vivo. They also interact with alien cells, and are able to stimulate mitosis in murine lymphocytes. Some lectin binders act specifically on B lymphocytes, leaving T cells uninfluenced

  15. Patients with rheumatoid arthritis have an altered circulatory aggrecan profile

    Directory of Open Access Journals (Sweden)

    Mueller Andreas

    2008-05-01

    Full Text Available Abstract Background Rheumatoid arthritis (RA is a chronic auto-immune disease with extensive articular cartilage destruction. Aggrecan depletion, mediated by aggrecanases is one of the first signs of early cartilage erosion. We investigated, whether measurement of aggrecan and fragments thereof in serum, could be used as biomarkers for joint-disease in RA patients and furthermore characterized the fragments found in the circulation. Methods The study consisted of 38 patients, 12 males (62.2 ± 16.0 years and 26 females (59.8 ± 20.7 years diagnosed with RA: 41.5 ± 27.5 mm/h erythrocyte sedimentation rate (ESR, 38.4 ± 34.7 mg/ml C-reactive protein (CRP and 4.8 ± 1.7 disease activity score (DAS and 108 healthy age-matched controls. Aggrecan levels were measured using two immunoassays, i.e. the 374ARGSVI-G2 sandwich ELISA measuring aggrecanase-mediated aggrecan degradation and the G1/G2 sandwich assay, detecting aggrecan molecules containing G1 and/or G2 (total aggrecan We further characterized serum samples by western blots, by using monoclonal antibodies F-78, binding to G1 and G2, or by BC-3, detecting the aggrecanase-generated N-terminal 374ARGSVI neo-epitope. Results Total aggrecan levels in RA patients were significantly decreased from 824.8 ± 31 ng/ml in healthy controls to 570.5 ± 30 ng/ml (31% decrease, P Conclusion This is the first study, which characterizes different aggrecan fragments in human serum. The data strongly suggests that total aggrecan levels, i.e. aggrecan molecules containing G1 and/or G2 are lower in RA patients, and that RA patients have at least one specific subpopulation of aggrecan fragments, namely aggrecanse generated 374ARGSVI fragments. Further clinical studies are needed to investigate the potential of G1/G2 as a structure-related biochemical marker in destructive joint-diseases.

  16. A nuclear magnetic resonance study of water in aggrecan solutions

    Science.gov (United States)

    Foster, Richard J.; Damion, Robin A.; Baboolal, Thomas G.; Smye, Stephen W.; Ries, Michael E.

    2016-01-01

    Aggrecan, a highly charged macromolecule found in articular cartilage, was investigated in aqueous salt solutions with proton nuclear magnetic resonance. The longitudinal and transverse relaxation rates were determined at two different field strengths, 9.4 T and 0.5 T, for a range of temperatures and aggrecan concentrations. The diffusion coefficients of the water molecules were also measured as a function of temperature and aggrecan concentration, using a pulsed field gradient technique at 9.4 T. Assuming an Arrhenius relationship, the activation energies for the various relaxation processes and the translational motion of the water molecules were determined from temperature dependencies as a function of aggrecan concentration in the range 0–5.3% w/w. The longitudinal relaxation rate and inverse diffusion coefficient were approximately equally dependent on concentration and only increased by upto 20% from that of the salt solution. The transverse relaxation rate at high field demonstrated greatest concentration dependence, changing by an order of magnitude across the concentration range examined. We attribute this primarily to chemical exchange. Activation energies appeared to be approximately independent of aggrecan concentration, except for that of the low-field transverse relaxation rate, which decreased with concentration. PMID:27069663

  17. C型凝集素受体和Toll样受体在子(癎)前期重度患者胎盘中的表达%Expression of C-type lectin receptors and Toil-like receptors in placenta of severe pre-eclampsia

    Institute of Scientific and Technical Information of China (English)

    李黎; 徐亮; 滕银成

    2011-01-01

    Objective To investigate the expression of C-typed lectin receptors of dendritic cells specific intercellular adhesion raolecule-3-grabbing nonintegrin ( DC-SIGN) and mannose receptor (MR) and Toll-like receptors (TLR) of TLR-2 and TLR-4 in placenta tissues in patients with pre-eclampsia ( PE). Methods The placenta tissues were obtained from patients with severe pre-eclampsia ( PE group, n = 20) and normal pregnant women ( NP group, n - 20). The expression of DC-SIGN, MR, TLR-2 and TLR-4 in placenta tissues was localized by immunohistochemistry, and the expression of DC-SIGN, MR, TLR-2 and TLR-4 protein in placenta tissues was detected by Western blotting. Results There was expression of DC-SIGN, MR, TLR-2 and TLR-4 in syncytiotrophoblasts and cytotrophoblasts of placenta tissues in both groups, and the majority was in syncytiotrophoblasts. The expression of TLR-4 in placenta tissues in PE group was significantly higher than that in NP group (P 0. 05). Conclusion Compared with normal pregnant women, the expression of C-typed lectin receptors of DC-SIGN and MR is lower, while the expression of TLR-4 is abnormally higher in placenta tissues in patients with PE. The imbalance between these two receptors may play an important role in the pathogenesis of PE.%目的 分析子(癎)前期(PE)患者胎盘组织中C型凝集素样受体家族中DC-SIGN和甘露糖受体(MR)以及Toll样受体(TLR)家族中TLR-2和TLR-4的表达情况.方法 采集PE重度患者(PE组,n=20)及正常妊娠妇女(NP组,n=20)的新鲜胎盘标本;采用免疫组织化学技术对胎盘组织中DC-SIGN、MR、TLR-2和TLR-4的表达进行定位分析;采用Western blotting法检测胎盘组织中的DC-SIGN、MR、TLR-2和TLR-4的蛋白表达.结果 两组胎盘组织中均有DC-SIGN、MR、TLR-2和TLR-4的表达,主要位于胎盘合体滋养层细胞,在细胞滋养层细胞中亦有少量表达.PE组与NP组比较,胎盘组织TLR-4阳性细胞表达率明显升高(P <0.01);DC-S1GN、MR

  18. An ARGS-aggrecan assay for analysis in blood and synovial fluid

    DEFF Research Database (Denmark)

    Larsson, S; Lohmander, Stefan; Struglics, A

    2014-01-01

    the Meso Scale Discovery (MSD) platform for detection of ARGS-aggrecan was validated, using a standard made from recombinant human aggrecan. Matched samples of SF, serum, plasma, and urine were obtained from 36 subjects at different time points after knee injury, and analysed for ARGS-aggrecan content...

  19. The Development and Activity-Dependent Expression of Aggrecan in the Cat Visual Cortex

    OpenAIRE

    Kind, P. C.; Sengpiel, F.; Beaver, C. J.; Crocker-Buque, A.; Kelly, G. M.; R. T. Matthews; Mitchell, D E

    2012-01-01

    The Cat-301 monoclonal antibody identifies aggrecan, a chondroitin sulfate proteoglycan in the cat visual cortex and dorsal lateral geniculate nucleus (dLGN). During development, aggrecan expression increases in the dLGN with a time course that matches the decline in plasticity. Moreover, examination of tissue from selectively visually deprived cats shows that expression is activity dependent, suggesting a role for aggrecan in the termination of the sensitive period. Here, we demonstrate for ...

  20. MMPs are less efficient than ADAMTS5 in cleaving aggrecan core protein

    OpenAIRE

    Durigova, Michaela; NAGASE, Hideaki; Mort, John S.; Roughley, Peter J.

    2010-01-01

    Aggrecan degradation in articular cartilage occurs predominantly through proteolysis and has been attributed to the action of members of the matrix metalloproteinase (MMP) and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) families. Both families of enzymes cleave aggrecan at specific sites within the aggrecan core protein. One cleavage site within the interglobular domain (IGD), between Glu373–374Ala and five additional sites in the chondroitin sulfate-2 (CS-2) regio...

  1. Lectins from Edible Mushrooms

    Directory of Open Access Journals (Sweden)

    Senjam Sunil Singh

    2014-12-01

    Full Text Available Mushrooms are famous for their nutritional and medicinal values and also for the diversity of bioactive compounds they contain including lectins. The present review is an attempt to summarize and discuss data available on molecular weights, structures, biological properties, N-terminal sequences and possible applications of lectins from edible mushrooms. It further aims to update and discuss/examine the recent advancements in the study of these lectins regarding their structures, functions, and exploitable properties. A detailed tabling of all the available data for N-terminal sequences of these lectins is also presented here.

  2. Clinical correlations of aggrecan in the resected medial rectus muscle of patients with intermittent exotropia.

    OpenAIRE

    Yamane T; Matsuo T; Hasebe S; Ohtsuki H

    2003-01-01

    The purpose of this study was to elucidate the role of extracellular matrix components such as aggrecan, fibronectin, and laminin in the extraocular muscle of patients with strabismus. Resected tissues of the medial rectus muscle of 47 patients with intermittent exotropia obtained during recession-resection surgery were frozen under liquid nitrogen and pulverized by a Freezer/Mill to solubilize the tissue for enzyme immunoassay. The total amounts of aggrecan, fibronectin, and laminin in the r...

  3. Aggrecan nanoscale solid-fluid interactions are a primary determinant of cartilage dynamic mechanical properties.

    Science.gov (United States)

    Nia, Hadi Tavakoli; Han, Lin; Bozchalooi, Iman Soltani; Roughley, Peter; Youcef-Toumi, Kamal; Grodzinsky, Alan J; Ortiz, Christine

    2015-03-24

    Poroelastic interactions between interstitial fluid and the extracellular matrix of connective tissues are critical to biological and pathophysiological functions involving solute transport, energy dissipation, self-stiffening and lubrication. However, the molecular origins of poroelasticity at the nanoscale are largely unknown. Here, the broad-spectrum dynamic nanomechanical behavior of cartilage aggrecan monolayer is revealed for the first time, including the equilibrium and instantaneous moduli and the peak in the phase angle of the complex modulus. By performing a length scale study and comparing the experimental results to theoretical predictions, we confirm that the mechanism underlying the observed dynamic nanomechanics is due to solid-fluid interactions (poroelasticity) at the molecular scale. Utilizing finite element modeling, the molecular-scale hydraulic permeability of the aggrecan assembly was quantified (kaggrecan = (4.8 ± 2.8) × 10(-15) m(4)/N·s) and found to be similar to the nanoscale hydraulic permeability of intact normal cartilage tissue but much lower than that of early diseased tissue. The mechanisms underlying aggrecan poroelasticity were further investigated by altering electrostatic interactions between the molecule's constituent glycosaminoglycan chains: electrostatic interactions dominated steric interactions in governing molecular behavior. While the hydraulic permeability of aggrecan layers does not change across species and age, aggrecan from adult human cartilage is stiffer than the aggrecan from newborn human tissue. PMID:25758717

  4. Lectin typing of Campylobacter concisus

    DEFF Research Database (Denmark)

    Aabenhus, Rune Munck; Hynes, Sean O; Permin, Henrik;

    2002-01-01

    A total of 44 clinical isolates and the type strain of the putative pathogen Campylobacter concisus were grouped based on their reactions with plant lectins. The optimized lectin typing system used C. concisus strains proteolytically pretreated and subsequently typed by using a panel of four...... lectins. The system grouped all 45 strains into 13 lectin reaction patterns, leaving no strain untypeable due to autoagglutination. Lectin types were both stable and reproducible....

  5. Serum levels of mannan-binding lectin in chickens prior to and during experimental infection with avian infectious bronchitis virus

    DEFF Research Database (Denmark)

    Juul-Madsen, H.R.; Munch, M.; Handberg, Kurt; Sørensen, P.; Johnson, A.A.; Norup, L.R.; Jørgensen, Poul Henrik

    2003-01-01

    Mannan-binding lectin (MBL) is a glycoprotein and a member of the C-type lectin super family, the collectin family, and the acute phase protein family. The MBL exerts its function by directly binding to microbial surfaces through its carbohydrate recognition domains, followed by direct opsonization...

  6. Short aggrecan gene repetitive alleles associated with lumbar degenerative disc disease in Turkish patients.

    Science.gov (United States)

    Eser, O; Eser, B; Cosar, M; Erdogan, M O; Aslan, A; Yıldız, H; Solak, M; Haktanır, A

    2011-01-01

    We investigated a possible association between aggrecan gene polymorphism and lumbar degenerative disc disease in Turkish patients. One hundred 20-30-year-old patients with or without low back pain were selected for the study. Lumbar magnetic resonance imaging was performed on all patients. The patient group had low back pain clinically and degenerative disc disease radiographically. The control group included patients with and without low back pain: all were negative radiographically for degenerative disc disease. Genomic DNA was extracted from all participants. A PCR assay were used to evaluate variable number of tandem repeat polymorphism of aggrecan gene alleles to determine if there was any correlation with degenerative disc disease. Significant associations were found between short repeated alleles of the aggrecan gene and severe disc degeneration. A significant association was also found between short repeated alleles of the aggrecan gene and multilevel disc herniation as well as extrusion and sequestration types of disc herniation. In Turkish population, short repeated alleles of the aggrecan gene are associated with increased disc degeneration and disc herniation. PMID:21948754

  7. Clinical correlations of aggrecan in the resected medial rectus muscle of patients with intermittent exotropia.

    Directory of Open Access Journals (Sweden)

    Yamane T

    2003-08-01

    Full Text Available The purpose of this study was to elucidate the role of extracellular matrix components such as aggrecan, fibronectin, and laminin in the extraocular muscle of patients with strabismus. Resected tissues of the medial rectus muscle of 47 patients with intermittent exotropia obtained during recession-resection surgery were frozen under liquid nitrogen and pulverized by a Freezer/Mill to solubilize the tissue for enzyme immunoassay. The total amounts of aggrecan, fibronectin, and laminin in the resected tissue were correlated with clinical data of patients such as age, exodeviation, and refractive error. The amount of aggrecan decreased significantly with the advance of age (P < 0.0001, Spearman rank correlation test, while the amount of laminin or fibronectin had no correlation with age. Patients with basic type intermittent exotropia showed larger, although not significantly, amounts of aggrecan than those with convergence insufficiency type (P = 0.0538, Mann-Whitney U-test. The amount of aggrecan may be related to motor aspects of intermittent exotropia.

  8. Lectins with Anti-HIV Activity: A Review

    OpenAIRE

    Ouafae Akkouh; Tzi Bun Ng; Senjam Sunil Singh; Cuiming Yin; Xiuli Dan; Yau Sang Chan; Wenliang Pan; Randy Chi Fai Cheung

    2015-01-01

    Lectins including flowering plant lectins, algal lectins, cyanobacterial lectins, actinomycete lectin, worm lectins, and the nonpeptidic lectin mimics pradimicins and benanomicins, exhibit anti-HIV activity. The anti-HIV plant lectins include Artocarpus heterophyllus (jacalin) lectin, concanavalin A, Galanthus nivalis (snowdrop) agglutinin-related lectins, Musa acuminata (banana) lectin, Myrianthus holstii lectin, Narcissus pseudonarcissus lectin, and Urtica diocia agglutinin. The anti-HIV al...

  9. Involvement of ADAMTS5 and hyaluronidase in aggrecan degradation and release from OSM-stimulated cartilage

    Directory of Open Access Journals (Sweden)

    M Durigova

    2011-01-01

    Full Text Available The relative contribution of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS4 and ADAMTS5 to aggrecan degradation under oncostatin M (OSM stimulation, the role of the ancillary domains of the aggrecanases on their ability to cleave within the chondroitin sulfate (CS-2 region, the role of hyaluronidases (HYAL in stimulating aggrecan release in the absence of proteolysis, and the identity of the hyaluronidase involved in OSM-mediated cartilage breakdown were investigated. Bovine articular cartilage explants were cultured in the presence of interleukin-1beta (IL-1beta, tumor necrosis factor alpha (TNFalpha and/or OSM, or treated with trypsin and/or hyaluronidase. Aggrecan was digested with various domain-truncated isoforms of ADAMTS4 and ADAMTS5. Aggrecan and link protein degradation and release were analyzed by immunoblotting. Aggrecanase and HYAL gene expression were determined. ADAMTS4 was the most inducible aggrecanase upon cytokine stimulation, whereas ADAMTS5 was the most abundant aggrecanase. ADAMTS5 was the most active aggrecanase and was responsible for the generation of an OSM-specific degradation pattern in the CS-2 region. Its ability to cleave at the OSM-specific site adjacent to the aggrecan G3 region was enhanced by truncation of the C-terminal thrombospondin domain, but reduced by further truncation of both the spacer and cysteine-rich domains of the enzyme. OSM has the ability to mediate proteoglycan release through hyaluronan degradation, under conditions where HYAL-2 is the predominant hyaluronidase being expressed. Compared to other catabolic cytokines, OSM exhibits a unique potential at degrading the proteoglycan aggregate, by promoting early robust aggrecanolysis, primarily through the action of ADAMTS5, and hyaluronan degradation.

  10. Banana Lectin: A Brief Review

    Directory of Open Access Journals (Sweden)

    Senjam Sunil Singh

    2014-11-01

    Full Text Available Lectins are a group of proteins of non-immune origin that recognize and bind to carbohydrates without modifying them. Banana is the common name for both herbaceous plants of the genus Musa and for the fruit they produce. They are indeed a promising source for many medicinal applications. Banana lectins have the potential for inhibiting HIV-1 reverse transcriptase activity, suppressing cancer cell proliferation and stimulating macrophage activities. Nevertheless, compared to other plant lectins, there is relatively little information in the literature on banana lectins, particularly with respect to their structure and biological functions. Herein we focus our review on the structure, functions and exploitable properties of banana lectins.

  11. Nkrp1 Family, from Lectins to Protein Interacting Molecules

    Czech Academy of Sciences Publication Activity Database

    Rozbeský, Daniel; Ivanova, Ljubina; Hernychová, Lucie; Grobárová, Valeria; Novák, Petr; Černý, J.

    2015-01-01

    Roč. 20, č. 2 (2015), s. 3463-3478. ISSN 1420-3049 R&D Projects: GA MŠk(CZ) EE2.3.20.0055; GA MŠk(CZ) EE2.3.30.0003; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : NATURAL-KILLER-CELL * C-TYPE LECTIN * CARBOHYDRATE-RECOGNITION DOMAIN Subject RIV: CC - Organic Chemistry Impact factor: 2.416, year: 2014

  12. Role of calcium signaling in down-regulation of aggrecan induced by cyclic tensile strain in annulus fibrosus cells

    Institute of Scientific and Technical Information of China (English)

    GUO Zhi-liang; ZHOU Yue; LI Hua-zhuang; CAO Guo-yong; TENG Hai-jun

    2006-01-01

    Objective:To study the role of intracellular calcium signal pathway in the down-regulation of aggrecan induced by cyclic tensile strain in the annulus fibrosus cells. Methods :The expression of aggrecan mRNA and core protein were respectively detected with RT-PCR and western blot after the channels transmitting calcium ions were blocked with EGTA, gadolinium and verapamil. Results:EGTA, gadolinium and verapamil partially prevented the effects of cyclic tensile strain on the expression of aggrecan in annulus fibrosus cells. Conclusion:The calcium signaling is involved in the down-regulation of proteoglycan resulting from cyclic tensile strain in the annulus fibrosus cells.

  13. C-type lectin receptors on dendritic cells and Langerhans cells.

    NARCIS (Netherlands)

    Figdor, C.G.; Kooyk, Y. van; Adema, G.J.

    2002-01-01

    Dendritic cells and Langerhans cells are specialized for the recognition of pathogens and have a pivotal role in the control of immunity. As guardians of the immune system, they are present in essentially every organ and tissue, where they operate at the interface of innate and acquired immunity. Re

  14. A C-Type Lectin from Bothrops jararacussu Venom Disrupts Staphylococcal Biofilms

    OpenAIRE

    Raphael Contelli Klein; Mary Hellen Fabres-Klein; Leandro Licursi de Oliveira; Renato Neves Feio; François Malouin; Andréa de Oliveira Barros Ribon

    2015-01-01

    Bovine mastitis is a major threat to animal health and the dairy industry. Staphylococcus aureus is a contagious pathogen that is usually associated with persistent intramammary infections, and biofilm formation is a relevant aspect of the outcome of these infections. Several biological activities have been described for snake venoms, which led us to screen secretions of Bothrops jararacussu for antibiofilm activity against S. aureus NRS155. Crude venom was fractionated by size-exclusion chro...

  15. Biochemical and ultrastructural studies of the C-type lectin bovine conglutinin

    DEFF Research Database (Denmark)

    Andersen, Ove; Nielsen, E H; Storgaard, P;

    1992-01-01

    The aim of this study was to correlate the supramolecular organization of conglutinin (BK) with its primary and tertiary structure and to gain more knowledge of functionally important regions of the molecule. BK analyzed by SDS-PAGE under standard reducing conditions (40 mM DTT) showed a major ba...

  16. The non-aggregated aggrecan in the human intervertebral disc can arise by a non-proteolytic mechanism

    Directory of Open Access Journals (Sweden)

    PJ Roughley

    2014-09-01

    Full Text Available Analysis of both the aggregated and non-aggregated fractions of aggrecan isolated from adult human intervertebral disc using immunoblotting with antibodies specific for the different domains constituting the aggrecan core protein or atomic force microscopy revealed that many components contained the G1 domain. However, little of the disc aggrecan was able to reform aggregates with hyaluronan, as determined by gel filtration chromatography, suggesting that the G1 domains had been rendered non-functional. Since previous studies have shown that disc aggrecan undergoes non-enzymatic glycation with age, the functional effect of such modification was investigated in vitro using bovine aggrecan isolated from young animals. Incubation of monomeric aggrecan with ribose to induce glycation rendered it unable to form complexes with hyaluronan stable to agarose gel electrophoresis or gel filtration chromatography. Similarly, extended treatment of intact proteoglycan aggregate with ribose resulted in destabilisation of the complex with separation of the aggrecan from the hyaluronan. Although it is clear that proteolysis occurs in the intervertebral disc and gives rise to some non-aggregating molecules, a different mechanism is required to explain the presence of many non-aggregating molecules bearing the G1 domain. The products of non-enzymatic glycation of the globular domains of aggrecan would account for this phenomenon and explain why some of the non-aggregating molecules are still large proteoglycans. While such molecules may be retained in the nucleus pulposus, they may be able to diffuse within it, reducing the ability of the tissue to resist compression under asymmetric loading such as bending and ultimately contributing to disc degeneration.

  17. MMBL proteins: from lectin to bacteriocin.

    Science.gov (United States)

    Ghequire, Maarten G K; Loris, Remy; De Mot, René

    2012-12-01

    Arguably, bacteriocins deployed in warfare among related bacteria are among the most diverse proteinacous compounds with respect to structure and mode of action. Identification of the first prokaryotic member of the so-called MMBLs (monocot mannose-binding lectins) or GNA (Galanthus nivalis agglutinin) lectin family and discovery of its genus-specific killer activity in the Gram-negative bacteria Pseudomonas and Xanthomonas has added yet another kind of toxin to this group of allelopathic molecules. This novel feature is reminiscent of the protective function, on the basis of antifungal, insecticidal, nematicidal or antiviral activity, assigned to or proposed for several of the eukaryotic MMBL proteins that are ubiquitously distributed among monocot plants, but also occur in some other plants, fish, sponges, amoebae and fungi. Direct bactericidal activity can also be effected by a C-type lectin, but this is a mammalian protein that limits mucosal colonization by Gram-positive bacteria. The presence of two divergent MMBL domains in the novel bacteriocins raises questions about task distribution between modules and the possible role of carbohydrate binding in the specificity of target strain recognition and killing. Notably, bacteriocin activity was also demonstrated for a hybrid MMBL protein with an accessory protease-like domain. This association with one or more additional modules, often with predicted peptide-hydrolysing or -binding activity, suggests that additional bacteriotoxic proteins may be found among the diverse chimaeric MMBL proteins encoded in prokaryotic genomes. A phylogenetic survey of the bacterial MMBL modules reveals a mosaic pattern of strongly diverged sequences, mainly occurring in soil-dwelling and rhizosphere bacteria, which may reflect a trans-kingdom acquisition of the ancestral genes. PMID:23176516

  18. The lectin pathway of complement

    DEFF Research Database (Denmark)

    Ballegaard, Vibe Cecilie Diederich; Haugaard, Anna Karen; Garred, P;

    2014-01-01

    The pattern recognition molecules of the lectin complement pathway are important components of the innate immune system with known functions in host-virus interactions. This paper summarizes current knowledge of how these intriguing molecules, including mannose-binding lectin (MBL), Ficolin-1, -2...

  19. Regulation of aggrecanases from the ADAMTS family and aggrecan neoepitope formation during in vitro chondrogenesis of human mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    W Richter

    2012-05-01

    Full Text Available Aggrecanases from the ADAMTS (A Disintegrin And Metalloproteinase with ThromboSpondin motifs family are important therapeutic targets due to their essential role in aggrecan depletion in arthritic diseases. Whether their function is also important for matrix rearrangements during chondrogenesis and thus, cartilage regeneration, is however so far unknown. The aim of this study was to analyse the expression and function of ADAMTS with aggrecanase activity during chondrogenic differentiation of human mesenchymal stem cells (MSCs. Chondrogenic differentiation was induced in bone marrow-derived MSC pellets and expression of COL2A1, aggrecan, ADAMTS1, 4, 5, 9, 16 and furin was followed by quantitative RT-PCR. Formation of the NITEGE (ADAMTS-cleaved and DIPEN (MMP-cleaved aggrecan neoepitopes was detected by immunohistochemistry. While the expression of ADAMTS4, 9, 16 and furin was up-regulated during chondrogenesis, ADAMTS1 and 5 were down-regulated. Despite this regulation of ADAMTS, no formation of NITEGE neoepitopes occurred in MSC pellets, indicating no ADAMTS-induced cleavage of aggrecan. In contrast, MMP-induced cleavage of aggrecan appeared at 14 d after induction of chondrogenesis. Submission of differentiated MSC pellets to IL1β treatment for 3 d resulted in strong upregulation of ADAMTS1, 4 and 5, rapid proteoglycan depletion, and stimulation of ADAMTS-induced but not MMP-induced cleavage of aggrecan. Thus, there is no evidence for ADAMTS-induced aggrecan cleavage during chondrogenesis, but proteoglycan turnover is rapidly inducible under inflammatory signals. Therapeutic aggrecanase inhibition for treatment of arthritic disease may thus not impede regenerative self-healing pathways based on chondrogenesis of local progenitor cells in the joint.

  20. Study of serum aggrecan fragments in handigodu disease precipitated by DMB dye

    Directory of Open Access Journals (Sweden)

    Mallikarjun Badadani

    2012-04-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 Handigodu disease is a disorder of osteoarticular system, specific to particular region viz shimoga and chickmagalore districts of Karnataka state, India. The present study focuses to understand the metabolism of aggrecan, a key glycoprotein of cartilage matrix in Handigodu Disease (HD using serum specimen. The Dimethylmethylene Blue Chloride (DMB dye was used to precipitate glycosaminoglycans (GAGs associated proteins in serum. DMB dye was removed from GAGs associated proteins and electrophoresed on SDS-PAGE. The five GAGs associated protein bands were separated, among them two bands; 21.5 KDa and 55.4 KDa were immunoreactive to anti-human aggrecan antibody. These two GAGs associated protein bands were more dense (86% for 21.5 KDa and 61 % for 55.4 KDa in affected group when compared to unaffected group in Handigodu Disease. These results may imply defective metabolism of aggrecan in a disease condition. The study needs to be extend for other arthritis / dysplastic bone disorders. st1\\:*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;}

  1. Evidence for lysosomal exocytosis and release of aggrecan-degrading hydrolases from hypertrophic chondrocytes, in vitro and in vivo

    Directory of Open Access Journals (Sweden)

    Edward R. Bastow

    2012-02-01

    The abundant proteoglycan, aggrecan, is resorbed from growth plate cartilage during endochondral bone ossification, yet mice with genetically-ablated aggrecan-degrading activity have no defects in bone formation. To account for this apparent anomaly, we propose that lysosomal hydrolases degrade extracellular, hyaluronan-bound aggrecan aggregates in growth plate cartilage, and that lysosomal hydrolases are released from hypertrophic chondrocytes into growth plate cartilage via Ca2+-dependent lysosomal exocytosis. In this study we confirm that hypertrophic chondrocytes release hydrolases via lysosomal exocytosis in vitro and we show in vivo evidence for lysosomal exocytosis in hypertrophic chondrocytes during skeletal development. We show that lysosome-associated membrane protein 1 (LAMP1 is detected at the cell surface following in vitro treatment of epiphyseal chondrocytes with the calcium ionophore, ionomycin. Furthermore, we show that in addition to the lysosomal exocytosis markers, cathepsin D and β-hexosaminidase, ionomycin induces release of aggrecan- and hyaluronan-degrading activity from cultured epiphyseal chondrocytes. We identify VAMP-8 and VAMP7 as v-SNARE proteins with potential roles in lysosomal exocytosis in hypertrophic chondrocytes, based on their colocalisation with LAMP1 at the cell surface in secondary ossification centers in mouse tibiae. We propose that resorbing growth plate cartilage involves release of destructive hydrolases from hypertrophic chondrocytes, via lysosomal exocytosis.

  2. Antinutritional properties of plant lectins.

    Science.gov (United States)

    Vasconcelos, Ilka M; Oliveira, José Tadeu A

    2004-09-15

    Lectins are carbohydrate binding (glyco)proteins which are ubiquitous in nature. In plants, they are distributed in various families and hence ingested daily in appreciable amounts by both humans and animals. One of the most nutritionally important features of plant lectins is their ability to survive digestion by the gastrointestinal tract of consumers. This allows the lectins to bind to membrane glycosyl groups of the cells lining the digestive tract. As a result of this interaction a series of harmful local and systemic reactions are triggered placing this class of molecules as antinutritive and/or toxic substances. Locally, they can affect the turnover and loss of gut epithelial cells, damage the luminal membranes of the epithelium, interfere with nutrient digestion and absorption, stimulate shifts in the bacterial flora and modulate the immune state of the digestive tract. Systemically, they can disrupt lipid, carbohydrate and protein metabolism, promote enlargement and/or atrophy of key internal organs and tissues and alter the hormonal and immunological status. At high intakes, lectins can seriously threaten the growth and health of consuming animals. They are also detrimental to numerous insect pests of crop plants although less is presently known about their insecticidal mechanisms of action. This current review surveys the recent knowledge on the antinutritional/toxic effects of plant lectins on higher animals and insects. PMID:15302522

  3. A Novel Bifunctional Hybrid with Marine Bacterium Alkaline Phosphatase and Far Eastern Holothurian Mannan-Binding Lectin Activities

    Science.gov (United States)

    Balabanova, Larissa; Golotin, Vasily; Kovalchuk, Svetlana; Bulgakov, Alexander; Likhatskaya, Galina; Son, Oksana; Rasskazov, Valery

    2014-01-01

    A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25±5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens. PMID:25397876

  4. A novel bifunctional hybrid with marine bacterium alkaline phosphatase and Far Eastern holothurian mannan-binding lectin activities.

    Directory of Open Access Journals (Sweden)

    Larissa Balabanova

    Full Text Available A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25 ± 5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens.

  5. Role of the lectin complement pathway in kidney transplantation.

    Science.gov (United States)

    Farrar, Conrad A; Zhou, Wuding; Sacks, Steven H

    2016-10-01

    In the last 15 years two major advances in the role of complement in the kidney transplant have come about. The first is that ischaemia reperfusion injury and its profound effect on transplant outcome is dependent on the terminal product of complement activation, C5b-9. The second key observation relates to the function of the small biologically active fragments C3a and C5a released by complement activation in increasing antigen presentation and priming the T cell response that results in transplant rejection. In both cases local synthesis of C3 principally by the renal tubule cells plays an essential role that overshadows the role of the circulating pool of C3 generated largely by hepatocyte synthesis. More recent efforts have investigated the molecules expressed by renal tissue that can trigger complement activation. These have revealed a prominent effect of collectin-11 (CL-11), a soluble C-type lectin that is expressed in renal tissue and aligns with its major ligand L-fucose at sites of complement activation following ischaemic stress. Biochemical studies have shown that interaction between CL-11 and L-fucose results in complement activation by the lectin complement pathway, precisely targeting the innate immune response to the ischaemic tubule surface. Therapeutic approaches to reduce inflammatory and immune stimulation in ischaemic kidney have so far targeted C3 or its activation products and several are in clinical trials. The finding that lectin-fucose interaction is an important trigger of lectin pathway complement activation within the donor organ opens up further therapeutic targets where intervention could protect the donor kidney against complement. PMID:27286717

  6. Biological role of lectins: A review

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    K Kiran Kumar

    2012-01-01

    Full Text Available Lectins comprise a stracturally vary diverse class of proteins charecterized by their ability to selectively bind carbohydrate moieties of the glycoproteins of the cell surface. Lectins may be derived from plants, microbial or animal sources and may be soluble or membrane bound. Lectins is a tetramer made up of four nearly identical subunits. In human, lectins have been reported to cause food poisoning, hemolytic anemia, jaundice, digestive distress, protein and carbohydrate malabsorption and type I allergies. The present review focuses on the classification, structures, biological significance and application of lectins.

  7. Crystal structure of extracellular domain of human lectin-like transcript 1 (LLT1), the ligand for natural killer receptor-P1A.

    Science.gov (United States)

    Kita, Shunsuke; Matsubara, Haruki; Kasai, Yoshiyuki; Tamaoki, Takaharu; Okabe, Yuki; Fukuhara, Hideo; Kamishikiryo, Jun; Krayukhina, Elena; Uchiyama, Susumu; Ose, Toyoyuki; Kuroki, Kimiko; Maenaka, Katsumi

    2015-06-01

    Emerging evidence has revealed the pivotal roles of C-type lectin-like receptors (CTLRs) in the regulation of a wide range of immune responses. Human natural killer cell receptor-P1A (NKRP1A) is one of the CTLRs and recognizes another CTLR, lectin-like transcript 1 (LLT1) on target cells to control NK, NKT and Th17 cells. The structural basis for the NKRP1A-LLT1 interaction was limitedly understood. Here, we report the crystal structure of the ectodomain of LLT1. The plausible receptor-binding face of the C-type lectin-like domain is flat, and forms an extended β-sheet. The residues of this face are relatively conserved with another CTLR, keratinocyte-associated C-type lectin, which binds to the CTLR member, NKp65. A LLT1-NKRP1A complex model, prepared using the crystal structures of LLT1 and the keratinocyte-associated C-type lectin-NKp65 complex, reasonably satisfies the charge consistency and the conformational complementarity to explain a previous mutagenesis study. Furthermore, crystal packing and analytical ultracentrifugation revealed dimer formation, which supports a complex model. Our results provide structural insights for understanding the binding modes and signal transduction mechanisms, which are likely to be conserved in the CTLR family, and for further rational drug design towards regulating the LLT1 function. PMID:25826155

  8. Isolation, functional, and partial biochemical characterization of galatrox, an acidic lectin from Bothrops atrox snake venom

    Institute of Scientific and Technical Information of China (English)

    Elaine de Paula Mendonca-Franqueiro; Eliane Candiani Arantes; Marcelo Dias-Baruffi; Suely Vilela Sampaio; Raquel de Melo Alves-Paiva; Marco Aurélio Sartim; Daniel Roberto Callejon; Helder Henrique Paiva; Gilmara Ausech Antonucci; José César Rosa; Adélia Cristina Oliveira Cintra; Jo(a)o José Franco

    2011-01-01

    Snake venom lectins have been studied in regard to their chemical structure and biological functions. However, little is known about lectins isolated from Bothrops atrox snake venom. We report here the isolation and partial functional and biochemical characterization of an acidic glycanbinding protein called galatrox from this venom. This lectin was purified by affinity chromatography using a lactosyl-sepharose column, and its homogeneity and molecular mass were evaluated by high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. The purified galatrox was homogeneous and characterized as an acidic protein (pI 5.2) with a monomeric and dimeric molecular mass of 16.2 and 32.5 kDa, respectively. Alignment of N-terminal and internal amino acid sequences of galatrox indicated that this protein exhibits high homology to other C-type snake venom lectins. Galatrox showed optimal hemagglutinating activity at a concentration of 100 μg/ml and this effect was drastically inhibited by lactose, ethylenediaminetetraacetic acid, and heating, which confirmed galatrox's lectin activity. While galatrox failed to induce the same level of paw edema or mast cell degranulation as B. atrox crude venom, galatrox did alter cellular viability,which suggested that galatrox might contribute to venom toxicity by directly inducing cell death.

  9. A Novel Bifunctional Hybrid with Marine Bacterium Alkaline Phosphatase and Far Eastern Holothurian Mannan-Binding Lectin Activities

    OpenAIRE

    Larissa Balabanova; Vasily Golotin; Svetlana Kovalchuk; Alexander Bulgakov; Galina Likhatskaya; Oksana Son; Valery Rasskazov

    2014-01-01

    A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa...

  10. Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom

    Directory of Open Access Journals (Sweden)

    Fernandes Luiz

    2011-01-01

    Full Text Available Abstract Background Neutrophil migration to an inflamed site constitutes the first line of the innate immune response against invading microorganisms. Given the crucial role of endogenous lectins in neutrophil mobilization and activation, lectins from exogenous sources have often been considered as putative modulators of leukocyte function. Lectins purified from snake venom have been described as galactoside ligands that induce erythrocyte agglutination and platelet aggregation. This study evaluated human neutrophil migration and activation by C-type lectin BJcuL purified from Bothrops jararacussu venom. Results Utilizing fluorescence microscopy, we observed that biotinylated-BJcuL was evenly distributed on the neutrophil surface, selectively inhibited by D-galactose. Lectin was able to induce modification in the neutrophil morphology in a spherical shape for a polarized observed by optical microscopy and exposure to BJcuL in a Boyden chamber assay resulted in cell migration. After 30 minutes of incubation with BJcuL we found enhanced neutrophil functions, such as respiratory burst, zymozan phagocytosis and an increase in lissosomal volume. In addition, BJcuL delays late apoptosis neutrophils. Conclusion These results demonstrate that BJcuL can be implicated in a wide variety of immunological functions including first-line defense against pathogens, cell trafficking and induction of the innate immune response since lectin was capable of inducing potent neutrophil activation.

  11. Genetics Home Reference: mannose-binding lectin deficiency

    Science.gov (United States)

    ... Health Conditions mannose-binding lectin deficiency mannose-binding lectin deficiency Enable Javascript to view the expand/collapse ... PDF Open All Close All Description Mannose-binding lectin deficiency is a condition that affects the immune ...

  12. Lectin cDNA and transgenic plants derived therefrom

    Science.gov (United States)

    Raikhel, Natasha V.

    2000-10-03

    Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

  13. Gallus gallus aggrecan gene-based phylogenetic analysis of selected avian taxonomic groups.

    Science.gov (United States)

    Smith, Edward J; Shi, Li; Tu, Zhijian

    2005-05-01

    Mitochondrial DNA (mtDNA) sequences remain the most widely used for phylogenetic analysis in birds. A major limitation of mtDNA sequences, however, is that mitochondria genes are inherited as a single linkage group. Here we describe the use of a 540-bp DNA sequence corresponding to the G3 domain of Gallus gallus nuclear aggrecan gene (AGCI) for phylogenetic analysis of the main groups of Galliformes including Phasianidae, Numididae, and Odontophoridae. We also included species from Cracidae and Megapodiidae which are considered by some as Craciformes and others, including here as Galliformes. The uncorrected sequence divergence of the G3 fragments ranges from 1% among the grouses to 36% between some of the distant groups within Galliformes. These sequences contain 39-48% AT nucleotides and the ratios of transition versus transversion are above 1.5 in majority of the comparisons. Using G3 sequences from an Anseriform, Oxyura jamaicensis, as out-groups, phylogenetic trees were obtained using maximum parsimony and distance algorithms and bootstrap analyses. These trees were consistent with those described using Avian sarcoma and leucosis virus gag genes and those from amino acid sequences of hemoglobin and lysozyme c. Our data also support relationships among Galliformes which were defined using mtDNA sequences. In addition to the general support of the five main families of Galliformes, our data are also consistent with previous work that showed Francolinus africanus and Gallus gallus are in the same clade and that Tetraoninae is a well-supported monophyletic subfamily within Phasianidae. The results presented here suggest that the AGC1 sequences meet the criterion of novel nuclear DNA sequences that can be used to help resolve the relationships among Galliformes. PMID:16011000

  14. Molecular and biological characterization of a mannan-binding lectin from the holothurian Apostichopus japonicus.

    Science.gov (United States)

    Bulgakov, Aleksandr A; Eliseikina, Marina G; Petrova, Irina Yu; Nazarenko, Evgeny L; Kovalchuk, Svetlana N; Kozhemyako, Valery B; Rasskazov, Valery A

    2007-12-01

    To elucidate the origin and evolution of mannan-binding lectins (MBL), a new C-type lectin (CTL) specific for high-mannose glycans (MBL-AJ) was isolated from the coelomic plasma of the holothurian Apostichopus japonicus. MBL-AJ has oligomeric forms with identical 17-kDa subunits on SDS-PAGE. Among natural ligands, lectin hemagglutination activity was competitively inhibited by extracellular low-branched, but not high-branched, alpha-D-mannans isolated from marine halophilic bacteria and composed of alpha-1,2 and alpha-1,6 linked D-mannose residues. This suggests that the lectin interacts with backbone or inner side chain mannose residues, but not with terminal ones. The activity of the lectin was Ca(2+)-, pH-, and temperature-dependent. MBL-AJ cDNA was cloned from a holothurian coelomocyte cDNA library. The subunit of the mature protein has 159 amino acids and a single carbohydrate-recognition domain (CRD) of CTL. CRD contains a Glu-Pro-Asp amino acid sequence (EPN-motif) conserved for all known MBLs. A monospecific polyclonal antibody against MBL-AJ was obtained using the 34-kDa lectin dimer as an immunogen. The MBL-AJ has demonstrated immunochemical identity to the earlier isolated mannan-binding CTL from another holothurian, Cucumaria japonica. But a more interesting finding was cross-reactivity of MBL-AJ and human serum MBL detected by the antibody against MBL-AJ. Taking into consideration such MBL-AJ peculiarities as its carbohydrate specificity, the presence of a conserved region forming the mannose-binding site, common antigenic determinants with human MBL, and participation in defense reactions, it is possible that MBL-AJ belongs to the family of evolutionary conserved mannan-binding proteins. PMID:17890508

  15. Allergy-Protective Arabinogalactan Modulates Human Dendritic Cells via C-Type Lectins and Inhibition of NF-κB.

    Science.gov (United States)

    Peters, Marcus; Guidato, Patrick M; Peters, Karin; Megger, Dominik A; Sitek, Barbara; Classen, Birgit; Heise, Esther M; Bufe, Albrecht

    2016-02-15

    Arabinogalactan (AG) isolated from dust of a traditional farm prevents disease in murine models of allergy. However, it is unclear whether this polysaccharide has immune regulatory properties in humans. The aim of this study was to test the influence of AG on the immune-stimulating properties of human dendritic cells (DCs). Moreover, we sought to identify the receptor to which AG binds. AG was produced from plant callus tissue under sterile conditions to avoid the influence of pathogen-associated molecular patterns in subsequent experiments. The influence of AG on the human immune system was investigated by analyzing its impact on monocyte-derived DCs. To analyze whether the T cell stimulatory capacity of AG-stimulated DCs is altered, an MLR with naive Th cells was performed. We revealed that AG reduced T cell proliferation in a human MLR. In the search for a molecular mechanism, we found that AG binds to the immune modulatory receptors DC-specific ICAM-3 -: grabbing non integrin (DC-SIGN) and macrophage mannose receptor 1 (MMR-1). Stimulation of these receptors with AG simultaneously with TLR4 stimulation with LPS increased the expression of the E3 ubiquitin-protein ligase tripartite motif -: containing protein 21 and decreased the phosphorylation of NF-κB p65 in DCs. This led to a reduced activation profile with reduced costimulatory molecules and proinflammatory cytokine production. Blocking of MMR-1 or DC-SIGN with neutralizing Abs partially inhibits this effect. We conclude that AG dampens the activation of human DCs by LPS via binding to DC-SIGN and MMR-1, leading to attenuated TLR signaling. This results in a reduced T cell activation capacity of DCs. PMID:26746190

  16. A C-type lectin receptor pathway is responsible for the pathogenesis of acute cyclophosphamide-induced cystitis in mice.

    Science.gov (United States)

    Dejima, Takashi; Shibata, Kensuke; Yamada, Hisakata; Takeuchi, Ario; Hara, Hiromitsu; Eto, Masatoshi; Naito, Seiji; Yoshikai, Yasunobu

    2013-12-01

    Hemorrhagic cystitis often arises after cyclophosphamide (CYP) administration. As yet, however, the mechanism involved in its pathogenesis is unknown. In this study, it was found that the Fc receptor γ chain (FcRγ)- caspase recruitment domain-containing protein 9 (CARD9)-dependent pathway rather than the myeloid differentiation primary response gene 88 (MyD88)-dependent pathway is involved in the pathogenesis of acute CYP-induced cystitis in mice. Rapid and transient production of interleukin (IL)-6 and IL-1β was detected in the bladder at 4 hr, preceding IL-23 and IL-17A production and an influx of neutrophils, which reached a peak at 24 hr after injection. As assessed by weight, edema and neutrophil infiltration, cystitis was significantly attenuated in CARD9 knockout (KO) and FcRγKO mice, this attenuation being accompanied by impaired production of IL-1β, IL-6, IL-23 and IL-17A. The major source of IL-17A is the vesical γδ T cell population: IL-17AKO, CδKO and Tyk2KO mice showed little IL-17A production and reduced neutrophil infiltration in the bladder after CYP injection. These results suggest that FcRγ-CARD9-dependent production of proinflammatory cytokines such as IL-1β, IL-6, and IL-23 and the subsequent activation of IL-17A-producing γδ T cells are at least partly involved in the pathogenesis of acute CYP-induced cystitis in mice. PMID:24102807

  17. The Role of Syk/CARD9-Coupled C-Type Lectin Receptors in Immunity to Mycobacterium tuberculosis Infections

    Directory of Open Access Journals (Sweden)

    Mohlopheni Jackson Marakalala

    2010-01-01

    Full Text Available There is increasing interest in understanding the mechanisms underlying the interactions that occur between Mycobacterium tuberculosis and host innate immune cells. These cells express pattern recognition receptors (PRRs which recognise mycobacterial pathogen-associated molecular patterns (PAMPs and which can influence the host immune response to the infection. Although many of the PRRs appear to be redundant in the control of M. tuberculosis infection in vivo, recent discoveries have revealed a key, nonredundant, role of the Syk/CARD9 signalling pathway in antimycobacterial immunity. Here we review these discoveries, as well as recent data investigating the role of the Syk/CARD9-coupled PRRs that have been implicated in mycobacterial recognition, including Dectin-1 and Mincle.

  18. The Role of Syk/CARD9-Coupled C-Type Lectin Receptors in Immunity to Mycobacterium tuberculosis Infections

    OpenAIRE

    Mohlopheni Jackson Marakalala; Graham, Lisa M.; Brown, Gordon D.

    2010-01-01

    There is increasing interest in understanding the mechanisms underlying the interactions that occur between Mycobacterium tuberculosis and host innate immune cells. These cells express pattern recognition receptors (PRRs) which recognise mycobacterial pathogen-associated molecular patterns (PAMPs) and which can influence the host immune response to the infection. Although many of the PRRs appear to be redundant in the control of M. tuberculosis infection in vivo, recent discoveries have reve...

  19. Structural Changes in the Lectin Domain of CD23, the Low-Affinity IgE Receptor, upon Calcium Binding

    Energy Technology Data Exchange (ETDEWEB)

    Wurzburg, Beth A.; Tarchevskaya, Svetlana S.; Jardetzky, Theodore S. (NWU)

    2010-03-08

    CD23, the low-affinity receptor for IgE (Fc{var_epsilon}RII), regulates IgE synthesis and also mediates IgE-dependent antigen transport and processing. CD23 is a unique Fc receptor belonging to the C-type lectin-like domain superfamily and binds IgE in an unusual, non-lectin-like manner, requiring calcium but not carbohydrate. We have solved the high-resolution crystal structures of the human CD23 lectin domain in the presence and absence of Ca{sup 2+}. The crystal structures differ significantly from a previously determined NMR structure and show that calcium binding occurs at the principal binding site, but not at an auxiliary site that appears to be absent in human CD23. Conformational differences between the apo and Ca{sup 2+} bound structures suggest how IgE-Fc binding can be both calcium-dependent and carbohydrate-independent.

  20. Wide bandwidth nanomechanical assessment of murine cartilage reveals protection of aggrecan knock-in mice from joint-overuse.

    Science.gov (United States)

    Azadi, Mojtaba; Nia, Hadi Tavakoli; Gauci, Stephanie J; Ortiz, Christine; Fosang, Amanda J; Grodzinsky, Alan J

    2016-06-14

    Aggrecan loss in human and animal cartilage precedes clinical symptoms of osteoarthritis, suggesting that aggrecan loss is an initiating step in cartilage pathology. Characterizing early stages of cartilage degeneration caused by aging and overuse is important in the search for therapeutics. In this study, atomic force microscopy (AFM)-based force-displacement micromechanics, AFM-based wide bandwidth nanomechanics (nanodynamic), and histologic assessments were used to study changes in distal femur cartilage of wildtype mice and mice in which the aggrecan interglobular domain was mutated to make the cartilage aggrecanase-resistant. Half the animals were subjected to voluntary running-wheel exercise of varying durations. Wildtype mice at three selected age groups were compared. While histological assessment was not sensitive enough to capture any statistically significant changes in these relatively young populations of mice, micromechanical assessment captured changes in the quasi-equilibrium structural-elastic behavior of the cartilage matrix. Additionally, nanodynamic assessment captured changes in the fluid-solid poroelastic behavior and the high frequency stiffness of the tissue, which proved to be the most sensitive assessment of changes in cartilage associated with aging and joint-overuse. In wildtype mice, aging caused softening of the cartilage tissue at the microscale and at the nanoscale. Softening with increased animal age was found at high loading rates (frequencies), suggesting an increase in hydraulic permeability, with implications for loss of function pertinent to running and impact-injury. Running caused substantial changes in fluid-solid interactions in aggrecanase-resistant mice, suggestive of tissue degradation. However, higher nanodynamic stiffness magnitude and lower hydraulic permeability was observed in running aggrecanase-resistant mice compared to running wildtype controls at the same age, thereby suggesting protection from joint

  1. [Fractionation of lymphocytes using affinity chromatography with 9 lectins].

    Science.gov (United States)

    de Dios, I; Manso, M; López-Borrasca, A

    1983-12-01

    Lymphocyte subclasses from normal peripheral blood have been fractionated by affinity chromatography with lectins. Concanavalin A (Con A), Lens culinaris lectin (LC), Pisum sativum lectin (PS), Phaseolus vulgaris lectin (PHA), Dolichos biflours lectin (DB), Glicine max lectin (SBA), Ricinus communis lectin (RCA II), Tetragonolobus purpureus lectin (TP) and Triticum vulgaris lectin (WGA), were coupled to Sepharose 6MB, and lymphocytes labelled with 125I were eluted through the chromatographic columns. The binding of lymphocytes to WGA and SBA lectins was 32% and 13% respectively. The binding to the other lectins tested were found to be between 32% and 13%. When solutions of increasing concentrations of specific sugar were added to the columns a progressive elution of bound lymphocytes was observed. These results indicate the existence of a large range of lymphocyte subclasses, with different binding capacity to lectins, which was a function of the receptor number or/and receptor affinity to each lectin. Furthermore, these two parameters were found to vary in each functional population. Even though all the lymphocytes had lectin receptors, T lymphocytes showed higher affinity for Con A, PHA and TP lectins, while B lymphocytes appeared to be more specific for LC, PS, SBA, DB, RCAII and WGA lectins. PMID:6675094

  2. Characterization and Localization of Citrullinated Proteoglycan Aggrecan in Human Articular Cartilage.

    Directory of Open Access Journals (Sweden)

    Tibor T Glant

    Full Text Available Rheumatoid arthritis (RA is an autoimmune disease of the synovial joints. The autoimmune character of RA is underscored by prominent production of autoantibodies such as those against IgG (rheumatoid factor, and a broad array of joint tissue-specific and other endogenous citrullinated proteins. Anti-citrullinated protein antibodies (ACPA can be detected in the sera and synovial fluids of RA patients and ACPA seropositivity is one of the diagnostic criteria of RA. Studies have demonstrated that RA T cells respond to citrullinated peptides (epitopes of proteoglycan (PG aggrecan, which is one of the most abundant macromolecules of articular cartilage. However, it is not known if the PG molecule is citrullinated in vivo in human cartilage, and if so, whether citrulline-containing neoepitopes of PG (CitPG can contribute to autoimmunity in RA.CitPG was detected in human cartilage extracts using ACPA+ RA sera in dot blot and Western blot. Citrullination status of in vitro citrullinated recombinant G1 domain of human PG (rhG1 was confirmed by antibody-based and chemical methods, and potential sites of citrullination in rhG1 were explored by molecular modeling. CitPG-specific serum autoantibodies were quantified by enzyme-linked immunosorbent assays, and CitPG was localized in osteoarthritic (OA and RA cartilage using immunohistochemistry.Sera from ACPA+ RA patients reacted with PG purified from normal human cartilage specimens. PG fragments (mainly those containing the G1 domain from OA or RA cartilage extracts were recognized by ACPA+ sera but not by serum from ACPA- individuals. ACPA+ sera also reacted with in vitro citrullinated rhG1 and G3 domain-containing fragment(s of PG. Molecular modeling suggested multiple sites of potential citrullination within the G1 domain. The immunohistochemical localization of CitPG was different in OA and RA cartilage.CitPG is a new member of citrullinated proteins identified in human joints. CitPG could be found in

  3. Recent Progress in Lectin-Based Biosensors

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    Baozhen Wang

    2015-12-01

    Full Text Available This article reviews recent progress in the development of lectin-based biosensors used for the determination of glucose, pathogenic bacteria and toxins, cancer cells, and lectins. Lectin proteins have been widely used for the construction of optical and electrochemical biosensors by exploiting the specific binding affinity to carbohydrates. Among lectin proteins, concanavalin A (Con A is most frequently used for this purpose as glucose- and mannose-selective lectin. Con A is useful for immobilizing enzymes including glucose oxidase (GOx and horseradish peroxidase (HRP on the surface of a solid support to construct glucose and hydrogen peroxide sensors, because these enzymes are covered with intrinsic hydrocarbon chains. Con A-modified electrodes can be used as biosensors sensitive to glucose, cancer cells, and pathogenic bacteria covered with hydrocarbon chains. The target substrates are selectively adsorbed to the surface of Con A-modified electrodes through strong affinity of Con A to hydrocarbon chains. A recent topic in the development of lectin-based biosensors is a successful use of nanomaterials, such as metal nanoparticles and carbon nanotubes, for amplifying output signals of the sensors. In addition, lectin-based biosensors are useful for studying glycan expression on living cells.

  4. Agglutination of Helicobacter pylori coccoids by lectins

    Institute of Scientific and Technical Information of China (English)

    Mar Mar Khin; Jie Song Hua; Hah Cong Ng; Bow Ho; Torkel Wadstrorr

    2000-01-01

    AIM To study the agglutination pattern of Helicobacter pylori coccoid and spiral forms.METHODS Assays of agglutination and agglutination inhibition were applied using fifteen commercial lectins. RESULTS Strong agglutination was observed with mannose-specific Concanavalin A (Con A ),fucose-specific Tetragonolobus purpureas ( Lotus A ) and N-acetyl glucosamine-specific Triticum vulgaris (WGA) lectins. Mannose and fucose specific lectins were reactive with all strains of H. pylori coccoids as compared to the spirals. Specific carbohydrates, glycoproteins and mucin were shown to inhibit H. pylori lectin-agglutination reactions. Pre-treatment of the bacterial cells with formalin and sulphuric acid did not alter the agglutination patterns with lectins. However, sodium periodate treatment of bacterial cells were shown to inhibit agglutination reaction with Con A, Lotus A and WGA lectins. On the contrary, enzymatic treatment of coccoids and spirals did not show marked inhibition of H. pylori-lectin agglutination. Interestingly, heating of H.pylori cells at 60℃ for 1 hour was shown to augment the agglutination with all of the lectins tested. CONCLUSION The considerable differences in lectin agglutination patterns seen among the two differentiated forms of H. pylori might be attributable to the structural changes during theevents of morphological transformation,resulting in exposing or masking some of the sugar residues on the cell surface. Possibility of various sugar residues on the cell wall of the coccoids may allow them to bind to different carbohydrate receptors on gastric mucus and epithelial cells. The coccoids with adherence characteristics like the spirals could aid in the pathogenic process of Helicobacter infection.This may probably lead to different clinical outcome of H. pylori associated gastroduodenal disease.

  5. Assessing biosafety of GM plants containing lectins

    DEFF Research Database (Denmark)

    Poulsen, Morten; Pedersen, Jan W.

    2010-01-01

    The introduction of genetic engineering has already shown its benefits in transferring genes into crop plants and conferring resistance towards pests. Most of these crop plants on the market have been transformed with the cry genes from Bacillus species, conferring resistance towards certain...... of the lectins that are potentially useful for insect resistance will pose no health risk in genetically modified (GM) plants. Since some lectins are known for their toxicity to humans, the insertion of lectin genes in food crop plants will have to be assessed carefully. It is expected that in some cases...

  6. Lectin binding in normal donkey eyeball

    Directory of Open Access Journals (Sweden)

    Khaled Aly

    2013-10-01

    Full Text Available In the present study, the distribution of various sugar residues in the eyeball tissues of sexually mature donkey was examined by employing fluorescein isothiocyanate-conjugated lectins. Our results revealed the presence of mannose (labeled by lectins ConA, galactose (labeled by PNA, GSAI, ECA, GalNAc (labeled by SBA, VVA, and GlcNAc (labeled by WGA residues in the donkey ocular tissues. The epithelium and stroma of the ocular tissues were labeled with mannose (ConA and GlcNAc (WGA binding lectins. Binding sites for WGA and PNA to the rod and cone cells of the retina were evident. The lectins Con A, WGA and GSAI are bound strongly to the endothelium of blood vessels and to smooth muscle cells of the iris. In conclusion, the findings of the present study clearly indicate that the donkey eyeball contains a wide range of glycoconjugates (bearing mannosyl, galactosyl and glucosly residues, and it lacks fucosyl residues.

  7. Lectin histochemistry of pregnant rat uterine tissues.

    OpenAIRE

    Peel, S; Bulmer, J. N.

    1996-01-01

    Glycoconjugate residues were examined in the rat uterus at days 10, 12 and 15 of pregnancy using 17 biotinylated lectins with specificities for a variety of carbohydrate moieties. A wide variety of glycoconjugate residues were detected in the cytoplasm of some antimesometrial and mesometrial decidual cells with lectins from Canavalia ensiformis (Con A), Lens culinaris (LcH) culinaris (LcH), Pisum sativum (PSA), Griffonia simplicifolia II (GS-II), Triticum vulgaris (WGA), Griffonia simplicifol...

  8. The Lectin Pathway of Complement and Biocompatibility

    DEFF Research Database (Denmark)

    Hein, Estrid; Garred, Peter

    2015-01-01

    activation, the coagulation system and the complement system. The complement system is an important part of the initial immune response and consists of fluid phase molecules in the blood stream. Three different activation pathways can initiate the complement system, the lectin, the classical and the...... been broadly documented. However, the specific role of lectin pathway and the pattern recognition molecules initiating the pathway has only been transiently investigated. Here we review the current data on the field....

  9. Isolation and characteristics of autoreactive T cells spe cific to aggrecan G1 domain from rheumatoid arthritis patients

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Our previous work showed that the cartilage proteo glycan aggrecan could induce an erosive polyarthritis and spondylitis in BALB/c mice and the G1 globular domain of the aggrecan (G1) contained the arthritogenic region. To elucidate whether autoreactive T cells to G1 are ex pressed in rheumatoid arthritis patients, we analyzed the frequency of human G1-specific T cells in the peripheral blood of five rheumatoid arthritis patients and tried to establish G1-reactive T cell lines from these rheumatoid arthritis patients. The results showed that the Gl-specific T cells in PBL were detectable at the range of 4.97 + 0.5 × 10-6 in peripheral blood lymphocytes. We have also generated 15 G1-specific T lymphocyte lines from these pateints with a standard split-well method. All these cells expressed fine specificity to human recombinant G1, but not to unrelated antigen. All the 15 lines expressed a pan T cell marker and 13 of them selectively used the αβ T cell receptor. Two of them used γδ T cell receptor. The 13 of these T cell lines was CD4 positive. One line expressed CD8. One line expressed both CD4 and CD8. More over, 14 out of 15 lines expressed the Th-1 cytokine profile, characterized by interferon-γ positivity and IL-4 negativ ity. No Th-2 type cell line was generated. These data provide strong evidence in favor of the presence of autore active T cells in the rheumatoid arthritis pateints. What is the mechanism(s) that these autoreactive T cells attack self-target and whether these Gl-specific, Th-1 type T cell lines can induce arthritis in immune deficiency mice are currently under investigation.

  10. Sugared biomaterial binding lectins: achievements and perspectives.

    Science.gov (United States)

    Bojarová, P; Křen, V

    2016-07-19

    Lectins, a distinct group of glycan-binding proteins, play a prominent role in the immune system ranging from pathogen recognition and tuning of inflammation to cell adhesion or cellular signalling. The possibilities of their detailed study expanded along with the rapid development of biomaterials in the last decade. The immense knowledge of all aspects of glycan-lectin interactions both in vitro and in vivo may be efficiently used in bioimaging, targeted drug delivery, diagnostic and analytic biological methods. Practically applicable examples comprise photoluminescence and optical biosensors, ingenious three-dimensional carbohydrate microarrays for high-throughput screening, matrices for magnetic resonance imaging, targeted hyperthermal treatment of cancer tissues, selective inhibitors of bacterial toxins and pathogen-recognising lectin receptors, and many others. This review aims to present an up-to-date systematic overview of glycan-decorated biomaterials promising for interactions with lectins, especially those applicable in biology, biotechnology or medicine. The lectins of interest include galectin-1, -3 and -7 participating in tumour progression, bacterial lectins from Pseudomonas aeruginosa (PA-IL), E. coli (Fim-H) and Clostridium botulinum (HA33) or DC-SIGN, receptors of macrophages and dendritic cells. The spectrum of lectin-binding biomaterials covered herein ranges from glycosylated organic structures, calixarene and fullerene cores over glycopeptides and glycoproteins, functionalised carbohydrate scaffolds of cyclodextrin or chitin to self-assembling glycopolymer clusters, gels, micelles and liposomes. Glyconanoparticles, glycan arrays, and other biomaterials with a solid core are described in detail, including inorganic matrices like hydroxyapatite or stainless steel for bioimplants. PMID:27075026

  11. The biological role of hemolymph lectins in Episesarma tetragonum

    Directory of Open Access Journals (Sweden)

    VR Devi

    2013-12-01

    Full Text Available The hemolymph of the mangrove crab, E. tetragonum contains lectins specific for NeuGcα 2, 3 Gal β1-4 GluNAc β1 linkage and O-acetyl sialic acids. The role of sialic acid specific lectins on natural immunity of the crab is studied by using several kinds of mammalian erythrocytes as pathogen model. Injection of erythrocytes showing differential agglutinability with the lectins, induced augmentation of hemagglutinating activity suggesting an increase in the circulating lectins. A significant correlation was observed between in vivo clearances of exogenous erythrocytes with the extent of erythrocyte agglutination by the lectins. Another correlation was observed between the susceptibility of erythrocytes to lectin dependent hemocyte mediated hemolysis and the extent of lectin mediated erythrocyte agglutination. This study documents that opsonization of foreign pathogen by the native lectins is an important step in hemocyte recognition, hemolysis and clearance of the pathogen.

  12. Porifera Lectins: diversity, physiological roles and biotechnological potential

    OpenAIRE

    Johan Gardères; Marie-Lise Bourguet-Kondracki; Bojan Hamer; Renato Batel; Schröder, Heinz C.; Müller, Werner E. G.

    2015-01-01

    An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial system...

  13. Lectin interactions with alpha-galactosylated xenoantigens

    DEFF Research Database (Denmark)

    Kirkeby, Svend; Moe, Dennis

    2002-01-01

    alpha-Galactosylated xenoantigens (Galalpha1-3Galbeta1-4GlcNAcbeta1 and Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc) are often detected with the alpha-Gal specific lectin Griffonia simplicifolia 1 isolectin B4 (GS1 B4). However, this lectin exhibits a broad and variable specificity for......-galactophilic lectins to alpha-galactosylated neoglycoproteins. The lectins were: Euonymus europaeus agglutinin (EEA), Griffonia simplicifolia 1 isolectin B4 (GS1 B4), Maclura pomifera agglutinin (MPA) and Pseudomonas aeruginosa agglutinin (PA-IL). Although both GS1 B4 and MPA strongly bound glycoconjugates terminating...... in Gal there seems to be some differentiation in their sugar binding preferences. MPA was the only lectin that showed high affinity for the pentasaccharide Galalpha1-3Galbeta1-4GlcNAcbeta1-3Galbeta1-4Glc and for the Galalpha-glycans on non-primate thyroglobulin. The length of the xenoantigenic...

  14. 21 CFR 864.9550 - Lectins and protectins.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Lectins and protectins. 864.9550 Section 864.9550 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... and Blood Products § 864.9550 Lectins and protectins. (a) Identification. Lectins and protectins...

  15. Bacteriostatic activity of con a lectin from Canavalia ensiformis

    Directory of Open Access Journals (Sweden)

    S.R. Kulkarni *

    2013-12-01

    Full Text Available The aim of this work was to explore the therapeutic applications of Con A lectin from Canavalia ensiformis and to explore its antibacterial activity. Activity of lectin was quantified by their ability to agglutinate erythrocytes using Hemagglutination assay. Characterization and purity of Con A lectin was evaluated by using SDS-PAGE analysis. The reversal of hemagglutination activity of lectin was evaluated by using the sugars namely; mannose, galactose, lactose, fructose, glucose. The antibacterial activity of lectins was tested against Streptococcus mutans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli using pour plate method. Amoxycillin was used as standard. At 250mg/ml concentration Con A lectin showed good bacteriostatic activity.The aim of this work was to explore the therapeutic applications of Con A lectin from Canavalia ensiformis and to explore its antibacterial activity. Activity of lectin was quantified by their ability to agglutinate erythrocytes using Hemagglutination assay. Characterization and purity of Con A lectin was evaluated by using SDS-PAGE analysis. The reversal of hemagglutination activity of lectin was evaluated by using the sugars namely; mannose, galactose, lactose, fructose, glucose. The antibacterial activity of lectins was tested against Streptococcus mutans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli using pour plate method. Amoxycillin was used as standard. At 250mg/ml concentration Con A lectin showed good bacteriostatic activity.

  16. Effect of lectins on mouse peritoneal macrophage phagocytic activity.

    Science.gov (United States)

    Maldonado, G; Porras, F; Fernández, L; Vázquez, L; Zenteno, E

    1994-11-01

    We studied the in vitro ability of lectin-treated murine peritoneal macrophages to attach and phagocytize particulate antigens. Glucose and mannose specific lectins such as Con-A and lentil lectin, as well as complex lactosamine residues specific lectins, such as Phaseolus vulgaris var. cacahuate and Phaseolus coccineus var. alubia, increased the macrophage phagocytic activity towards heterologous erythrocytes, whereas peanut agglutinin, a galactose-specific lectin, diminished the macrophage phagocytic activity. These results suggest that a galactose-N-acetyl-D galactosamine-containing structure could participate as negative modulator of the phagocytic activity. PMID:7851961

  17. Age-related changes in the sulphation of the chondroitin sulphate linkage region from human articular cartilage aggrecan.

    Science.gov (United States)

    Lauder, R M; Huckerby, T N; Brown, G M; Bayliss, M T; Nieduszynski, I A

    2001-09-01

    The chondroitin sulphate (CS) linkage regions have been isolated from human articular cartilage aggrecan (from 10- to 72-year-olds) by chondroitin ABC endolyase digestion and size-exclusion chromatography. Linkage region hexasaccharides have been characterized and their abundance estimated by high-pH anion-exchange chromatography. The basic structure for the CS linkage region oligosaccharides identified from human aggrecan is as follows: DeltaUA(beta1-3)GalNAc[0S/4S/6S](beta1-4)GlcA(beta1-3)Gal[0S/6S](beta1-3)Gal(beta1-4)Xyl, where DeltaUA represents 4,5-unsaturated hexuronic acid, 4S and 6S represent an O-ester sulphate group on C-4 and C-6 respectively, and 0S represents zero sulphation. There are significant age-related changes in the abundance of the various N-acetylgalactosamine (GalNAc) sulphation forms identified, occurring up to approx. 20 years old. During the period from 10 to 20 years old the level of GalNAc 6-sulphation at the linkage region increases from approx. 43% to approx. 75%, while there is a corresponding reduction in unsulphated (approx. 30% to approx. 20%) and 4-sulphated (approx. 25% to approx. 6%) GalNAc residues. There is also an increase in the incidence of linkage region galactose 6-sulphation (approx. 2% to approx. 10%) which was only observed in linkage regions with GalNAc 6-sulphation. Beyond 20 years old there are few changes in the relative abundance of these GalNAc sulphation variants; however, there is a slight increase in the abundance of 6-sulphation between approx. 20 years old and approx. 40 years old and a slight decrease in its abundance beyond approx. 40 years old. Our data show that in the majority of chains from tissues of all ages the GalNAc residue closest to the linkage region is 6-sulphated, but the level of GalNAc 6-sulphation within the linkage region is lower than the average level observed within the repeat region. PMID:11513754

  18. ADAMTS4 and ADAMTS5 Knockout Mice Are Protected from Versican but Not Aggrecan or Brevican Proteolysis during Spinal Cord Injury

    OpenAIRE

    Kadir Demircan; Vehap Topcu; Tomoyuki Takigawa; Sumeyya Akyol; Tomoko Yonezawa; Gulfer Ozturk; Veli Ugurcu; Rukiye Hasgul; M. Ramazan Yigitoglu; Omer Akyol; McCulloch, Daniel R.; Satoshi Hirohata

    2014-01-01

    The chondroitin sulfate proteoglycans (CSPGs) aggrecan, versican, and brevican are large aggregating extracellular matrix molecules that inhibit axonal growth of the mature central nervous system (CNS). ADAMTS proteoglycanases, including ADAMTS4 and ADAMTS5, degrade CSPGs, representing potential targets for ameliorating axonal growth-inhibition by CSPG accumulation after CNS injury. We investigated the proteolysis of CSPGs in mice homozygous for Adamts4 or Adamts5 null alleles after spinal co...

  19. Lectins from tropical sponges. Purification and characterization of lectins from genus Aplysina.

    Science.gov (United States)

    Miarons, P B; Fresno, M

    2000-09-22

    Only a few animal phyla have been screened for the presence and distribution of lectins. Probably the most intensively studied group is the mollusk. In this investigation, 22 species from 12 families of tropical sponges collected in Los Roques National Park (Venezuela) were screened for the presence of lectins. Nine saline extracts exhibited strong hemagglutinating activity against pronase-treated hamster red blood cells; five of these reacted against rabbit red blood cells, four with trypsin-treated bovine red blood cells, and five with human red blood cells regardless of the blood group type. Extracts from the three species studied from genus Aplysina (archeri, lawnosa, and cauliformis) were highly reactive and panagglutinating against the panel of red blood cells tested. The lectins from A. archeri and A. lawnosa were purified to homogeneity by ammonium sulfate fractionation, affinity chromatography on p-aminobenzyl-beta-1-thiogalactopyranoside-agarose, and gel filtration chromatography. Both lectins exhibited a native molecular mass of 63 kDa and by SDS-polyacrylamide gel electrophoresis under reducing conditions have an apparent molecular mass of 16 kDa, thus suggesting they occur as homotetramers. The purified lectins contain 3-4 mol of divalent cation per molecule, which are essential for their biological activity. Hapten inhibition of hemagglutination was carried out to define the sugar binding specificity of the purified A. archeri lectin. The results indicate a preference of the lectin for nonreducing beta-linked d-Gal residues being the best inhibitors of red blood cells binding methyl-beta-d-Gal and thiodigalactoside (Gal beta 1-4-thiogalactopyranoside). The behavior of several glycans on immobilized lectin affinity chromatography confirmed and extended the specificity data obtained by hapten inhibition. PMID:10852905

  20. IDENTIFICATION OF LECTINS OF ZEA MAYS RAW MATERIAL AND THE STUDY OF LECTIN ACTIVITY

    Directory of Open Access Journals (Sweden)

    Karpiuk UV

    2013-03-01

    Full Text Available The aime of the study was to identify lectins in the Zea mays raw material: roots, stems, heads, leaves and corn silk and study their activity. Lectins activity has been studied using the biological method of ratuserytroagglutination. This method is based on formation of aggregates of lectins and rats erythrocytes. The activity unit was the floor amount of lectins that agglutinate erythrocytes. The protein nature of extracts that agglutinate has been determined using Bradford method. The lectins activity of Zea mays roots was 6,21±0,11 unit/mg of protein; of heads – 2,61±0,17 unit/mg of protein; of leaves – 0,62 ±0,05 unit/mg of protein; of corn silk – 1,06±0,08 unit/mg of protein; of stems – 0,97±0,09 unit/mg of protein. The greatest lectins activity was in leaves, stems and corn silk.

  1. ADAMTS4 and ADAMTS5 Knockout Mice Are Protected from Versican but Not Aggrecan or Brevican Proteolysis during Spinal Cord Injury

    Directory of Open Access Journals (Sweden)

    Kadir Demircan

    2014-01-01

    Full Text Available The chondroitin sulfate proteoglycans (CSPGs aggrecan, versican, and brevican are large aggregating extracellular matrix molecules that inhibit axonal growth of the mature central nervous system (CNS. ADAMTS proteoglycanases, including ADAMTS4 and ADAMTS5, degrade CSPGs, representing potential targets for ameliorating axonal growth-inhibition by CSPG accumulation after CNS injury. We investigated the proteolysis of CSPGs in mice homozygous for Adamts4 or Adamts5 null alleles after spinal cord injury (SCI. ADAMTS-derived 50–60 kDa aggrecan and 50 kDa brevican fragments were observed in Adamts4−/−, Adamts5−/−, and wt mice but not in the sham-operated group. By contrast Adamts4−/− and Adamts5−/− mice were both protected from versican proteolysis with an ADAMTS-generated 70 kDa versican fragment predominately observed in WT mice. ADAMTS1, ADAMTS9, and ADAMTS15 were detected by Western blot in Adamts4−/− mice’ spinal cords after SCI. Immunohistochemistry showed astrocyte accumulation at the injury site. These data indicate that aggrecan and brevican proteolysis is compensated in Adamts4−/− or Adamts5−/− mice by ADAMTS proteoglycanase family members but a threshold of versican proteolysis is sensitive to the loss of a single ADAMTS proteoglycanase during SCI. We show robust ADAMTS activity after SCI and exemplify the requirement for collective proteolysis for effective CSPG clearance during SCI.

  2. Lectin modified macroporous cryogels for glycoprotein analysis

    Czech Academy of Sciences Publication Activity Database

    Křenková, Jana; Václavek, Tomáš; Foret, František

    Salzburg: Society of Analytical Chemistry, 2014. P203-FF-TU. [International Symposium on Chromatography /30./. 14.09.2014-18.09.2014, Salzburg] R&D Projects: GA ČR(CZ) GA14-06319S Grant ostatní: GA AV ČR(CZ) M200311201 Institutional support: RVO:68081715 Keywords : monolith * cryogel * lectin Subject RIV: CB - Analytical Chemistry, Separation

  3. Possible role of lectins in mycoparasitism.

    OpenAIRE

    Elad, Y; Barak, R; Chet, I

    1983-01-01

    Lectin activity in a host-mycoparasite relationship was demonstrated with Rhizoctonia solani and Trichoderma harzianum. Attachment of O but not A and B erythrocytes to hyphae occurred on R. solani but not on its mycoparasite. This phenomenon, which was Ca2+ and Mn2+ dependent, was prevented by galactose, present in T. harzianum cell walls, and by fucose.

  4. The search for lectin isolated from the mycelial cultures of Laetiporus sulphureus

    Directory of Open Access Journals (Sweden)

    Grażyna Końska

    2014-08-01

    Full Text Available This study proved the presence of lectin in mycelial cultures of Laetiporus sulphureus. Lectin was excreted into the medium and its erythroagglutinating activity was not high. No active lectin was detected in hyphae using both extraction and immunofluorescence method. Comparative studies based on immunological methods indicate~ that the lectin synthesised in vitro differed from the lectin produced in fruit-bodies.

  5. The search for lectin isolated from the mycelial cultures of Laetiporus sulphureus

    OpenAIRE

    Grażyna Końska; Jean Guillot; Stanisław Kohlmünzer; Mireille Damez; Kinga Rakoczy

    2014-01-01

    This study proved the presence of lectin in mycelial cultures of Laetiporus sulphureus. Lectin was excreted into the medium and its erythroagglutinating activity was not high. No active lectin was detected in hyphae using both extraction and immunofluorescence method. Comparative studies based on immunological methods indicate~ that the lectin synthesised in vitro differed from the lectin produced in fruit-bodies.

  6. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    Directory of Open Access Journals (Sweden)

    Sondergaard Bodil-Cecilie

    2009-12-01

    Full Text Available Abstract Background Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP- and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. Findings Bovine cartilage explants were cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM and tumor necrosis factor alpha (TNFα. In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo-epitope specific immunoassays; (1 sandwich 342FFGVG-G2 ELISA, (2 competition NITEGE373ELISA (3 sandwich G1-NITEGE373 ELISA (4 competition 374ARGSV ELISA, and (5 sandwich 374ARGSV-G2 ELISA all detecting aggrecan fragments, and (6 sandwich CTX-II ELISA, detecting C-telopeptides of type II collagen. We found that (1 aggrecanase-derived aggrecan fragments are released in the early (day 2-7 and mid phase (day 9-14 into the supernatant from bovine explants cultures stimulated with catabolic cytokines, (2 the release of NITEGE373 neo-epitopes are delayed compared to the corresponding 374ARGSV fragments, (3 the MMP inhibitor GM6001 did not reduce the release of aggrecanase-derived fragment, but induced a further delay in the release of these fragments, and finally (4 the MMP-derived aggrecan and type II collagen fragments were released in the late phase (day 16-21 only. Conclusion Our data support the model, that aggrecanases and MMPs act independently in the processing of the aggrecan molecules, and furthermore that suppression of MMP-activity had little if

  7. A profile of protein-protein interaction: Crystal structure of a lectin-lectin complex.

    Science.gov (United States)

    Surya, Sukumaran; Abhilash, Joseph; Geethanandan, Krishnan; Sadasivan, Chittalakkottu; Haridas, Madhathilkovilakathu

    2016-06-01

    Proteins may utilize complex networks of interactions to create/proceed signaling pathways of highly adaptive responses such as programmed cell death. Direct binary interactions study of proteins may help propose models for protein-protein interaction. Towards this goal we applied a combination of thermodynamic kinetics and crystal structure analyses to elucidate the complexity and diversity in such interactions. By determining the heat change on the association of two galactose-specific legume lectins from Butea monosperma (BML) and Spatholobus parviflorus (SPL) belonging to Fabaceae family helped to compute the binding equilibrium. It was extended further by X-ray structural analysis of BML-SPL binary complex. In order to chart the proteins interacting mainly through their interfaces, identification of the nature of forces which stabilized the association of the lectin-lectin complex was examined. Comprehensive analysis of the BMLSPL complex by isothermal titration calorimetry and X-ray crystal structure threw new light on the lectin-lectin interactions suggesting of their use in diverse areas of glycobiology. PMID:26945504

  8. Lessons learned from mice deficient in lectin complement pathway molecules

    DEFF Research Database (Denmark)

    Genster, Ninette; Takahashi, Minoru; Sekine, Hideharu;

    2014-01-01

    differences in the genetic arrangements of murine and human orthologues of lectin pathway molecules, the knockout mice have proven to be valuable models to explore the effect of deficiency states in humans. In addition, new insight and unexpected findings on the diverse roles of lectin pathway molecules in......The lectin pathway of the complement system is initiated when the pattern-recognition molecules, mannose-binding lectin (MBL), ficolins or collectin-11, bind to invading pathogens or damaged host cells. This leads to activation of MBL/ficolin/collectin-11 associated serine proteases (MASPs), which...... in turn activate downstream complement components, ultimately leading to elimination of the pathogen. Mice deficient in the key molecules of lectin pathway of complement have been generated in order to build knowledge of the molecular mechanisms of the lectin pathway in health and disease. Despite...

  9. Algal lectins as promising biomolecules for biomedical research.

    Science.gov (United States)

    Singh, Ram Sarup; Thakur, Shivani Rani; Bansal, Parveen

    2015-02-01

    Lectins are natural bioactive ubiquitous proteins or glycoproteins of non-immune response that bind reversibly to glycans of glycoproteins, glycolipids and polysaccharides possessing at least one non-catalytic domain causing agglutination. Some of them consist of several carbohydrate-binding domains which endow them with the properties of cell agglutination or precipitation of glycoconjugates. Lectins are rampant in nature from plants, animals and microorganisms. Among microorganisms, algae are the potent source of lectins with unique properties specifically from red algae. The demand of peculiar and neoteric biologically active substances has intensified the developments on isolation and biomedical applications of new algal lectins. Comprehensively, algal lectins are used in biomedical research for antiviral, antinociceptive, anti-inflammatory, anti-tumor activities, etc. and in pharmaceutics for the fabrication of cost-effective protein expression systems and nutraceutics. In this review, an attempt has been made to collate the information on various biomedical applications of algal lectins. PMID:23855360

  10. Specificity analysis of lectins and antibodies using remodeled glycoproteins

    OpenAIRE

    Iskratsch, Thomas; Braun, Andreas; Paschinger, Katharina; Wilson, Iain B. H.

    2009-01-01

    Due to their ability to bind specifically to certain carbohydrate sequences, lectins are a frequently used tool in cytology, histology, and glycan analysis but also offer new options for drug targeting and drug delivery systems. For these and other potential applications, it is necessary to be certain as to the carbohydrate structures interacting with the lectin. Therefore, we used glycoproteins remodeled with glycosyltransferases and glycosidases for testing specificities of lectins from Ale...

  11. Insights into Animal and Plant Lectins with Antimicrobial Activities

    OpenAIRE

    Renata de Oliveira Dias; Leandro dos Santos Machado; Ludovico Migliolo; Octavio Luiz Franco

    2015-01-01

    Lectins are multivalent proteins with the ability to recognize and bind diverse carbohydrate structures. The glyco -binding and diverse molecular structures observed in these protein classes make them a large and heterogeneous group with a wide range of biological activities in microorganisms, animals and plants. Lectins from plants and animals are commonly used in direct defense against pathogens and in immune regulation. This review focuses on sources of animal and plant lectins, describing...

  12. ON VASCULAR STENOSIS, RESTENOSIS AND MANNOSE BINDING LECTIN.

    Science.gov (United States)

    Kahlow, Barbara Stadler; Nery, Rodrigo Araldi; Skare, Thelma L; Ribas, Carmen Australia Paredes Marcondes; Ramos, Gabriela Piovezani; Petisco, Roberta Dombroski

    2016-03-01

    Mannose binding lectin is a lectin instrumental in the innate immunity. It recognizes carbohydrate patterns found on the surface of a large number of pathogenic micro-organisms, activating the complement system. However, this protein seems to increase the tissue damage after ischemia. In this paper is reviewed some aspects of harmful role of the mannose binding lectin in ischemia/reperfusion injury. PMID:27120743

  13. Studies of recombinant forms of Aleuria aurantia lectin

    OpenAIRE

    Olausson, Johan

    2009-01-01

    The presented work describes construction and analysis of recombinantly produced forms of Aleuria aurantia lectin (AAL). The binding properties of the produced AAL forms were studied using techniques such as tryptophan fluorescence, hemagglutination analysis, ELISA and surface plasmon resonance analysis. Lectins are proteins that are ubiquitous in nature with the ability to bind specifically to different types of carbohydrates. The physiological function of different lectins is not always kno...

  14. Fine carbohydrate recognition of Euphorbia milii lectin.

    Science.gov (United States)

    Irazoqui, Fernando J; Vozari-Hampe, Magdolna M; Lardone, Ricardo D; Villarreal, Marcos A; Sendra, Victor G; Montich, Guillermo G; Trindade, Vera M; Clausen, Henrik; Nores, Gustavo A

    2005-10-14

    Glycans are key structures involved in biological processes such as cell attachment, migration, and invasion. Information coded on cell-surface glycans is frequently deciphered by proteins, as lectins, that recognize specific carbohydrate topology. Here, we describe the fine carbohydrate specificity of Euphorbia milii lectin (EML). Competitive assays using various sugars showed that GalNAc was the strongest inhibitor, and that the hydroxyl axial position of C4 and acetamido on C2 of GalNAc are critical points of EML recognition. A hydrophobic locus adjacent to GalNAc is also an important region for EML binding. Direct binding assays of EML revealed a stereochemical requirement for a structure adjacent to terminal GalNAc, showing that GalNAc residue is a necessary but not sufficient condition for EML interaction. The capacity of EML to bind epithelial tumor cells makes it a potentially useful tool for study of some over-expressed GalNAc glycoconjugates. PMID:16122701

  15. Lectins discriminate between pathogenic and nonpathogenic South American trypanosomes

    Energy Technology Data Exchange (ETDEWEB)

    de Miranda Santos, I.K.; Pereira, M.E.

    1984-09-01

    Cell surface carbohydrates of Trypanosoma cruzi, Trypanosoma rangeli, and Trypanosoma conorhini were analyzed by a micro-agglutination assay employing 27 highly purified lectins and by binding assays using various /sup 125/I-labeled lectins. The following seven lectins discriminated between the trypanosomes: 1) tomato lectin (an N-acetyl-D-glucosamine-binding protein), both in purified form and as crude tomato juice; 2) Bauhinea purpurea and Sophora japonica lectins (both N-acetyl-D-galactosamine-binding proteins), which selectively agglutinated T. cruzi; 3) Vicia villosa (an N-acetyl-D-galactosamine-binding protein) which was specific for T. rangeli; 4) peanut lectin (a D-galactose-binding protein) both in purified form and as crude saline extract; and 5) Ulex europaeus and Lotus tetragonolobus (both L-fucose-binding proteins) lectins which reacted only with T. conorhini. Binding studies with 125I-labeled lectins were performed to find whether unagglutinated cells of the three different species of trypanosomes might have receptors for these lectins, in which case absence of agglutination could be due to a peculiar arrangement of the receptors. These assays essentially confirmed the agglutination experiments.

  16. Are Vicilins Another Major Class of Legume Lectins?

    Directory of Open Access Journals (Sweden)

    Ana C. Ribeiro

    2014-12-01

    Full Text Available Legume lectins comprise a structurally related, Ca/Mn-dependent, widespread, abundant and well characterized lectin family when compared to the large number of lectins from other sources described in the literature. Strangely enough, no specific function has been assigned to them aside from a possible role in storage and/or defense. Using a recent and fine-tuned methodology capable of specific lectin identification, β-conglutin, Vicia faba vicilin and β-lathyrin, the vicilin storage globulins from Lupinus albus, V. faba and Lathyrus sativus, respectively, were shown to be capable of affinity binding to thoroughly washed erythrocyte membranes and of specific elution with appropriate sugars. Based on this evidence and on sparse data published in the literature, a second family of legume lectins is proposed: the 7S family of storage proteins from leguminous seeds, or family II of legume lectins. These lectins are also structurally related, widespread and well characterized. In addition, they self-aggregate in a Ca/Mg, electrostatic dependent manner and are even more abundant than the family I of legume lectins. Using the same evidence, reserve and defense roles may be attributed to family II of legume lectins.

  17. Lectins discriminate between pathogenic and nonpathogenic South American trypanosomes

    International Nuclear Information System (INIS)

    Cell surface carbohydrates of Trypanosoma cruzi, Trypanosoma rangeli, and Trypanosoma conorhini were analyzed by a micro-agglutination assay employing 27 highly purified lectins and by binding assays using various 125I-labeled lectins. The following seven lectins discriminated between the trypanosomes: 1) tomato lectin (an N-acetyl-D-glucosamine-binding protein), both in purified form and as crude tomato juice; 2) Bauhinea purpurea and Sophora japonica lectins (both N-acetyl-D-galactosamine-binding proteins), which selectively agglutinated T. cruzi; 3) Vicia villosa (an N-acetyl-D-galactosamine-binding protein) which was specific for T. rangeli; 4) peanut lectin (a D-galactose-binding protein) both in purified form and as crude saline extract; and 5) Ulex europaeus and Lotus tetragonolobus (both L-fucose-binding proteins) lectins which reacted only with T. conorhini. Binding studies with 125I-labeled lectins were performed to find whether unagglutinated cells of the three different species of trypanosomes might have receptors for these lectins, in which case absence of agglutination could be due to a peculiar arrangement of the receptors. These assays essentially confirmed the agglutination experiments

  18. Concept, strategy and realization of lectin-based glycan profiling.

    Science.gov (United States)

    Hirabayashi, Jun

    2008-08-01

    Lectins are a diverse group of carbohydrate-binding proteins. Each lectin has its own specificity profile. It is believed that lectins exist in all living organisms that produce glycans. From a practical viewpoint, lectins have been used extensively in biochemical fields including proteomics due to their usefulness as detection and enrichment tools for specific glycans. Nevertheless, they have often been underestimated as probes, especially compared with antibodies, because of their low affinity and broad specificity. However, together with the concept of glycomics, such properties of lectins are now considered to be suitable for the task of 'profiling' in order to cover a wider range of ligands. Recently there has been rapid movement in the field of proteomics aimed at the investigation of glycan-related biomarkers. This is partly because of limitations of the present approach of simply following changes in protein-level expression, without paying sufficient attention to the fact and effects of glycosylation. The trend is reflected in the frequent use of lectins in the contexts of glycoprotein enrichment and glycan profiling. However, there are many aspects to be considered in using lectins, which differ considerably from antibodies. In this article, the author, as a developer of two unique methodologies, frontal affinity chromatography (FAC) and the lectin microarray, describes critical points concerning the use of lectins, together with the concept, strategy and means to achieve advances in these emerging glycan profiling technologies. PMID:18390573

  19. C-type natriuretic peptide in prostate cancer

    DEFF Research Database (Denmark)

    Nielsen, Soeren Junge; Iversen, Peter; Rehfeld, Jens F.;

    2009-01-01

    C-type natriuretic peptide (CNP) is expressed in the male reproductive organs in pigs. To examine whether the human prostate also expresses the CNP gene, we measured CNP and N-terminal proCNP in prostate cancer tissue extracts and performed immunohistochemical biopsy staining. Additionally, pro......CNP-derived peptides were quantitated in plasma from patients with prostate cancer. Blood was collected from healthy controls and patients before surgery for localized prostate cancer. Tissue extracts were prepared from tissue biopsies obtained from radical prostatectomy surgery. N-terminal proCNP, proCNP (1-50) and...... CNP were measured in plasma and tissue extracts. Biopsies were stained for CNP-22 and N-terminal proCNP. Tissue extracts from human prostate cancer contained mostly N-terminal proCNP [median 5.3 pmol/g tissue (range 1.0-12.9)] and less CNP [0.14 pmol/g tissue (0.01-1.34)]. Immunohistochemistry...

  20. C-Type Natriuretic Peptide Analog as Therapy for Achondroplasia.

    Science.gov (United States)

    Legeai-Mallet, Laurence

    2016-01-01

    Fibroblast growth factor receptor 3 (FGFR3) is an important regulator of bone formation. Gain-of-function mutations in the FGFR3 gene result in chondrodysplasias which include achondroplasia (ACH), the most common form of dwarfism, in which skull, appendicular and axial skeletons are affected. The skeletal phenotype of patients with ACH showed defective proliferation and differentiation of the chondrocytes in the growth plate cartilage. Both endochondral and membranous ossification processes are disrupted during development. At cellular level, Fgfr3 mutations induce increased phosphorylation of the tyrosine kinase receptor FGFR3, which correlate with an enhanced activation of its downstream signaling pathways. Potential therapeutic strategies have emerged for ACH. Several preclinical studies have been conducted such as the C-type natriuretic peptide (CNP) analog (BMN111), intermittent parathyroid hormone injections, soluble FGFR3 therapy, and meclozine and statin treatments. Among the putative targets to antagonize FGFR3 signaling, CNP (or BMN111) is one of the most promising strategies. BMN111 acts as a key regulator of longitudinal bone growth by downregulating the mitogen-activated protein kinase pathway, which is activated as a result of a FGFR3 gain-of-function mutation. Preclinical studies showed that BMN111 treatment led to a large improvement in skeletal parameters in Fgfr3Y367C/+ mice mimicking ACH. In 2014, a clinical trial (phase 2) of BMN111 in pediatric patients with ACH has started. This first clinical trial marks the first big step towards real treatment for these patients. PMID:26684019

  1. Mathematical model of various statements of C-type Language

    Directory of Open Access Journals (Sweden)

    Manoj Kumar Srivastav

    2013-12-01

    Full Text Available Some of the important components of high level languages are statements, keywords, variable declarations, arrays, user defined functions etc. In case of object oriented programming language we use class, object, inheritance, operator overloading, function overloading, polymorphism etc. There are some common category of statements such as control statement, loop statements etc. Pointers are also one important concept in C-language. User defined functions, function subprograms or subroutines are also important concepts in different programming languages. The language like ALGOL was developed using Chomsky context free grammar. The similar concept used in C-type languages. The high level languages are now based on mathematical derivations and logic. Most of the components of any high level language can be obtained from simple mathematical logic and derivations. In the present study the authors have tried to give some unified mathematical model of few statements, arrays, user defined functions of C-language. However, the present method may further be extended to any other high level language.

  2. High-level expression and characterization of a glycosylated human cementum protein 1 with lectin activity.

    Science.gov (United States)

    Romo-Arévalo, Enrique; Arzate, Higinio; Montoya-Ayala, Gonzalo; Rodríguez-Romero, Adela

    2016-01-01

    This work aims to contribute to the knowledge of human cementum protein 1 (CEMP1), its conformational characteristics and influence during the biomineralization process. The results revealed that hrCEMP1 expressed in Pichia pastoris is a 2.4% glycosylated, thermostable protein which possesses a molecular mass of 28 770 Da. The circular dichroism spectrum indicated a secondary structure content of 28.6% of alpha-helix, 9.9% of beta-sheet and 61.5% of random-coil forms. Biological activity assays demonstrated that hrCEMP1 nucleates and regulates hydroxyapatite crystal growth. Hereby, it is demonstrated for the first time that CEMP1 has a (C-type) lectin-like activity and specifically recognizes mannopyranoside. The information produced by this biochemical and structural characterization may contribute to understand more fully the biological functions of CEMP1. PMID:26763148

  3. Pancreatitis-associated protein: From a lectin to an anti-inflammatory cytokine

    Institute of Scientific and Technical Information of China (English)

    Daniel Closa; Yoshiharu Motoo; Juan L Iovanna

    2007-01-01

    Pancreatitis-associated protein (PAP) was discovered in the pancreatic juice of rats with acute pancreatitis. PAP is a 16 kDa secretory protein structurally related to the C-type lectins although classical lectin-related function has not been reported yet. Then, it was demonstrated that PAP expression may be activated in some tissues in a constitutive or injury- and inflammation-induced manner. More recently, it has been found that PAP acts as an anti-inflammatory factor in vitro and in vivo.PAP expression can be induced by several pro- and anti-inflammatory cytokines and by itself through a JAK/STAT3-dependent pathway. PAP is able to activate the expression of the anti-inflammatory factor SOCS3 through the JAK/STAT3-dependent pathway. The JAK/STAT3/SOCS3 pathway seems to be a common point between PAP and several cytokines. Therefore,it is reasonable to propose that PAP is a new antiinflammatory cytokine.

  4. Mannan-binding lectin activates C3 and the

    DEFF Research Database (Denmark)

    Selander, B.; Martensson, U.; Weintraub, A.;

    2006-01-01

    Lectin pathway activation of C3 is known to involve target recognition by mannan-binding lectin (MBL) or ficolins and generation of classical pathway C3 convertase via cleavage of C4 and C2 by MBL-associated serine protease 2 (MASP-2). We investigated C3 activation in C2-deficient human sera and in...

  5. Assessment of lectin inactivation by heat and digestion.

    Science.gov (United States)

    Pusztai, A; Grant, G

    1998-01-01

    Proteins/glycoproteins from plants, particularly lectins, are more resistant to heat denaturation than animal proteins (1, 2). With legume seeds, whose lectin content is appreciable, this presents potentially serious problems in nutritional practice. Therefore, before they can be used safely, legume-based food/ feeds usually require thorough and expensive heat processing to inactivate antinutritive components. Indeed, dry or moist heating of seeds at 70°C for several h has little or no effect on their lectin activity (Fig. 1) and treatment at much higher temperatures is needed to inactivate the biological and antinutritional effects of legume lectins (1, 2). The safety aspect is even more serious with some monocot lectins, such as wheatgerm agglutinin or a number of oilseed lectins, such as peanut agglutinin and many others because they are extremely heat stable and normal cooking or other conventional heat treatments may fail to inactivate them (3) Thus, the best way to avoid potential harmful effects of these heat-resistant lectins is to limit their dietary intake to a minimum. Fig. 1. Loss of lectin activity during aqueous heat treatment of soybean at various temperatures. PMID:21374488

  6. Mannose-binding lectin genetics: from A to Z

    DEFF Research Database (Denmark)

    Garred, Peter

    2008-01-01

    MBL (mannose-binding lectin) is primarily a liver-derived collagen-like serum protein. It binds sugar structures on micro-organisms and on dying host cells and is one of the four known mediators that initiate activation of the complement system via the lectin pathway. Common variant alleles...

  7. Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential

    Directory of Open Access Journals (Sweden)

    Johan Gardères

    2015-08-01

    Full Text Available An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial systems. The characterization of the lectins was performed by Edman degradation or mass spectrometry. Regarding their physiological roles, sponge lectins showed to be involved in morphogenesis and cell interaction, biomineralization and spiculogenesis, as well as host defense mechanisms and potentially in the association between the sponge and its microorganisms. In addition, these lectins exhibited a broad range of bioactivities, including modulation of inflammatory response, antimicrobial and cytotoxic activities, as well as anticancer and neuromodulatory activity. In view of their potential pharmacological applications, sponge lectins constitute promising molecules of biotechnological interest.

  8. Porifera Lectins: Diversity, Physiological Roles and Biotechnological Potential.

    Science.gov (United States)

    Gardères, Johan; Bourguet-Kondracki, Marie-Lise; Hamer, Bojan; Batel, Renato; Schröder, Heinz C; Müller, Werner E G

    2015-08-01

    An overview on the diversity of 39 lectins from the phylum Porifera is presented, including 38 lectins, which were identified from the class of demosponges, and one lectin from the class of hexactinellida. Their purification from crude extracts was mainly performed by using affinity chromatography and gel filtration techniques. Other protocols were also developed in order to collect and study sponge lectins, including screening of sponge genomes and expression in heterologous bacterial systems. The characterization of the lectins was performed by Edman degradation or mass spectrometry. Regarding their physiological roles, sponge lectins showed to be involved in morphogenesis and cell interaction, biomineralization and spiculogenesis, as well as host defense mechanisms and potentially in the association between the sponge and its microorganisms. In addition, these lectins exhibited a broad range of bioactivities, including modulation of inflammatory response, antimicrobial and cytotoxic activities, as well as anticancer and neuromodulatory activity. In view of their potential pharmacological applications, sponge lectins constitute promising molecules of biotechnological interest. PMID:26262628

  9. Studies of the articular cartilage proteoglycan aggrecan in health and osteoarthritis. Evidence for molecular heterogeneity and extensive molecular changes in disease.

    Science.gov (United States)

    Rizkalla, G; Reiner, A; Bogoch, E; Poole, A R

    1992-01-01

    Changes in the structure of the proteoglycan aggrecan (PG) of articular cartilage were determined immunochemically by RIA and gel chromatography and related to cartilage degeneration documented histologically by the Mankin grading system. Monoclonal antibodies to glycosaminoglycan epitopes were used. In all cartilages, three chondroitin sulfate (CS)-rich populations of large size were observed in addition to a smaller keratan sulfate (KS)-rich population. In grades 7-13 OA cartilages (phase II), molecules were significantly larger than the equivalent molecules of grades 2-6 (phase I). CS chain lengths remained unchanged. In most OA cartilages, a CS epitope 846 was elevated in content, this being most marked in phase II (mean: fivefold). Loss of uronic acid, KS, and hyaluronic acid were only pronounced in phase II OA because of variations in normal contents. Aggregation of PG was unchanged (50-60%) or reduced in OA cartilages, but molecules bearing epitope 846 exhibited almost complete aggregation in normal cartilages. This study provides evidence for the capacity of OA cartilage to synthesize new aggrecan molecules to replace those damaged and lost by disease-related changes. It also defines two phases of PG change in OA: an early predominantly degenerate phase I followed by a net reparative phase II accompanied by net loss of these molecules. Images PMID:1281828

  10. Nutritional evaluation of lectin-free soybeans for poultry.

    Science.gov (United States)

    Douglas, M W; Parsons, C M; Hymowitz, T

    1999-01-01

    This study evaluated the nutritional value of raw lectin-free soybeans in comparison with raw Kunitz trypsin inhibitor-free soybeans, raw conventional soybeans, and commercial heat processed soybean meal (SBM). Analyzed lectin values (milligrams per kilogram) were 7.2, 7.1, and lectin-free soybeans, respectively. Three experiments were conducted using New Hampshire x Columbian male chicks fed 23% CP dextrose-soybean diets from 8 to 17 d of age. Growth performance of chicks fed lectin-free soybeans was greater (P lectin-free soybeans was lower than that of chicks fed Kunitz-free soybeans or SBM. The SBM yielded weight gains and feed efficiencies that were much higher than those observed from any of the raw soybeans. True amino acid digestibility and TMEn of the lectin-free and conventional soybeans were determined using the precision-fed cecectomized rooster assay. Seven roosters were crop-intubated with 30 g of soybeans and excreta were collected for 48 h. Digestibility coefficients of most amino acids for lectin-free soybeans were 5 to 8 percentage units higher than those for conventional soybeans, but the differences were not significant (P > 0.05). Likewise, the TMEn for lectin-free soybeans was 11% higher than that for raw conventional soybeans (3.577 vs 3.227 kcal/g DM) but the difference was not significant (P > 0.05). The results of this study indicate that the nutritional value of raw lectin-free soybeans is greater than raw conventional soybeans but is less than raw Kunitz-free soybeans and SBM, suggesting that trypsin inhibitor is a greater antinutritional factor than lectins. PMID:10023754

  11. Lectin activity in mycelial extracts of Fusarium species.

    Science.gov (United States)

    Bhari, Ranjeeta; Kaur, Bhawanpreet; Singh, Ram S

    2016-01-01

    Lectins are non-immunogenic carbohydrate-recognizing proteins that bind to glycoproteins, glycolipids, or polysaccharides with high affinity and exhibit remarkable ability to agglutinate erythrocytes and other cells. In the present study, ten Fusarium species previously not explored for lectins were screened for the presence of lectin activity. Mycelial extracts of F. fujikuroi, F. beomiformii, F. begoniae, F. nisikadoi, F. anthophilum, F. incarnatum, and F. tabacinum manifested agglutination of rabbit erythrocytes. Neuraminidase treatment of rabbit erythrocytes increased lectin titers of F. nisikadoi and F. tabacinum extracts, whereas the protease treatment resulted in a significant decline in agglutination by most of the lectins. Results of hapten inhibition studies demonstrated unique carbohydrate specificity of Fusarium lectins toward O-acetyl sialic acids. Activity of the majority of Fusarium lectins exhibited binding affinity to d-ribose, l-fucose, d-glucose, l-arabinose, d-mannitol, d-galactosamine hydrochloride, d-galacturonic acid, N-acetyl-d-galactosamine, N-acetyl-neuraminic acid, 2-deoxy-d-ribose, fetuin, asialofetuin, and bovine submaxillary mucin. Melibiose and N-glycolyl neuraminic acid did not inhibit the activity of any of the Fusarium lectins. Mycelial extracts of F. begoniae, F. nisikadoi, F. anthophilum, and F. incarnatum interacted with most of the carbohydrates tested. F. fujikuroi and F. anthophilum extracts displayed strong interaction with starch. The expression of lectin activity as a function of culture age was investigated. Most species displayed lectin activity on the 7th day of cultivation, and it varied with progressing of culture age. PMID:27237111

  12. Genetic influences on mannan-binding lectin (MBL) and mannan-binding lectin associated serine protease-2 (MASP-2) activity

    DEFF Research Database (Denmark)

    Sørensen, Grith Lykke; Petersen, Inge; Thiel, Steffen; Fenger, Mogens; Christensen, Kaare; Kyvik, Kirsten O; Sørensen, Thorkild I A; Holmskov, Uffe; Jensenius, Jens Christian

    2007-01-01

    The lectin pathway of the complement system is activated when Mannan-binding lectin (MBL) in complex with MASP-2 binds microorganisms. Polymorphisms in both genes are responsible for low serum levels, which associate with increased risk of infection and autoimmune disease. The present study...

  13. Tick lectins and fibrinogen-related proteins

    Czech Academy of Sciences Publication Activity Database

    Grubhoffer, Libor; Rego, Ryan O. M.; Hajdušek, Ondřej; Hypša, Václav; Kovář, Vojtěch; Rudenko, Natalia; Oliver, J. H., Jr.

    Cambridge : Cambridge University Press, 2008 - (Bowman, A.; Nuttall, P.), s. 127-142 ISBN 978-0-521-86761-0 R&D Projects: GA MŠk(CZ) LC06009; GA ČR(CZ) GA524/06/1479; GA AV ČR IAA6022001; GA ČR(CZ) GA206/04/0520 Grant ostatní: National Institutes of Health(US) R37 AI-24899 Institutional research plan: CEZ:AV0Z60220518 Keywords : lectin * fibrinogen-related protein * protein-saccharide interaction Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine

  14. Noncovalent PEGylation via Lectin-Glycopolymer Interactions.

    Science.gov (United States)

    Antonik, Paweł M; Eissa, Ahmed M; Round, Adam R; Cameron, Neil R; Crowley, Peter B

    2016-08-01

    PEGylation, the covalent modification of proteins with polyethylene glycol, is an abundantly used technique to improve the pharmacokinetics of therapeutic proteins. The drawback with this methodology is that the covalently attached PEG can impede the biological activity (e.g., reduced receptor-binding capacity). Protein therapeutics with "disposable" PEG modifiers have potential advantages over the current technology. Here, we show that a protein-polymer "Medusa complex" is formed by the combination of a hexavalent lectin with a glycopolymer. Using NMR spectroscopy, small-angle X-ray scattering (SAXS), size exclusion chromatography, and native gel electrophoresis it was demonstrated that the fucose-binding lectin RSL and a fucose-capped polyethylene glycol (Fuc-PEG) form a multimeric assembly. All of the experimental methods provided evidence of noncovalent PEGylation with a concomitant increase in molecular mass and hydrodynamic radius. The affinity of the protein-polymer complex was determined by ITC and competition experiments to be in the micromolar range, suggesting that such systems have potential biomedical applications. PMID:27403588

  15. Histological and Lectin Histochemical Studies on the Olfactory and Respiratory Mucosae of the Sheep

    OpenAIRE

    Ibrahim, Dalia; NAKAMUTA, Nobuaki; TANIGUCHI, Kazumi; Yamamoto, Yoshio; TANIGUCHI, Kazuyuki

    2013-01-01

    ABSTRACT The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-whea...

  16. Putative glycoprotein and glycolipid polymorphonuclear leukocyte receptors for the Actinomyces naeslundii WVU45 fimbrial lectin.

    OpenAIRE

    Sandberg, A L; Ruhl, S; Joralmon, R A; Brennan, M J; Sutphin, M J; Cisar, J O

    1995-01-01

    Recognition of receptors on sialidase-treated polymorphonuclear leukocytes (PMNs) by the Gal/GalNAc lectin associated with the type 2 fimbriae of certain strains of actinomyces results in activation of the PMNs, phagocytosis, and destruction of the bacteria. In the present study, plant lectins were utilized as probes to identify putative PMN receptors for the actinomyces lectin. The Gal-reactive lectin from Ricinus communis (RCAI), the Gal/GalNAc-reactive lectins from R. communis (RCAII) and ...

  17. Chicken C-type lectin-like receptor B-NK, expressed on NK and T cell subsets, binds to a ligand on activated splenocytes

    Czech Academy of Sciences Publication Activity Database

    Viertiboeck, B.C.; Wortmann, A.; Schmitt, R.; Plachý, Jiří; Gobel, T.W.

    2008-01-01

    Roč. 45, č. 5 (2008), s. 1398-1404. ISSN 0161-5890 Institutional research plan: CEZ:AV0Z50520514 Keywords : Chicken NK cell receptor Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.555, year: 2008

  18. Potential immunomodulatory effects of plant lectins in Schistosoma mansoni infection.

    Science.gov (United States)

    Reis, Eliana A G; Athanazio, Daniel A; Cavada, Benildo Sousa; Teixeira, Edson Holanda; de Paulo Teixeira Pinto, Vicente; Carmo, Theomira M A; Reis, Alice; Trocolli, Graziela; Croda, Julio; Harn, Donald; Barral-Netto, Manoel; Reis, Mitermayer G

    2008-01-01

    Lectins are sugar-binding glycoproteins that can stimulate, in a non-antigen-specific fashion, lymphocytes, leading to proliferation and cytokine production. Some lectins are utilized as in vitro mitogenic lymphocyte stimulators and their use as immunomodulators against infectious diseases has been evaluated experimentally. In the experimental murine model, the immune response to schistosomiasis is Th1-like during the initial stage of infection, with a shift towards a Th2-like response after oviposition. We report the response of schistosomiasis patients' (n=37) peripheral blood mononuclear cells (PBMC) to stimulation by lectins, including newly isolated lectins from Brazilian flora, and by Schistosomamansoni soluble egg antigens (SEA). Cytokine production upon lectin stimulation ex vivo was assessed in PBMC supernatants, collected at 24 and 72 h, by sandwich ELISA to IL-5, IL-10, TNF-alpha and IFN-gamma. In PBMC from infected patients all but one of the lectins induced a Th2-like cytokine response, characterized by elevated IL-5 production that was higher than that induced by SEA stimulation alone. Our results show that the Th2 environment present during schistosomiasis is not affected and that it may be further stimulated by the presence of lectins. PMID:18579103

  19. Effects of lectin ingestion on animal growth and internal organs.

    Science.gov (United States)

    Pusztai, A

    1998-01-01

    Lectins are essential and omnipresent plant constituents. As many foods are of plant origin, the daily ingestion of lectins by both humans and animals is appreciable. For example, in an ad hoc survey, 53 edible plants were shown to contain lectins and approx 30% of fresh and processed food regularly consumed by humans had significant hemagglutinating activity (1). The situation is potentially even more acute in animal nutrition because animal diet is less diverse than that of humans, and in most instances foodstuffs are not thoroughly heat-treated. This is particularly significant in the light of our finding a correlation between lectin activity and antinutritional effects (2). As in evolution, the mammalian gut has been regularly exposed to lectins, they must have played an important part in the development of the digestive system. Although based on experience, most overtly toxic plants have been eliminated from the diet, many plants with appreciable lectin content are still consumed because it has not been easy to relate growth retardation and antinutritional, mild allergic or other subclinical symptoms to the food consumed or a particular component of it. As some lectins are at least partially heat stable and most survive the passage through the gut in functionally and immunologically intact form, their interaction with the gut surface epithelium (3) can damage the gut at high dietary intakes and this may lead to digestive disorders/diseases in some instances. However, it is not generally appreciated that not all lectins are antinutrients and indeed some may have beneficial effects and be of potential value in nutritional practice. Accordingly, it is of considerable importance to establish whether a lectin has deleterious or potentially beneficial effects for mammals. Unfortunately at present there are no adequate in vitro methods to do this reliably and it is usually necessary to carry out in vivo animal feeding studies, despite their relatively cumbersome

  20. Lectin immuno tests: quantitation and titration of antigens and antibodies using lectin-antibody conjugates

    International Nuclear Information System (INIS)

    The authors have investigated the possibility of using lectin-antibody conjugates as general reagents in immunological procedures requiring a labeled antigen or antibody. Using these conjugates, labeling is achieved through saccharide binding sites of lectins which operate as acceptors for glycoconjugate marker substances added secondarily. Marker substances used in this work were enzymes, radioactively labeled glycoconjugates and erythrocytes, but other markers can also be used. Using the first two markers, antigens and antibodies were determined with accuracy and sensitivity equal to those of conventional enzyme or radioimmunoassays. Using erythrocytes as a marker, a simple erythro-adsorption procedure, possibly followed by hemolysis, has been developed which allowed the titration of antigens and antibodies to be carried out with a sensitivity at least equal to enzyme or radioimmunoassays. (Auth.)

  1. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    DEFF Research Database (Denmark)

    Wang, Bijue; Chen, Pingping; Jensen, Anne-Christine Bay; Karsdal, Morten A; Madsen, Suzi H; Sondergaard, Bodil-Cecilie; Zheng, Qinlong; Qvist, Per

    2009-01-01

    . We found that (1) aggrecanase-derived aggrecan fragments are released in the early (day 2-7) and mid phase (day 9-14) into the supernatant from bovine explants cultures stimulated with catabolic cytokines, (2) the release of NITEGE(373 )neo-epitopes are delayed compared to the corresponding (374......BACKGROUND: Progressive loss of articular cartilage is a central hallmark in many joint disease, however, the relative importance of individual proteolytic pathways leading to cartilage erosion is at present unknown. We therefore investigated the time-dependant release ex vivo of MMP- and...... cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM) and tumor necrosis factor alpha (TNFalpha). In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo...

  2. Structure and Function of Mammalian Carbohydrate-Lectin Interactions

    Science.gov (United States)

    Anderson, Kevin; Evers, David; Rice, Kevin G.

    Over the past three decades the field of glycobiology has expanded beyond a basic understanding of the structure and biosynthesis of glycoprotein, proteoglycans, and glycolipids toward a more detailed picture of how these molecules afford communication through binding to mammalian lectins. Although the number of different mammalian lectin domains appears to be finite and even much smaller than early estimates predicated based on the diversity of glycan structures, nature appears capable of using these in numerous combinations to fine tune specificity. The following provides an overview of the major classes of mammalian lectins and discusses their glycan binding specificity. The review provides a snapshot of the field of glycobiology that continues to grow providing an increasing number of examples of biological processes that rely upon glycan-lectin binding.

  3. Lectins stain cells differentially in the coral, Montipora capitata

    Science.gov (United States)

    Work, Thierry M.; Farah, Yael

    2014-01-01

    A limitation in our understanding of coral disease pathology and cellular pathogenesis is a lack of reagents to characterize coral cells. We evaluated the utility of plant lectins to stain tissues of a dominant coral, Montipora capitata, from Hawaii. Of 22 lectins evaluated, nine of these stained structures in the upper or basal body wall of corals. Specific structures revealed by lectins that were not considered distinct or evident on routine hematoxylin and eosin sections of coral tissues included apical and basal granules in gastrodermis and epidermis, cnidoglandular tract and actinopharynx cell surface membranes, capsules of mature holotrichous isorhizas, and perivitelline and periseminal cells. Plant lectins could prove useful to further our understanding of coral physiology, anatomy, cell biology, and disease pathogenesis.

  4. Detection of Lectins by Saccharide-gold Nanoparticle Conjugates

    Institute of Scientific and Technical Information of China (English)

    HOU Qiong; WANG Jin-e; LIU Xia; LI Xiao-kun; MA Li-na; DUAN Wu-biao; WANG Zhen-xin

    2012-01-01

    A general functionalization strategy was reported,which enables one to conjugate saccharide(SA) on gold nanoparticle(GNP) surface without affecting SA properties.First,disulfide phenylboronic acid(Bor) functionalized GNPs(Bor@GNPs) were synthesized by the reaction of citrate stabilized GNPs of 13 nm in diameter with the mixture of Bor and pentapeptide(Cys-Ala-Leu-Asn-Asn,CALNN).Subsequently,the SA-GNP conjugates(SA@GNPs) were prepared by coupling SA to the GNP surface via the reaction of phenylboronic acid(PBA) with the cis-diol configuration in SA.The interactions of three SA@GNPs with three lectins have been analyzed by UV-visible spectroscopic and transmission electronic microscopic(TEM) techniques,respectively.The experimental results demonstrate that SA@GNPs can efficiently bind to lectins and show a great promise as optical probes for monitoring specific affinities of lectins for SA,and detecting lectins with high sensitivity.

  5. Protozoa lectins and their role in host-pathogen interactions.

    Science.gov (United States)

    Singh, Ram Sarup; Walia, Amandeep Kaur; Kanwar, Jagat Rakesh

    2016-01-01

    Lectins are proteins/glycoproteins of non-immune origin that agglutinate red blood cells, lymphocytes, fibroblasts, etc., and bind reversibly to carbohydrates present on the apposing cells. They have at least two carbohydrate binding sites and their binding can be inhibited by one or more carbohydrates. Owing to carbohydrate binding specificity of lectins, they mediate cell-cell interactions and play role in protozoan adhesion and host cell cytotoxicity, thus are central to the pathogenic property of the parasite. Several parasitic protozoa possess lectins which mediate parasite adherence to host cells based on their carbohydrate specificities. These interactions could be exploited for development of novel therapeutics, targeting the adherence and thus helpful in eradicating wide spread of protozoan diseases. The current review highlights the present state knowledge with regard to protozoal lectins with an emphasis on their haemagglutination activity, carbohydrate specificity, characteristics and also their role in pathogenesis notably as adhesion molecules, thereby aiding the pathogen in disease establishment. PMID:27268207

  6. Insights into Animal and Plant Lectins with Antimicrobial Activities

    Directory of Open Access Journals (Sweden)

    Renata de Oliveira Dias

    2015-01-01

    Full Text Available Lectins are multivalent proteins with the ability to recognize and bind diverse carbohydrate structures. The glyco -binding and diverse molecular structures observed in these protein classes make them a large and heterogeneous group with a wide range of biological activities in microorganisms, animals and plants. Lectins from plants and animals are commonly used in direct defense against pathogens and in immune regulation. This review focuses on sources of animal and plant lectins, describing their functional classification and tridimensional structures, relating these properties with biotechnological purposes, including antimicrobial activities. In summary, this work focuses on structural-functional elucidation of diverse lectin groups, shedding some light on host-pathogen interactions; it also examines their emergence as biotechnological tools through gene manipulation and development of new drugs.

  7. Sweet complementarity: the functional pairing of glycans with lectins.

    Science.gov (United States)

    Gabius, H-J; Manning, J C; Kopitz, J; André, S; Kaltner, H

    2016-05-01

    Carbohydrates establish the third alphabet of life. As part of cellular glycoconjugates, the glycans generate a multitude of signals in a minimum of space. The presence of distinct glycotopes and the glycome diversity are mapped by sugar receptors (antibodies and lectins). Endogenous (tissue) lectins can read the sugar-encoded information and translate it into functional aspects of cell sociology. Illustrated by instructive examples, each glycan has its own ligand properties. Lectins with different folds can converge to target the same epitope, while intrafamily diversification enables functional cooperation and antagonism. The emerging evidence for the concept of a network calls for a detailed fingerprinting. Due to the high degree of plasticity and dynamics of the display of genes for lectins the validity of extrapolations between different organisms of the phylogenetic tree yet is inevitably limited. PMID:26956894

  8. Identification of lectin-binding proteins in Chlamydia species.

    OpenAIRE

    Swanson, A F; Kuo, C. C.

    1990-01-01

    Lectin-binding proteins of chlamydiae were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. All three Chlamydia species tested expressed two proteins when whole-elementary-body lysates were reacted with the biotinylated lectin Dolichos biflorus agglutinin. The protein with a molecular mass of 18 kilodaltons (kDa) responded strongly compared with a higher-molecular-mass protein that varied from 27 to 32 kDa with each chlamydia strain tested. Among six l...

  9. Antibacterial activity of a lectin-like Burkholderia cenocepacia protein

    OpenAIRE

    Ghequire, Maarten; De Canck, Evelien; Wattiau, Pierre; Van Winge, Iris; Loris, Remy; Coenye, Tom; De Mot, René

    2013-01-01

    Abstract Bacteriocins of the LlpA family have previously been characterized in the γ-proteobacteria Pseudomonas and Xanthomonas. These proteins are composed of two MMBL (monocot mannose-binding lectin) domains, a module predominantly and abundantly found in lectins from monocot plants. Genes encoding four different types of LlpA-like proteins were identified in genomes from strains belonging to the Burkholderia cepacia complex (Bcc) and the Burkholderia pseudomallei group. A selected recombin...

  10. Evaluation of leguminous lectins activities against bacterial biofilm formation

    OpenAIRE

    Carneiro, Victor Alves; Cavalcante, Theodora Thays Arruda; Teixeira, Edson Holanda; Cavada, Benildo Sousa; Oliveira, Rosário; Henriques, Mariana; Pereira, Maria Olívia

    2010-01-01

    Biofilms are composed by microbial cells that are irreversibly associated with a surface and enclosed in a matrix of polymeric material. Lectins are sugar binding proteins of non immune origin that agglutinate cells and ⁄ or precipitate glycoconjugate molecules. Due to their capacity to bind and recognize specific carbohydrates, lectins can be a potent tool in biofilm studies. The search for potential phytochemicals as anti-biofilm agents has become an active area of research, and these protei...

  11. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

    OpenAIRE

    Renata Angeli; da Paz, Nathalia V. N.; Maciel, Jackeline C.; Flávia F. B. Araújo; Paiva, Patrícia M. G.; Glícia M. T. Calazans; Ana Paula Valente; Almeida, Fábio C. L.; Coelho, Luana C. B. B.; Carvalho, Luiz B; Silva, Maria da Paz C.; Maria Tereza dos Santos Correia

    2009-01-01

    A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A) and Cratylia mollis (Cramoll 1 and Cramoll 1, 4) did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammon...

  12. Antifungal activity of lectins against yeast of vaginal secretion

    Directory of Open Access Journals (Sweden)

    Bruno Severo Gomes

    2012-06-01

    Full Text Available Lectins are carbohydrate-binding proteins of non-imune origin. This group of proteins is distributed widely in nature and they have been found in viruses, microorganisms, plants and animals. Lectins of plants have been isolated and characterized according to their chemical, physical-chemical, structural and biological properties. Among their biological activities, we can stress its fungicidal action. It has been previously described the effect of the lectins Dviol, DRL, ConBr and LSL obtained from the seeds of leguminous plants on the growth of yeasts isolated from vaginal secretions. In the present work the experiments were carried out in microtiter plates and the results interpreted by both methods: visual observations and a microplate reader at 530nm. The lectin concentrations varied from 0.5 to 256µg/mL, and the inoculum was established between 65-70% of trammitance. All yeast samples isolated from vaginal secretion were evaluated taxonomically, where were observed macroscopic and microscopic characteristics to each species. The LSL lectin did not demonstrate any antifungal activity to any isolate studied. The other lectins DRL, ConBr and DvioL, showed antifungal potential against yeast isolated from vaginal secretion. These findings offering offer a promising field of investigation to develop new therapeutic strategies against vaginal yeast infections, collaborating to improve women's health.

  13. Detection and identification of tissue specific lectins of the tsetse fly, Glossina tachinoides: Midgut lectin activity with lipopolysaccharide binding specificity

    International Nuclear Information System (INIS)

    Lectin that agglutinates human and animal red blood cells (RBCs) was demonstrated in midgut extracts of Glossina tachinoides. The highest haemagglutination titres were against pig and rabbit RBCs. Treatment of rabbit RBCs with pronase, trypsin, neuraminidase, bromelain, glutaraldehyde and periodate reduced the agglutination titres. The lectin is specific for amino, methyl and deoxy derivates of glucose, amino and methyl derivates of mannose, D-galactosamine, N-acetylneuraminic acid and trehalose. In addition, very high reactivity against the lipopolysaccharide of E. coli K 235 was found. Lectin is secreted to the midgut lumen. It consists of a 27 kilodalton protein component that is not glycosylated. Sandwich ELISA permits quantification of lectin in tissue samples. (author). 15 refs, 3 figs, 1 tab

  14. Lectin histochemistry of normal and neoplastic peripheral nerve sheath. 2. Lectin binding patterns of schwannoma and neurofibroma.

    Science.gov (United States)

    Matsumura, K; Nakasu, S; Nioka, H; Handa, J

    1993-01-01

    Lectin binding patterns of 31 schwannomas and 6 neurofibromas were examined using 12 lectins, and the results were compared with those of normal peripheral nerves. Tumors obtained from 10 cases of neurofibromatosis and 4 recurrent schwannomas were included. Changes of glycoconjugates were observed in association with a neoplastic transformation of Schwann cells; Arachis hypogaea (PNA) staining after neuraminidase treatment seen in normal Schwann cells was reduced in schwannoma of Antoni type A, and bindings with Glycine max (SBA) and Helix pomatia (HPA) after sialic acid removal, which were not seen in normal Schwann cells, appeared in schwannoma cells. Intensities of staining of tumor cells with each lectin were higher in Antoni type B than those in Antoni type A. No differences in lectin binding patterns were observed between schwannomas in patients with neurofibromatosis or recurrent schwannomas and ordinary, primary schwannomas in patients without stigmata of neurofibromatosis. Lectin binding patterns of Schwann cells and perineurial cells in neurofibroma were almost similar to those in normal peripheral nerves with an exception of faint stain of Schwann cells with HPA after neuraminidase pretreatment. This result suggests differences in extent of differentiation between schwannoma cells and neoplastic Schwann cells in neurofibroma. Specific PNA binding to perineurial cells in neurofibroma indicates the significance of this lectin as a marker of these cells. PMID:8310811

  15. Lectin histochemistry of normal and neoplastic peripheral nerve sheath. 1. Lectin binding pattern of normal peripheral nerve in man.

    Science.gov (United States)

    Matsumura, K; Nakasu, S; Nioka, H; Handa, J

    1993-01-01

    The binding patterns of lectins to normal peripheral nerves were examined. Twelve biotinylated lectins were used in this study; Canavalia ensiformis (Con A), Pisum sativum (PSA), Lens culinaris (LCA), Ricinus communis 1 (RCA-1), Arachis hypogaea (PNA), Glycine max (SBA), Sophora japonica (SJA), Bandeiraea simplicifolia 1 (BSL-1), Triticum vulgaris (WGA), succinylated WGA (s-WGA), Ulex europaeus 1 (UEA-1) and Helix pomatia (HPA). Cytoplasm of Schwann cells and perineurial cells was stained by Con A, PSA, LCA, s-WGA and WGA. PNA showed specific binding to perineurial cells, while after neuraminidase treatment stain with this lectin was demonstrated also in Schwann cells. Myelin sheaths were stained with fewer lectins. SBA and HPA with sialic acid removal rarely showed reactivity to the peripheral nerve structure in surgical specimens, in contrast to clear staining of Schwann cells, perineurial cells and myelin sheaths in autopsy specimens. The present study shows distinct lectin stainings of specific structures of the normal human peripheral nerves, and provides important basic information on the alterations of lectin binding patterns during pathological processes in the peripheral nerves. PMID:8310810

  16. Coloidal gold, ferritin and peroxidase as markers for electron microscopic double labeling lectin techniques.

    Science.gov (United States)

    Roth, J; Binder, M

    1978-03-01

    Three markers, colloidal gold, ferritin and peroxidase, were checked for usefulness in double labeling of lectin-binding sites. The amount of various lectins for the stabilization of good sols of a different particle size was evaluated. Several lectin-gold complexes were prepared for electron microscopic labeling purposes, and the optimal amount of various lectins needed for stabilization of gold solutions of a different particle size was determined. The following combinations were investigated for their usefulness in labeling two different lectin-binding sites: lectin-gold and lectin-gold (different particle size), lectin-gold and lectin-ferritin, as well as lectin-ferritin and lectin-peroxidase. Of these combinations the latter did not give satisfactory results for double labeling. In all single and double labeling techniques with the above mentioned markers the quantitative evaluation of the number of lectin-binding sites is not feasible, but these techniques will be of considerable value for the investigation of the dynamics of different lectin-binding sites on the cell surface. PMID:632554

  17. Specificity analysis of lectins and antibodies using remodeled glycoproteins.

    Science.gov (United States)

    Iskratsch, Thomas; Braun, Andreas; Paschinger, Katharina; Wilson, Iain B H

    2009-03-15

    Due to their ability to bind specifically to certain carbohydrate sequences, lectins are a frequently used tool in cytology, histology, and glycan analysis but also offer new options for drug targeting and drug delivery systems. For these and other potential applications, it is necessary to be certain as to the carbohydrate structures interacting with the lectin. Therefore, we used glycoproteins remodeled with glycosyltransferases and glycosidases for testing specificities of lectins from Aleuria aurantia (AAL), Erythrina cristagalli (ECL), Griffonia simplicifolia (GSL I-B(4)), Helix pomatia agglutinin (HPA), Lens culinaris (LCA), Lotus tetragonolobus (LTA), peanut (Arachis hypogaeae) (PNA), Ricinus communis (RCA I), Sambucus nigra (SNA), Vicia villosa (VVA), and wheat germ (Triticum vulgaris) (WGA) as well as reactivities of anti-carbohydrate antibodies (anti-bee venom, anti-horseradish peroxidase [anti-HRP], and anti-Lewis(x)). After enzymatic remodeling, the resulting neoglycoforms display defined carbohydrate sequences and can be used, when spotted on nitrocellulose or in enzyme-linked lectinosorbent assays, to identify the sugar moieties bound by the lectins. Transferrin with its two biantennary complex N-glycans was used as scaffold for gaining diverse N-glycosidic structures, whereas fetuin was modified using glycosidases to test the specificities of lectins toward both N- and O-glycans. In addition, alpha(1)-acid glycoprotein and Schistosoma mansoni egg extract were chosen as controls for lectin interactions with fucosylated glycans (Lewis(x) and core alpha1,3-fucose). Our data complement and expand the existing knowledge about the binding specificity of a range of commercially available lectins. PMID:19123999

  18. C-type natriuretic-derived peptides as biomarkers in human disease

    DEFF Research Database (Denmark)

    Lippert, Solvej Kølvraa; Goetze, Jens Peter

    2010-01-01

    and extracellular fluid volume. Atrial natriuretic peptide and B-type natriuretic peptide have gained considerable diagnostic interest as biomarkers in cardiovascular disease. By contrast, C-type natriuretic peptide has not yet been ascribed a role in human diagnostics. This perspective aims at......The natriuretic peptide system comprises three structurally related peptides: atrial natriuretic peptide, B-type natriuretic peptide and C-type natriuretic peptide. In circulation, they play an important endocrine role in the regulation of cardiovascular homeostasis by maintaining blood pressure...... recapitulating the present biochemical and clinical issues concerning C-type natriuretic peptide measurement in plasma as a potential biomarker....

  19. Tear fluid analysis in patients with primary Sjögren's syndrome using lectin probes

    DEFF Research Database (Denmark)

    Bjerrum, Kirsten Birgitte

    1999-01-01

    Ophthalmology, Sjögren's syndrome, dry eye, keratoconjunctivitis sicca, glycoprotein, mucus, lectins, Coomassie, electrophoresis, SDS-PAGE-blotting......Ophthalmology, Sjögren's syndrome, dry eye, keratoconjunctivitis sicca, glycoprotein, mucus, lectins, Coomassie, electrophoresis, SDS-PAGE-blotting...

  20. Mannose-binding lectin inhibits monocyte proliferation through transforming growth factor-β1 and p38 signaling pathways.

    Directory of Open Access Journals (Sweden)

    Yan Wang

    Full Text Available Mannose-binding lectin (MBL, a plasma C-type lectin, plays an important role in innate immunity. However, the interaction, and the consequences of it, between MBL and the immune system remain ill defined. We have investigated the contributing mechanisms and effects of MBL on the proliferation of human monocytes. At lower concentrations (≤4 μg/ml MBL was shown to partially enhance monocyte proliferation. By contrast, at higher concentrations (8-20 μg/ml of MBL, cell proliferation was markedly attenuated. MBL-induced growth inhibition was associated with G0/G1 arrest, down-regulation of cyclin D1/D3, cyclin-dependent kinase (Cdk 2/Cdk4 and up-regulation of the Cdk inhibitory protein Cip1/p21. Additionally, MBL induced apoptosis, and did so through caspase-3 activation and poly ADP-ribose polymerase (PARP cleavage. Moreover, transforming growth factor (TGF-β1 levels increased in the supernatants of MBL-stimulated monocyte cultures. We also found that MBL-dependent inhibition of monocyte proliferation could be reversed by the TGF-β receptor antagonist SB-431542, or by anti-TGF-β1 antibody, or by the mitogen-activated protein kinase (MAPK inhibitors specific for p38 (SB203580, but not ERK (U0126 or JNK (SP600125. Thus, at high concentrations, MBL can affect the immune system by inhibiting monocyte proliferation, which suggests that MBL may exhibit anti-inflammatory effects.

  1. Merging carbohydrate chemistry with lectin histochemistry to study inhibition of lectin binding by glycoclusters in the natural tissue context.

    Science.gov (United States)

    André, Sabine; Kaltner, Herbert; Kayser, Klaus; Murphy, Paul V; Gabius, Hans-Joachim

    2016-02-01

    Recognition of glycans by lectins leads to cell adhesion and growth regulation. The specificity and selectivity of this process are determined by carbohydrate structure (sequence and shape) and topology of its presentation. The synthesis of (neo)glycoconjugates with bi- to oligo-valency (glycoclusters) affords tools to delineate structure-activity relationships by blocking lectin binding to an artificial matrix, often a glycoprotein, or cultured cell lines. The drawback of these assays is that glycan presentation is different from that in tissues. In order to approach the natural context, we here introduce lectin histochemistry on fixed tissue sections to determine the susceptibility of binding of two plant lectins, i.e., GSA-II and WGA, to a series of 10 glycoclusters. Besides valency, this panel covers changes in the anomeric position (α/β) and the atom at the glycosidic linkage (O/S). Flanked by cell and solid-phase assays with human tumor lines and two mucins, respectively, staining (intensity and profile) was analyzed in sections of murine jejunum, stomach and epididymis as a function of glycocluster presence. The marked and differential sensitivity of signal generation to structural aspects of the glycoclusters proves the applicability of this method. This enables comparisons between data sets obtained by using (neo)glycoconjugates, cells and the tissue context as platforms. The special advantage of processing tissue sections is the monitoring of interference with lectin association at sites that are relevant for functionality. Testing glycoclusters in lectin histochemistry will especially be attractive in cases of multi-target recognition (glycans, proteins and lipids) by a tissue lectin. PMID:26553286

  2. Probing the cons and pros of lectin-induced immunomodulation: case studies for the mistletoe lectin and galectin-1.

    Science.gov (United States)

    Gabius, H J

    2001-07-01

    When imagining to monitor animal cells through a microscope with resolution at the molecular level, a salient attribute of their surfaces will be the abundance of glycan chains. They present galactosides at their termini widely extending like tentacles into the extracellular space. Their spatial accessibility and their potential for structural variability endow especially these glycan parts with capacity to act as docking points for molecular sensors (sugar receptors such as lectins). Binding and ligand clustering account for transmission of post-binding signals into the cell interior. The range of triggered activities has turned plant lectins into popular tools in cell biology and immunology. Potential for clinical application has been investigated rigorously only in recent years. As documented in vitro and in vivo for the galactoside-specific mistletoe lectin, its apparent immunomodulatory capacity reflected in upregulation of production of proinflammatory cytokines will not necessarily be clinically favorable but a double-edged sword. In fact, lectin application has been shown to stimulate tumor growth in cell lines, histocultures of human tumors and in two animal models using chemical carcinogenesis or tumor transplantation. When testing immunological effects of the endogenous lectin galectin-1, protection against disorders mediated by activated T cells came up for consideration. Elimination of these cells via CD7-dependent induction of apoptosis, and a shift to the Th2 response by the galectin, are factors to ameliorate disease states. This result encourages further efforts with other galectins. Functional redundancy, synergism, diversity or antagonism among galectins are being explored to understand the actual role of this class of endogenous lectins in inflammation. Regardless of the results of further preclinical testing for galectin-1, these two case studies break new ground in our understanding how glycans as ligands for lectins convey reactivity to

  3. C-type RNA viruses as possible predictors of radiation-induced bone oncogenesis

    International Nuclear Information System (INIS)

    Radium-224-induced osteosarcomas of (C3H X 101)F1 mice have been investigated for the presence of C-type RNA viruses. C-type virus particles with the typical properties of RNA tumour viruses could be detected by electron microscopic, biochemical and in-vitro studies. In a long-term experiment, the expression of C-type RNA virus particles was studied during the latency period and correlated with the appearance of osteosarcomas. The tumour incidence in this experiment was 100% with 40-50% of the tumours appearing in the 11th to 12th month after the start of the radiation. C-type RNA virus expression could be found in the 1st month and in the 11th and 12th months. These results indicate a transient virus activation shortly after starting the radiation, and a second peak of virus expression coinciding with the appearance of the osteosarcomas. (author)

  4. Biochimical and Structural caracterization of a lectin from Platypodium elegans Vogel seeds

    OpenAIRE

    Leite, Raquel,

    2011-01-01

    Lectin activity with specificity for mannose and glucose has been detected in the seed of Platypodium elegans, a legume from the Dalbergiae tribe. The gene of the lectin PELa has been cloned and the resulting 261 amino acid protein belongs to the legume lectin family with similarity with Pterocarpus angolensis agglutinin (PAL) from the same tribe. The recombinant lectin has been expressed in E. coli and refolded from inclusion bodies. Analysis of specificity by Glycan Array evidenced a very u...

  5. Toxicity and Binding Profile of Lectins from the Genus Canavalia on Brine Shrimp

    OpenAIRE

    Francisco Vassiliepe Sousa Arruda; Arthur Alves Melo; Mayron Alves de Vasconcelos; Romulo Farias Carneiro; Ito Liberato Barroso-Neto; Suzete Roberta Silva; Francisco Nascimento Pereira-Junior; Celso Shiniti Nagano; Kyria Santiago Nascimento; Edson Holanda Teixeira; Silvana Saker-Sampaio; Benildo Sousa Cavada; Alexandre Holanda Sampaio

    2013-01-01

    Lectins are sugar-binding proteins widely distributed in nature with many biological functions. Although many lectins have a remarkable biotechnological potential, some of them can be cytotoxic. Thus, the aim of this study was to assess the toxicity of five lectins, purified from seeds of different species of Canavalia genus. In order to determine the toxicity, assays with Artemia nauplii were performed. In addition, a fluorescence assay was carried out to evaluate the binding of lectins to A...

  6. Lectin histochemistry of palatine glands in the developing rat.

    Science.gov (United States)

    Hakami, Zaki; Kitaura, Hideki; Honma, Shiho; Wakisaka, Satoshi; Takano-Yamamoto, Teruko

    2014-05-01

    This study examined the binding pattern of lectins, soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin-I (UEA-I), peanut agglutinin (PNA), wheat germ agglutinin (WGA), and succinylated WGA (sucWGA) in the developing rat palatine glands. In adult rats, heterogeneous lectin binding patterns were revealed between the anterior and posterior portions of palatine glands, as DBA, VVA, and WGA were bound more intensely and broadly in the posterior portion. SBA, PNA, and sucWGA showed far less reactivity in the anterior than in the posterior portion. At embryonic day 18 (E18), weak labeling was observed with UEA-I and WGA at the basal membrane of terminal buds, UEA-I and PNA labeled the epithelial cord, and there was no apparent binding for SBA, DBA, VVA, and sucWGA. At E20, after acinar lumenization, all lectins were detected at the acinar cell basal membranes. After birth, all lectins detectably labeled at the mucous cell apical membranes and progressively, with maturation, extended from the apical to basal portions of the cytoplasm. Apparent serous cells were observed around postnatal day 10 (PN10) and bound UEA-I. Lectins reached peak reactivity at PN21 and the binding patterns became identical to those of adults around PN28. PMID:24345684

  7. C-type natriuretic peptide modulates permeability of the blood–brain barrier

    OpenAIRE

    BOHARA, Manoj; Kambe, Yuki; Nagayama, Tetsuya; TOKIMURA, Hiroshi; Arita, Kazunori; Miyata, Atsuro

    2014-01-01

    C-type natriuretic peptide (CNP) is abundant in brain and is reported to exert autocrine function in vascular cells, but its effect on blood–brain barrier (BBB) permeability has not been clarified yet. Here, we examined this effect. Transendothelial electrical resistance (TEER) of in vitro BBB model, composed of bovine brain microvascular endothelial cells and astrocytes, was significantly dose dependently decreased by CNP (1, 10, and 100 nmol/L). C-type natriuretic peptide treatment reduced ...

  8. 透明质酸对关节软骨蛋白聚糖aggrecan的影响%Effect of hyaluronic acid on articular cartilage proteoglycan aggrecan

    Institute of Scientific and Technical Information of China (English)

    杜国辉; 屈爱存; 陈建英; 贺艳丽

    2011-01-01

    目的 研究关节腔注射透明质酸对兔骨关节炎软骨基质的保护作用.方法 关节骨液用软骨素酶ABC降解,丹磺酰肼柱前衍生.采用NH2色谱柱,流动相为乙腈-10 mmol/L醋酸溶液-10 mmol/L醋酸钠溶液(7616:8),荧光检测器检测,激发波长350 nm,发射波长530 nm,流速为1mL/min.结果 透明质酸可减少兔关节滑液中C4S、C6S的含量.结论 关节腔注射透明质酸可抑制兔软骨蛋白聚糖的降解或上调蛋白聚糖的合成,维持关节软骨正常的新陈代谢.%Purpose To investigate the protective effect of intra-articular injection of hyaluronic acid on cartilage matrix in rabbit osteoarthritis. Methods Joint synovial fluid was degraded by chondroitinase ABC, and then derived with dansylhydrazine. The derivatized sample was separated on NH2 column. The mobile phase of acetonitrile- acetic acid- sodium acetate (76:16:8) was used. Detected with fluorescence detector, the excitation wavelength (λex) and emission wavelength (λem) were 350 and 530 nm, respectively. The flow rate was 1.0 mL/min. Results Hyaluronic acid could decrease the contents of C4S and C6S in rabbit joint synovial fluid. Conclusion The intra-articular injection of hyaluronic acid can reduce the degradation of aggrecan and/or enhance the synthesis of aggrecan in the joint tissues of rabbit osteoarthritis, and maintain the normal metabolism of articular cartilage.

  9. Histological and lectin histochemical studies on the olfactory and respiratory mucosae of the sheep.

    Science.gov (United States)

    Ibrahim, Dalia; Nakamuta, Nobuaki; Taniguchi, Kazumi; Yamamoto, Yoshio; Taniguchi, Kazuyuki

    2014-03-01

    The olfactory and respiratory mucosae of the Corriedale sheep were examined using lectin histochemistry in order to clarify the histochemical and glycohistochemical differences between these two tissues. The olfactory epithelium was stained with 13 lectins out of 21 lectins examined, while the respiratory epithelium was positive to 16 lectins. The free border of both of the olfactory and respiratory epithelia was stained with 12 lectins: Wheat germ agglutinin (WGA), succinylated-wheat germ agglutinin (s-WGA), Lycopersicon esculentum lectin (LEL), Solanum tuberosum lectin (STL), Datura stramonium lectin (DSL), Soybean agglutinin (SBA), Bandeiraea simplicifolia lectin-I (BSL-I), Ricinus communis agglutinin-I (RCA-120), Erythrina cristagalli lectin (ECL), Concanavalin A (Con A), Phaseolus vulgaris agglutinin-E (PHA-E) and Phaseolus vulgaris agglutinin-L (PHA-L). The associated glands of the olfactory mucosa, Bowman's glands, were stained with 13 lectins. While both the goblet cells and mucous nasal glands were stained with 8 lectins; five of them (WGA, s-WGA, STL, Vicia villosa agglutinin (VVA) and ECL) were mutually positive among the Bowman's glands, mucous nasal glands and the goblet cells. These findings indicate that the glycohistochemical characteristics of the free borders of both olfactory and respiratory epithelia are similar to each other, suggesting that secretions from the Bowman's glands and those of the goblet cells and mucous nasal glands are partially exchanged between the surface of two epithelia to contribute the functions of the respiratory epithelium and the olfactory receptor cells, respectively. PMID:24200894

  10. The ovine newborn and human foetal intervertebral disc contain perlecan and aggrecan variably substituted with native 7D4 CS sulphation motif: spatiotemporal immunolocalisation and co-distribution with Notch-1 in the human foetal disc.

    Science.gov (United States)

    Shu, Cindy; Hughes, Clare; Smith, Susan M; Smith, Margaret M; Hayes, Anthony; Caterson, Bruce; Little, Christopher B; Melrose, James

    2013-10-01

    Composite agarose (1.2 %) polyacrylamide (0.6 %) gel electrophoresis was used to separate discrete populations of native aggrecan and perlecan in newborn to 10 year old ovine intervertebral discs (IVDs). Semi-dry immunoblotting using core-protein and glycosaminoglycan (GAG) side chain specific monoclonal antibodies in combination with chondroitin ABC lyase demonstrated intra-chain native 7-D-4 chondroitin sulphate (CS) sulphation motifs and variable proportions of non-reducing terminal Δ4,5-unsaturated uronate-N-acetylgalactosamine-4-sulphate [2B6(+)] and Δ4,5-unsaturated glucuronate-N-acetylgalactosamine-6-sulphate [3B3(+)] disaccharides. The relative abundance of 2-B-6(+) aggrecan increased with advancing age of the IVD samples while the converse was true for the 3-B-3(+) aggrecan population. Relative 7D4 levels in aggrecan and perlecan were highest in the newborn IVD and significantly lower in the older IVD and other cartilage samples. Quantitation of 7D4 proteoglycan by enzyme linked immunosorbent inhibition assay confirmed the newborn ovine nucleus pulposus (NP) and inner annulus fibrosus (AF) contained higher levels (1.2-1.32 μg 7-D-4-proteoglycan/mg tissue wet weight) than the 2 (0.35-0.42 μg/mg wet weight tissue) and 10 year old IVD samples (0.16-0.22 μg/mg tissue wet weight) with the outer AF zones consistently containing lower levels of 7-D-4 epitope in all cases (P < 0.001). Cell populations on the margins of the AF and cartilaginous vertebral rudiments in newborn ovine and human foetal IVD strongly expressed 7-D-4 CS epitope and perlecan, This was co-distributed with Notch-1 expression in human foetal IVDs consistent with the 7-D-4 CS sulphation motif representing a marker of tissue development expressed by disc progenitor cell populations. PMID:23756834

  11. A novel C-type lysozyme from Mytilus galloprovincialis: insight into innate immunity and molecular evolution of invertebrate C-type lysozymes.

    Directory of Open Access Journals (Sweden)

    Qing Wang

    Full Text Available A c-type lysozyme (named as MgCLYZ gene was cloned from the mussel Mytilus galloprovincialis. Blast analysis indicated that MgCLYZ was a salivary c-type lysozyme which was mainly found in insects. The nucleotide sequence of MgCLYZ was predicted to encode a polypeptide of 154 amino acid residues with the signal peptide comprising the first 24 residues. The deduced mature peptide of MgCLYZ was of a calculated molecular weight of 14.4 kD and a theoretical isoelectric point (pI of 8.08. Evolution analysis suggested that bivalve branch of the invertebrate c-type lysozymes phylogeny tree underwent positive selection during evolution. By quantitative real-time RT-PCR (qRT-PCR analysis, MgCLYZ transcript was widely detected in all examined tissues and responded sensitively to bacterial challenge in hemocytes and hepatopancreas. The optimal temperature and pH of recombinant MgCLYZ (rMgCLYZ were 20°C and 4, respectively. The rMgCLYZ displayed lytic activities against Gram-positive bacteria including Micrococcus luteus and Staphyloccocus aureus, and Gram-negative bacteria including Vibrio anguillarum, Enterobacter cloacae, Pseudomonas putida, Proteus mirabilis and Bacillus aquimaris. These results suggest that MgCLYZ perhaps play an important role in innate immunity of M. galloprovincialis, and invertebrate c-type lysozymes might be under positive selection in a species-specific manner during evolution for undergoing adaptation to different environment and diverse pathogens.

  12. Isolation and Biochemical Characterization of Apios Tuber Lectin

    Directory of Open Access Journals (Sweden)

    Eri Kenmochi

    2015-01-01

    Full Text Available Apios tuber lectin, named ATL, was isolated from Apios americana Medikus by two chromatography steps, hydrophobic chromatography and anion-exchange chromatography. The minimum concentration required for the hemagglutination activity toward rabbit erythrocytes of ATL was 4 μg/mL. ATL was composed of a homodimer of 28.4 kDa subunits. The amino acid sequence of ATL was similar to those of other legume lectins. The lectin showed moderate stability toward heating and acidic pH, and the binding affinity against several monosaccharides, such as D-glucosamine and D-galactosamine. ATL also bound to desialylated or agalactosylated glycoproteins such as asialo and agalacto transferrin. ATL decreased the transepithelial electrical resistance across human intestinal Caco-2 cell monolayers, suggesting the effect on the tight junction-mediated paracellular transport.

  13. Purification of a thermostable antinociceptive lectin isolated from Andira anthelmia.

    Science.gov (United States)

    Nascimento, Kyria Santiago; Nascimento, Francisco Lucas Faustino do; Silva, Mayara Torquato Lima; Nobre, Camila Bezerra; Moreira, Cleane Gomes; Brizeno, Luiz André Cavalcante; da Ponte, Edson Lopes; Assreuy, Ana Maria Sampaio; Cavada, Benildo Sousa

    2016-06-01

    Andira anthelmia (tribe Dalbergieae), a plant from Brazilian Amazon, possesses a seed lectin that was purified by affinity chromatography in sepharose-mannose. This novel Dalbergieae lectin, named AAL, agglutinated rabbit erythrocytes treated with trypsin. The hemagglutinating activity of AAL was maintained after incubation at a wide range of temperature (40 to 70 °C) and pH, was shown to be dependent on divalent cations, and was inhibited by d-mannose and d-sucrose. AAL showed an electrophoretic profile in sodium dodecyl sulfate-polyacrylamide gel electrophoresis similar to other lectins of the tribe Dalbergieae, presenting a double band of molecular weight with approximately 20 kDa and other minor bands of 17, 15, and 13 kDa, being the smaller fragment glycosylated. AAL injected by intravenous route in mice showed antinociceptive activity in two behavioral tests (writhing and formalin). In the writhing test induced by acetic acid, AAL showed inhibitory effect at 0.01 mg/kg (68%), 0.1 mg/kg (46%) and 1 mg/kg (74%). In the formalin test, AAL (0.1 mg/kg) inhibited by 48% the licking time in the inflammatory phase, an effect that was recovered by the lectin association with mannose. In conclusion, AAL presents analgesic effect involving the lectin domain via peripheral mechanisms of inflammatory nociception. This activity highlights the importance of lectins as tools to be used for understanding the interaction of protein-carbohydrate in processes associated to inflammatory pain. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26638121

  14. Effect of Lectins from Diocleinae Subtribe against Oral Streptococci

    Directory of Open Access Journals (Sweden)

    Edson Holanda Teixeira

    2011-04-01

    Full Text Available Surface colonization is an essential step in biofilm development. The ability of oral pathogens to adhere to tooth surfaces is directly linked with the presence of specific molecules at the bacterial surface that can interact with enamel acquired pellicle ligands. In light of this, the aim of this study was to verify inhibitory and antibiofilm action of lectins from the Diocleinaesubtribe against Streptococcus mutans and Streptococcus oralis. The inhibitory action against planctonic cells was assessed using lectins from Canavaliaensi formis (ConA, Canavalia brasiliensis (ConBr, Canavalia maritima (ConM, Canavalia gladiata (CGL and Canavalia boliviana (ConBol. ConBol, ConBr and ConM showed inhibitory activity on S. mutans growth. All lectins, except ConA, stimulated significantly the growth of S. oralis. To evaluate the effect on biofilm formation, clarified saliva was added to 96-well, flat-bottomed polystyrene plates, followed by the addition of solutions containing 100 or 200 µg/mL of the selected lectins. ConBol, ConM and ConA inhibited the S. mutans biofilms. No effects were found on S. oralis biofilms. Structure/function analysis were carried out using bioinformatics tools. The aperture and deepness of the CRD (Carbohydrate Recognition Domain permit us to distinguish the two groups of Canavalia lectins in accordance to their actions against S. mutans and S. oralis. The results found provide a basis for encouraging the use of plant lectins as biotechnological tools in ecological control and prevention of caries disease.

  15. Ferromagnetic Levan Composite: An Affinity Matrix to Purify Lectin

    Directory of Open Access Journals (Sweden)

    Renata Angeli

    2009-01-01

    Full Text Available A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A and Cratylia mollis (Cramoll 1 and Cramoll 1,4 did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column.

  16. Ferromagnetic levan composite: an affinity matrix to purify lectin.

    Science.gov (United States)

    Angeli, Renata; da Paz, Nathalia V N; Maciel, Jackeline C; Araújo, Flávia F B; Paiva, Patrícia M G; Calazans, Glícia M T; Valente, Ana Paula; Almeida, Fábio C L; Coelho, Luana C B B; Carvalho, Luiz B; Silva, Maria da Paz C; dos Santos Correia, Maria Tereza

    2009-01-01

    A simple and inexpensive procedure used magnetite and levan to synthesize a composite recovered by a magnetic field. Lectins from Canavalia ensiformis (Con A) and Cratylia mollis (Cramoll 1 and Cramoll 1, 4) did bind specifically to composite. The magnetic property of derivative favored washing out contaminating proteins and recovery of pure lectins with glucose elution. Cramoll 1 was purified by this affinity binding procedure in two steps instead of a previous three-step protocol with ammonium sulfate fractionation, affinity chromatography on Sephadex G-75, and ion exchange chromatography through a CM-cellulose column. PMID:19547713

  17. INTERACTION BETWEEN THE SURFACE GLYCOSYLATED POLYPROPYLENE MEMBRANE AND LECTIN

    Institute of Scientific and Technical Information of China (English)

    Qian Yang; Ling-shu Wan; Zhi-kang Xu

    2008-01-01

    A glycopolymer bearing glucose residues was tethered onto the surface of polypropylene microporous membrane by UV-induced graft polymerization of α-allyl glucoside. Concanavalin A (Con A), a glucose recognizing lectin, could be specifically adsorbed to the membrane surface. On the other hand, the membrane surface showed no recognition ability to another lectin peanut agglutinin. Moreover, the recognition complex between the glycosylated membrane surface and Con Acould be inhibited by glucose and mannose solution. This surface glycosylated membrane could be used as affinity membrane for protein separation and purification.

  18. The galactophilic lectin (PA-IL, gene LecA) from Pseudomonas aeruginosa. Its binding requirements and the localization of lectin receptors in various mouse tissues

    DEFF Research Database (Denmark)

    Kirkeby, Svend; Hansen, Axel K; d'Apice, Anthony;

    2006-01-01

    The opportunistic pathogen Pseudomonas aeruginosa contains lectins of which one of them, PA-IL (gene lecA), shows preference for alpha-galactosylated glycans. The bacterial lectin is probably important in the carbohydrate-mediated adhesion of the microorganism to endothelia and epithelia and...... thereby the lectin facilitates entering and damaging of the cells. The requirements for the interaction between PA-IL and the carbohydrate epitopes to which the bacterial lectin may bind were here Studied using alpha-galactosylated neoglycoproteins that were immobilized on Microtiter plates. It is...... concluded that the carbohydrate recognizing site of the lectin can have a binding requirement of only one saccharide. Lectin histochemistry was performed on sections from wild type mice and from knock-out mice, which lack function of the alpha.1,3-galactosyltransferase gene. All assays with the P...

  19. Interplay between metal binding and cis/trans isomerization in legume lectins: structural and thermodynamic study of P. angolensis lectin.

    Science.gov (United States)

    Garcia-Pino, Abel; Buts, Lieven; Wyns, Lode; Loris, Remy

    2006-08-01

    The interplay between metal binding, carbohydrate binding activity, stability and structure of the lectin from Pterocarpus angolensis was investigated. Removal of the metals leads to a more flexible form of the protein with significantly less conformational stability. Crystal structures of this metal-free form show significant structural rearrangements, although some structural features that allow the binding of sugars are retained. We propose that substitution of an asparagine residue at the start of the C-terminal beta-strand of the legume lectin monomer hinders the trans-isomerization of the cis-peptide bond upon demetallization and constitutes an intramolecular switch governing the isomer state of the non-proline bond and ultimately the lectin phenotype. PMID:16824540

  20. 树突状细胞相关凝集素受体1和2在真菌免疫领域的研究进展%DC-associated C-type lecxin-1 and DC-associated C-type lecxin-2 in antifungal dfences

    Institute of Scientific and Technical Information of China (English)

    黄晓强; 陈剑

    2012-01-01

    树突状细胞相关凝集素受体1和2(Dectin1和2)是2C型凝集素受体家族(CLR)的重要成员,作为模式识别受体( PRRs),其有效地识别病原体相关分子模式(PAMPs);Dectin-1识别β-葡聚糖,通过自身免疫受体络氨酸激活基序( ITAM)向胞内转导信号;Dectin-2识别α-甘露聚糖,通过耦联FcRγ链上的ITAM结构转导信号.ITAM招募并激活非受体络氨酸蛋白激酶(Syk),后者激活MAPKs或介导CARD9-Malt1-Bcl10复合体组装,激活核因子-κB(NF-κB),诱导合成炎症因子等一系列细胞活动.其配体β-葡聚糖和甘露聚糖都是真菌细胞壁的主要成分.近年来研究表明,Dectin-1和Dectin-2受体在真菌免疫防御中具有重要作用.%DC-associated C-type lectin-1 and 2 (Dectin-1 and Dectin-2) are pivotal C-type lectin family members,both of them function as pattern recognition receptors (PRRs) to initiate innate and adaptive immunity by recognizing pathogen-associated molecular patterns (PAMPs).Dectin-1 recognizes β-glucans and signals with its cytoplasmic immunoreceptor tyrosine-based activation motif (ITAM)-like motif,whereas Dectin-2 recognizes α-mannans and signals through association with the ITAM-containing Fc receptor γ chain.Upon ligand ligation,spleen tyrosine kinase (Syk) is recruited to the ITAM.Syk mediates the activation of MAPKs;or induces the formation of the CARD9-Malt1-Bcl10 complex,resulting in the activation of nuclear factor-κB (NF-κB).Recert studies indicate that Dectin-1 and Dectin-2 play important roles in immune defense against fungi.

  1. Galactose/N-acetylgalactosamine lectin: the coordinator of host cell killing

    Indian Academy of Sciences (India)

    Douglas R Boettner; Christopher Huston; William A Petri Jr

    2002-11-01

    Entamoeba histolytica is an enteric parasite that can kill host cells via a contact-dependent mechanism. This killing involves the amoebic surface protein referred to as the Gal/GalNAc lectin. The Gal/GalNAc lectin binds galactose and N-acetylgalactosamine allowing the adherence of amoebas to host cells. Involvement of the lectin in the pathogenesis of E. histolytica infection will be reviewed in this paper. The lectin has been shown to have very specific and substantial effects on adherence, cytotoxicity, and encystation. There is also possible involvement of the lectin in phagocytosis and caspase activation in host cells.

  2. Comparison of docking methods for carbohydrate binding in calcium-dependent lectins and prediction of the carbohydrate binding mode to sea cucumber lectin CEL-III

    OpenAIRE

    Nurisso, Alessandra; Kozmon, Stanislav; Imberty, Anne

    2008-01-01

    Abstract Lectins display a variety of strategies for specific recognition of carbohydrates. In several lectin families from different origin, one or two calcium ions are involved in the carbohydrate binding site with direct coordination of the sugar hydroxyl groups. Our work implied a molecular docking study involving a set of bacterial and animal calcium-dependant lectins in order to compare the ability of three docking programs to reproduce key carbohydrate-metal interactions. Fl...

  3. Association Study of Mannose-Binding Lectin Levels and Genetic Variants in Lectin Pathway Proteins with Susceptibility to Age-Related Macular Degeneration: A Case-Control Study

    OpenAIRE

    Osthoff, Michael; Dean, Melinda M.; Baird, Paul N.; Richardson, Andrea J.; Daniell, Mark; Guymer, Robyn H.; Eisen, Damon P

    2015-01-01

    Background In age-related macular degeneration (AMD) the complement system is thought to be activated by chronic oxidative damage with genetic variants identified in the alternative pathway as susceptibility factors. However, the involvement of the lectin pathway of complement, a key mediator of oxidative damage, is controversial. This study investigated whether mannose-binding lectin (MBL) levels and genetic variants in lectin pathway proteins, are associated with the predisposition to and s...

  4. Reciprocal voltage sensor-to-pore coupling leads to potassium channel C-type inactivation

    Science.gov (United States)

    Conti, Luca; Renhorn, Jakob; Gabrielsson, Anders; Turesson, Fredrik; Liin, Sara I.; Lindahl, Erik; Elinder, Fredrik

    2016-06-01

    Voltage-gated potassium channels open at depolarized membrane voltages. A prolonged depolarization causes a rearrangement of the selectivity filter which terminates the conduction of ions – a process called slow or C-type inactivation. How structural rearrangements in the voltage-sensor domain (VSD) cause alteration in the selectivity filter, and vice versa, are not fully understood. We show that pulling the pore domain of the Shaker potassium channel towards the VSD by a Cd2+ bridge accelerates C-type inactivation. Molecular dynamics simulations show that such pulling widens the selectivity filter and disrupts the K+ coordination, a hallmark for C-type inactivation. An engineered Cd2+ bridge within the VSD also affect C-type inactivation. Conversely, a pore domain mutation affects VSD gating-charge movement. Finally, C-type inactivation is caused by the concerted action of distant amino acid residues in the pore domain. All together, these data suggest a reciprocal communication between the pore domain and the VSD in the extracellular portion of the channel.

  5. Reciprocal voltage sensor-to-pore coupling leads to potassium channel C-type inactivation

    Science.gov (United States)

    Conti, Luca; Renhorn, Jakob; Gabrielsson, Anders; Turesson, Fredrik; Liin, Sara I; Lindahl, Erik; Elinder, Fredrik

    2016-01-01

    Voltage-gated potassium channels open at depolarized membrane voltages. A prolonged depolarization causes a rearrangement of the selectivity filter which terminates the conduction of ions – a process called slow or C-type inactivation. How structural rearrangements in the voltage-sensor domain (VSD) cause alteration in the selectivity filter, and vice versa, are not fully understood. We show that pulling the pore domain of the Shaker potassium channel towards the VSD by a Cd2+ bridge accelerates C-type inactivation. Molecular dynamics simulations show that such pulling widens the selectivity filter and disrupts the K+ coordination, a hallmark for C-type inactivation. An engineered Cd2+ bridge within the VSD also affect C-type inactivation. Conversely, a pore domain mutation affects VSD gating-charge movement. Finally, C-type inactivation is caused by the concerted action of distant amino acid residues in the pore domain. All together, these data suggest a reciprocal communication between the pore domain and the VSD in the extracellular portion of the channel. PMID:27278891

  6. The use of lectin microarray for assessing glycosylation of therapeutic proteins.

    Science.gov (United States)

    Zhang, Lei; Luo, Shen; Zhang, Baolin

    2016-04-01

    Glycans or carbohydrates attached to therapeutic glycoproteins can directly affect product quality, safety and efficacy, and therefore must be adequately analyzed and controlled throughout product life cycles. However, the complexity of protein glycosylation poses a daunting analytical challenge. In this study, we evaluated the utility of a lectin microarray for assessing protein glycans. Using commercial lectin chips, which contain 45 lectins toward distinct glycan structures, we were able to determine the lectin binding patterns of a panel of 15 therapeutic proteins, including 8 monoclonal antibodies. Lectin binding signals were analyzed to generate glycan profiles that were generally consistent with the known glycan patterns for these glycoproteins. In particular, the lectin-based microarray was found to be highly sensitive to variations in the terminal carbohydrate structures such as galactose versus sialic acid epitopes. These data suggest that lectin microarray could be used for screening glycan patterns of therapeutic glycoproteins. PMID:26918373

  7. Regulation of Adherence and Virulence by the Entamoeba histolytica Lectin Cytoplasmic Domain, Which Contains a β2 Integrin Motif

    OpenAIRE

    Vines, Richard R.; Ramakrishnan, Girija; Rogers, Joshua B.; Lockhart, Lauren A.; Mann, Barbara J.; Petri, William A.

    1998-01-01

    Killing of human cells by the parasite Entamoeba histolytica requires adherence via an amebic cell surface lectin. Lectin activity in the parasite is regulated by inside-out signaling. The lectin cytoplasmic domain has sequence identity with a region of the β2 integrin cytoplasmic tail implicated in regulation of integrin-mediated adhesion. Intracellular expression of a fusion protein containing the cytoplasmic domain of the lectin has a dominant negative effect on extracellular lectin-mediat...

  8. Mannan-binding lectin in astma and allergy

    DEFF Research Database (Denmark)

    Kaur, S.; Thiel, Steffen; Sarma, P.U.;

    2006-01-01

    Mannan-binding lectin (MBL) is a vital and versatile component of innate immunity. It is present in serum and may bind to a plethora of microbial pathogens and mediate opsonization of these by complement-dependent and/or independent mechanisms. Low-MBL levels in serum, attributed to certain genetic...

  9. Clinical manifestations of mannan-binding lectin deficiency

    DEFF Research Database (Denmark)

    Thiel, Steffen; Frederiksen, Pernille Dorthea; Jensenius, Jens Christian

    2006-01-01

    Mannan-binding lectin (MBL) is a plasma protein of the innate immune system with the ability to initiate antimicrobial and inflammatory actions. MBL deficiency is common. More than 10% of the general population may, depending on definition, be classified as MBL deficient, underlining the redundancy...

  10. Cancer Biomarker Discovery: Lectin-Based Strategies Targeting Glycoproteins

    Directory of Open Access Journals (Sweden)

    David Clark

    2012-01-01

    Full Text Available Biomarker discovery can identify molecular markers in various cancers that can be used for detection, screening, diagnosis, and monitoring of disease progression. Lectin-affinity is a technique that can be used for the enrichment of glycoproteins from a complex sample, facilitating the discovery of novel cancer biomarkers associated with a disease state.

  11. Architectures of Multivalent Glycomimetics for Probing Carbohydrate-Lectin Interactions

    Science.gov (United States)

    Lahmann, Martina

    Well-defined multivalent glycoconjugates are valued tools in glycoscience and they are particularly valuable for the investigation of carbohydrate-lectin interactions. In addition to the relatively globularly shaped glycodendrimers many other designs have been realized. This chapter gives an overview on the common different architectures and their chemical synthesis by focussing on the achievements made since 2001.

  12. Mannose-Binding Lectin Deficiency Is Associated with Myocardial Infarction

    DEFF Research Database (Denmark)

    Vengen, Inga Thorsen; Madsen, Hans O; Garred, Peter;

    2012-01-01

    Mannose-binding lectin (MBL) and ficolins activate the complement cascade, which is involved in atherogenesis. Based on a pilot study, we hypothesized that functional polymorphisms in the MBL gene (MBL2) leading to dysfunctional protein are related to development of myocardial infarction (MI). The...

  13. Momordica charantia seed lectin: toxicity, bacterial agglutination and antitumor properties.

    Science.gov (United States)

    Kabir, Syed Rashel; Nabi, Md Mahamodun; Nurujjaman, Md; Abu Reza, Md; Alam, A H M Khurshid; Uz Zaman, Rokon; Khalid-Bin-Ferdaus, Khandaker Md; Amin, Ruhul; Khan, Md Masudul Hasan; Hossain, Md Anowar; Uddin, Md Salim; Mahmud, Zahid Hayat

    2015-03-01

    In last three decades, several studies were carried out on the D-galactose-specific lectin of Momordica charantia seeds (MCL). In the present study, in vitro growth inhibition (8-23 %) at different concentrations (6-24 μg/ml) of MCL was observed against Ehrlich ascites carcinoma (EAC) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. MCL also showed 28, 45, and 75 % growth inhibitions against EAC cells when administered 1.2, 2.0, and 2.8 mg/kg/day (i.p.), respectively for five consequent days in vivo in mice. After lectin treatment, the level of red blood cell and hemoglobin was increased significantly with the decrease of white blood cell and maintained the normal level when compared with EAC-bearing control and normal mice without EAC cells. Although MCL caused cell cycle arrest at G0/G1 phase of EAC cells, any irregular shape or apoptotic morphological alterations in the lectin-treated EAC cells was not observed by an optical and fluorescence microscope. Lectin showed toxicity against brine shrimp nauplii with an LC50 value of 49.7 μg/ml. Four out of seven pathogenic bacteria were agglutinated by MCL in the absence of inhibitory sugar D-lactose/D-galactose. In conclusion, MCL showed strong cytotoxic effect and therefore can be used as a potent anticancer chemotherapeutic agent. PMID:25542240

  14. Preparation and identification of 1.3 copies C-type HBV transgenic mice

    OpenAIRE

    Chen, Mei-Juan; Yu-qin YOU; Guang-ze LIU; Tong, Ming-Hua; Jing-wei LI; Xiu-mei LI; Kong, Xiang-Ping

    2011-01-01

    Objective To prepare 1.3 copies C-type HBV transgenic mice for providing a better model for the prevention and treatment of hepatitis B.Methods The HBV transgenic mice were generated by microinjection of 1.3 copies C-type HBV genome into the pronucleus of FVB /N zygotes.PCR,ELISA,RT-PCR and immunohistochemistry were used to detect the integration,replication and expression of HBV gene in the transgenic mice.Results Tow thousand two hundred and eighty-two fertilized eggs were injected and a to...

  15. Mannan-binding lectin and mannan-binding lectin-associated serine protease 2 in acute pancreatitis

    DEFF Research Database (Denmark)

    Novovic, Srdan; Andersen, Anders; Ersbøll, Annette Kjær;

    2011-01-01

    Complement activation may play a prominent role in acute pancreatitis (AP). Mannan-binding lectin (MBL) and MBL-associated serine protease 2 (MASP-2) participate in complement activation. The objective of the present study was to evaluate the role of MBL and MASP-2 as markers in AP with regard to...

  16. Sialic acid specific lectins from Episesarma tetragonum (Decapoda, Grapsidae: isolation, purification and characterization

    Directory of Open Access Journals (Sweden)

    R. Viswambari Devi

    2013-07-01

    Full Text Available Two sialic acid specific lectins Episesarma tetragonum agglutinin–1 and 2 were purified from the hemolymph of the Mangrove crab, Episesarma tetragonum. The major lectin was purified using CNBr-activated sepharose 4B conjugated to fetuin. N-acetyl glucosamine containing buffer was used for elution. The hemagglutination activity of purified lectin was inhibited by glycoproteins containing Siaα, 2-3Galβ, 1-4 GlcNAc linkages. On SDS-PAGE, the molecular weight of calcium dependent lectin was observed to be 70 kDa. Lectin had the maximum activity at a wide range of pH (6.5 – 9.5 and temperature (0 - 40 °C.  The physicochemical characteristics of the minor agglutinin showed that its hemagglutinating activity was calcium dependent, optimum at pH 8 – 9.5 and temperature 0 – 37 °C. The only potent inhibitor of minor lectin was bovine submaxillary mucin. An attempt was also made to purify minor lectin by affinity chromatography using bovine submaxillary mucin coupled to CNBr-activated sepharose 4B column. The lectin was eluted with elution buffer containing ethylene diamine tetra acetate. Strong inhibition of purified minor lectin by bovine submaxillary mucin and non-inhibitory action of de-O-acetylated bovine submaxillary mucin suggested that the lectin was O-acetyl sialic acid specific.

  17. Functional environmental proteomics: elucidating the role of a c-type cytochrome abundant during uranium bioremediation.

    Science.gov (United States)

    Yun, Jiae; Malvankar, Nikhil S; Ueki, Toshiyuki; Lovley, Derek R

    2016-02-01

    Studies with pure cultures of dissimilatory metal-reducing microorganisms have demonstrated that outer-surface c-type cytochromes are important electron transfer agents for the reduction of metals, but previous environmental proteomic studies have typically not recovered cytochrome sequences from subsurface environments in which metal reduction is important. Gel-separation, heme-staining and mass spectrometry of proteins in groundwater from in situ uranium bioremediation experiments identified a putative c-type cytochrome, designated Geobacter subsurface c-type cytochrome A (GscA), encoded within the genome of strain M18, a Geobacter isolate previously recovered from the site. Homologs of GscA were identified in the genomes of other Geobacter isolates in the phylogenetic cluster known as subsurface clade 1, which predominates in a diversity of Fe(III)-reducing subsurface environments. Most of the gscA sequences recovered from groundwater genomic DNA clustered in a tight phylogenetic group closely related to strain M18. GscA was most abundant in groundwater samples in which Geobacter sp. predominated. Expression of gscA in a strain of Geobacter sulfurreducens that lacked the gene for the c-type cytochrome OmcS, thought to facilitate electron transfer from conductive pili to Fe(III) oxide, restored the capacity for Fe(III) oxide reduction. Atomic force microscopy provided evidence that GscA was associated with the pili. These results demonstrate that a c-type cytochrome with an apparent function similar to that of OmcS is abundant when Geobacter sp. are abundant in the subsurface, providing insight into the mechanisms for the growth of subsurface Geobacter sp. on Fe(III) oxide and suggesting an approach for functional analysis of other Geobacter proteins found in the subsurface. PMID:26140532

  18. Mannan-binding lectin and healing of a radiation-induced chronic ulcer--a case report on mannan-binding lectin replacement therapy

    DEFF Research Database (Denmark)

    Maaløe, Nanna; Bonde, C; Laursen, I;

    2011-01-01

    Mannan-binding lectin is an important component of innate immunity, and insufficiency is associated with several clinical disorders. Recently, experimental replacement therapy with plasma-derived mannan-binding lectin has become an option. The current article presents the case of a patient with an...

  19. Purification, partial characterization and antimicrobial activity of Lectin from Chenopodium Quinoa seeds

    Directory of Open Access Journals (Sweden)

    Dávia Guimarães POMPEU

    2015-01-01

    Full Text Available Abstract A novel lectin was isolated from the seeds of Chenopodium quinoa. To achieve this end, the crude extract from the quinoa was submitted to two purification steps, Sephadex G50 and Mono Q. The hemagglutinating activity showed that this lectin agglutinates human erythrocytes. Its activity is inhibited by glucose and mannose, and remained stable under a wide range of pH levels and temperatures. The quinoa lectin was found to be a heterodimeric lectin of approximately 60 kDa, consisting of two subunits of approximately 25 kDa and 35 kDa. This lectin had its antimicrobial activity tested against several bacteria strains and effectively inhibited three strains. These strains were all Gram-negative, making this lectin a promising antimicrobial tool.

  20. Differential lectin binding to cellular membranes in the epidermis of the newborn rat.

    OpenAIRE

    Brabec, R K; Peters, B P; Bernstein, I A; Gray, R H; Goldstein, I. J.

    1980-01-01

    Three fluorescein-labeled lectins have been shown to exhibit specificity for the surface of cells in different layers of the epidermis in the newborn rat. An isolectin from seeds of Bandeiraea simplicifolia with specificity for alpha-D-galactosyl end groups labeled the basal and lower spinous cells; a lectin from Ulex europaeus exhibiting specificity for alpha-L-fucosyl units outlines the surface of spinous cells, and a second lectin from B. simplicifolia, with specificity for N-acetyl-D-gluc...

  1. Stability, subunit interactions and carbohydrate-binding of the seed lectin from Pterocarpus angolensis.

    Science.gov (United States)

    Echemendia-Blanco, Dannele; Van Driessche, Edilbert; Ncube, Ignatious; Read, John S; Beeckmans, Sonia

    2009-01-01

    From 1 kg of defatted Pterocarpus angolensis (mukwa tree) seed meal, 21.6 grams of an alpha,D-mannose/glucose-specific lectin can be purified on mannose-Sepharose. Relative affinities for several (oligo)saccharides and glycoproteins were studied by haemagglutination-inhibition. Gel filtration shows that the lectin exists as a dimer above pH 5 and as a monomer below pH 3.5. This is confirmed by studies on the release of lectin subunits that were adsorbed from solution to lectin monomers immobilized onto Eupergit-c. From the gel filtration patterns it is calculated that a residue with pK(a) of about 4.4 is involved in dimer dissociation. Titration of glutamic acids (E60, E209) is postulated to be involved. CD spectroscopy shows that the secondary structure of the lectin is unchanged between pH 1 and 12.5, and that the tertiary structure remains unchanged between pH 5 and 12. In the acid pH region, reversible spectral changes occur that may be due to the titration of one or more amino acids with a pK(a) value of 3.9-4.2, probably aspartic acid. These residues are implicated in sugar-binding but not in dimerization of the lectin. Only at pH 12.5, irreversible denaturation occurs. Mukwa lectin displays full carbohydrate-binding capacity between pH 4 and 12, as is concluded from ELLA (Enzyme Linked Lectin Assay) using ovalbumin and fetuin, and from binding of the same glycoproteins to immobilized lectin monomers. The lectin is rapidly and fully reversibly demetallized at pH 2.5 with 5 mM EDTA. The demetallized lectin is completely devoid of sugar-binding activity. Mukwa lectin is a very thermostable molecule (at least till 85 degrees C). However, addition of non-ionic detergents substantially lowers its thermostability. PMID:19508209

  2. Isolation of an immunosuppressive lectin from Phaseolus vulgaris L. cv Cacahuate using stroma.

    Science.gov (United States)

    Vargas-Albores, F; Hernández, J; Córdoba, F; Zenteno, E

    1993-11-01

    An immunosuppressive lectin was isolated from seed of Phaseolus vulgaris cv Cacahuate using physically entrapped stroma. The lectin was found to be a 94 kDa tetrameric protein. When 50 micrograms, of this lectin were administered intraperitoneally 2 days before the immunization with sheep red blood cells, humoral response against the immunogen was completely inhibited. Other properties of the protein are discussed. PMID:8248029

  3. Isolation and properties of a lectin from the seeds of Mimosa invisa L

    OpenAIRE

    Chandrika, R; Shaila, MS

    1987-01-01

    A lectin has been purified from the seeds of Mimosa invisa L. by gel filtration and preparative Polyacrylamide gel electrophoresis. The purified lectin was homogeneous as judged by analytical Polyacrylamide gel electrophoresis, immunodiffusion and Immunoelectrophoresis. The apparent molecular weight is 100,000; the protein is a tetramer with two types of subunits (molecular weight 35,000 and 15,000). The lectin is a glycoprotein with approximately 21% carbohydrate and interacts with Sephadex ...

  4. Fucose-binding Lotus tetragonolobus lectin binds to human polymorphonuclear leukocytes and induces a chemotactic response.

    Science.gov (United States)

    VanEpps, D E; Tung, K S

    1977-09-01

    Fucose-binding L. tetragonolobus lectin to the surface of human polymorphonuclear leukocytes (PMN) and induces a chemotactic response. Both surface binding and chemotaxis are inhibited by free fucose but not by fructose, mannose, or galactose. The lectin-binding sites on PMN are unrelated to the A, B, or O blood group antigen. Utilization of this lectin should be a useful tool in isolating PMN membrane components and in analyzing the mechanism of neutrophil chemotaxis. PMID:330752

  5. Large Scale Magnetic Separation of Solanum tuberosum Tuber Lectin from Potato Starch Waste Water

    Science.gov (United States)

    Safarik, Ivo; Horska, Katerina; Martinez, Lluis M.; Safarikova, Mirka

    2010-12-01

    A simple procedure for large scale isolation of Solanum tuberosum tuber lectin from potato starch industry waste water has been developed. The procedure employed magnetic chitosan microparticles as an affinity adsorbent. Magnetic separation was performed in a flow-through magnetic separation system. The adsorbed lectin was eluted with glycine/HCl buffer, pH 2.2. The specific activity of separated lectin increased approximately 27 times during the isolation process.

  6. A simple fibril and lectin model for cyst walls of Entamoeba and perhaps Giardia

    OpenAIRE

    Samuelson, John; Robbins, Phillips

    2011-01-01

    Cyst walls of Entamoeba and Giardia protect them from environmental insults, stomach acids, and intestinal proteases. Each cyst wall contains a sugar homopolymer: chitin in Entamoeba and a unique N-acetylgalactosamine (GalNAc) homopolymer in Giardia. Entamoeba cyst wall proteins include Jacob lectins (carbohydrate-binding proteins) that cross-link chitin, chitinases that degrade chitin, and Jessie lectins that make walls impermeable. Giardia cyst wall proteins are also lectins that bind fibri...

  7. Entamoeba histolytica Lectins Contain Unique 6-Cys or 8-Cys Chitin-Binding Domains

    OpenAIRE

    Van Dellen, Katrina; Ghosh, Sudip K.; Robbins, Phillips W.; Loftus, Brendan; Samuelson, John

    2002-01-01

    The Jacob lectin, the most abundant glycoprotein in the cyst wall of Entamoeba invadens, contains five unique 6-Cys chitin-binding domains (CBDs). We identified Entamoeba histolytica and Entamoeba dispar genes encoding Jacob homologues, each of which contains two predicted 6-Cys CBDs. A unique 8-Cys CBD was found at the N termini of the E. histolytica chitinase and three other predicted lectins, called Jessie 1 to Jessie 3. The CBDs of four E. histolytica lectins (Jacob, chitinase, and Jessie...

  8. Circulating levels of interleukin-6, vascular endothelial growth factor, YKL-40, matrix metalloproteinase-3, and total aggrecan in spondyloarthritis patients during 3 years of treatment with TNFα inhibitors

    DEFF Research Database (Denmark)

    Pedersen, Susanne Juhl; Hetland, Merete Lund; Sørensen, Inge Juul; Østergaard, Mikkel; Nielsen, Hans Jørgen; Johansen, Julia Sidenius

    2010-01-01

    The objectives of the study were to investigate short and long-term changes and relations to treatment response of plasma interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), YKL-40, matrix metalloproteinase-3 (MMP-3), and total aggrecan in patients with spondyloarthritis (SpA) treated...... with tumor necrosis factor-alpha (TNFα) inhibitors and to compare with levels in healthy subjects. Biomarkers were measured in an observational cohort of 49 SpA patients (ankylosing spondylitis, n=32, and psoriatic arthritis, n=17) initiating TNFα inhibitor therapy (infliximab, n=38; etanercept, n=8...

  9. Circulating levels of interleukin-6, vascular endothelial growth factor, YKL-40, matrix metalloproteinase-3, and total aggrecan in spondyloarthritis patients during 3 years of treatment with TNF alpha inhibitors

    DEFF Research Database (Denmark)

    Pedersen, S.J.; Hetland, M.L.; Sørensen, Inge Juul; Østergaard, Mikkel; Nielsen, H.J.; Johansen, J.S.

    2010-01-01

    The objectives of the study were to investigate short and long-term changes and relations to treatment response of plasma interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), YKL-40, matrix metalloproteinase-3 (MMP-3), and total aggrecan in patients with spondyloarthritis (SpA) treated...... with tumor necrosis factor-alpha (TNF alpha) inhibitors and to compare with levels in healthy subjects. Biomarkers were measured in an observational cohort of 49 SpA patients (ankylosing spondylitis, n = 32, and psoriatic arthritis, n = 17) initiating TNF alpha inhibitor therapy (infliximab, n = 38...

  10. Circulating levels of interleukin-6, vascular endothelial growth factor, YKL-40, matrix metalloproteinase-3, and total aggrecan in spondyloarthritis patients during 3 years of treatment with TNFα inhibitors

    DEFF Research Database (Denmark)

    Sørensen, Inge Juul; Ostergaard, Mikkel; Nielsen, Hans Jørgen; Pedersen, Susanne Juhl; Hetland, Merete Lund; Østergaard, Mikkel; Johansen, Julia Sidenius

    2010-01-01

    The objectives of the study were to investigate short and long-term changes and relations to treatment response of plasma interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), YKL-40, matrix metalloproteinase-3 (MMP-3), and total aggrecan in patients with spondyloarthritis (SpA) treated...... with tumor necrosis factor-alpha (TNFa) inhibitors and to compare with levels in healthy subjects. Biomarkers were measured in an observational cohort of 49 SpA patients (ankylosing spondylitis, n=32, and psoriatic arthritis, n=17) initiating TNFa inhibitor therapy (infliximab, n=38; etanercept, n=8...

  11. Sulphation heterogeneity in the trisaccharide (GalNAcSbeta1, 4GlcAbeta1,3GalNAcS) isolated from the non-reducing terminal of human aggrecan chondroitin sulphate.

    Science.gov (United States)

    West, L A; Roughley, P; Nelson, F R; Plaas, A H

    1999-08-15

    We report here the isolation and sulphation isomer analyses of trisaccharides GalNAcS(beta1,4)GlcA(beta1,3)GalNAcS (in which S indicates sulphate) derived from the non-reducing termini of aggrecan chondroitin sulphate. Rat chondrosarcoma and human aggrecans were digested for 1 h at 37 degrees C with 30 micro-units of endo-chondroitinase ABC per microgram of chondroitin sulphate, and trisaccharides were isolated from the digests by ToyoPearl HW40S gel-filtration chromatography. Four trisaccharide species were identified; their sulphation isomer compositions, as determined by digestion with chondroitinase ACII and fluorescence-based ion-exchange HPLC, were GalNAc4Sbeta1,4GlcAbeta1,3GalNAc4S, GalNAc4Sbeta1,4GlcAbeta1,3GalNAc6S, GalNAc4,6Sbeta1,4GlcAbeta1, 3GalNAc4S and GalNAc4,6Sbeta1,4GlcAbeta1,3GalNAc6S. The abundances of such sequences in chondroitin sulphate on aggrecan from normal (foetal to 72 years of age) and from osteoarthritic human knee cartilages were also established. The results showed that non-reducing terminal GalNAc4S or GalNAc4,6S can be linked to either a 4-sulphated or a 6-sulphated disaccharide, suggesting that the sulphation of the last disaccharide might not have a direct effect on the specificity of chondroitin sulphate terminal GalNAc sulphotransferases. Furthermore, for each aggrecan preparation examined, the 4S-to-6S ratio of all chain interior disaccharides was equivalent to that in the last repeating disaccharides at the non-reducing terminus, suggesting that neither chondroitin 4-sulphotransferase nor chondroitin 6-sulphotransferase shows preferential activity near the chain terminus. PMID:10432320

  12. [Studies on the location of eight lectins in breast carcinoma].

    Science.gov (United States)

    Zheng, Z; Ji, Z M

    1990-12-01

    100 cases of breast carcinoma were studied with lectin affinitive histochemistry technology. The result showed that Ricinus comunis agglutinin (RCA1) was located in almost all intraductal carcinomas but one, while the positive rates in the other types were obviously low (P less than 0.05). The positive rate of Ulex europaeus agglutinin-I (UEA1) in well-differentiated types was higher than that in poorly-differentiated ones (P less than 0.05). The location of Peanut agglutinin (PNA), Bandeiraea Simplicifolia (BSL) and UEA1 in breast carcinomas exhibited some regularity and it might be useful in understanding the differentiation of breast carcinomas. No relationship between changes of the eight lectins and metastases in axillary lymph nodes was observed, but the authors considered that PNA-affinitive histochemistry was beneficial to the detection of micrometastases in lymph nodes. PMID:1964401

  13. How a plant lectin recognizes high mannose oligosaccharides.

    Science.gov (United States)

    Garcia-Pino, Abel; Buts, Lieven; Wyns, Lode; Imberty, Anne; Loris, Remy

    2007-08-01

    The crystal structure of Pterocarpus angolensis seed lectin is presented in complex with a series of high mannose (Man) oligosaccharides ranging from Man-5 to Man-9. Despite that several of the nine Man residues of Man-9 have the potential to bind in the monosaccharide-binding site, all oligomannoses are bound in the same unique way, employing the tetrasaccharide sequence Manalpha(1-2)Manalpha(1-6)[Manalpha(1-3)]Manalpha(1-. Isothermal titration calorimetry titration experiments using Man-5, Man-9, and the Man-9-containing glycoprotein soybean (Glycine max) agglutinin as ligands confirm the monovalence of Man-9 and show a 4-times higher affinity for Man-9 when it is presented to P. angolensis seed lectin in a glycoprotein context. PMID:17556509

  14. Properties of volkensin, a toxic lectin from Adenia volkensii.

    Science.gov (United States)

    Stirpe, F; Barbieri, L; Abbondanza, A; Falasca, A I; Brown, A N; Sandvig, K; Olsnes, S; Pihl, A

    1985-11-25

    Volkensin, a highly toxic protein from the roots of Adenia volkensii (kilyambiti, kinoria), was purified by affinity chromatography on acid-treated Sepharose 6B. The toxin is a glycoprotein (Mr 62,000, neutral sugar content 5.74%) consisting of an A subunit (Mr 29,000) and of a B subunit (Mr 36,000) linked by disulfide and noncovalent bond(s). The amino acid, amino sugar, and neutral sugar composition of the protein were determined. Volkensin is a galactose-specific lectin and is a potent inhibitor of eukaryotic protein synthesis in whole cells as well as in a cell-free system (a rabbit reticulocyte lysate). The inhibitory and the lectin activities are functions of the A and B subunits, respectively. Volkensin can be included amongst the ricin-like toxins and resembles most closely modeccin, the toxin of Adenia digitata. PMID:3932357

  15. Disseminated zygomycosis associated with erythroleukaemia: confirmation by lectin stains.

    OpenAIRE

    Benbow, E W; Delamore, I W; Stoddart, R. W.; Reid, H

    1985-01-01

    Zygomycosis is not often diagnosed in the United Kingdom, and so the possible importance of the findings in a patient with disseminated zygomycosis who had been treated with chemotherapy for erythroleukaemia was not appreciated until histological examination of specimens obtained at necropsy provided a presumptive diagnosis. No attempt had therefore been made to identify the organism by culture, and lectin binding methods were used to try to compensate for this. The characteristics of the hyp...

  16. Carbohydrate recognition by the antiviral lectin cyanovirin-N

    OpenAIRE

    Fujimoto, Yukiji K.; Green, David F.

    2012-01-01

    Cyanovirin-N is a cyanobacterial lectin with potent antiviral activity, and has been the focus of extensive pre-clinical investigation as a potential prophylactic for the prevention of the sexual transmission of the human immunodeficiency virus (HIV). Here we present a detailed analysis of carbohydrate recognition by this important protein, using a combination of computational methods, including extensive molecular dynamics simulations and Molecular-Mechanics/ Poisson–Boltzmann/Surface-Area (...

  17. Identification and characterisation of lectins in several tick species

    Czech Academy of Sciences Publication Activity Database

    Štěrba, J.; Dupejová, J.; Fišer, M.; Golovchenko, Maryna; Rudenko, Natalia; Grubhoffer, L.

    2009-01-01

    Roč. 276, S1 (2009), s. 88-88. ISSN 1742-464X. [34th FEBS Congress: Life's Molecular Interactions. 04.07.2009-09.07.2009, Prague] R&D Projects: GA AV ČR KJB600960906; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : lectins * hemagglutination acitvity * fibrinogen- related domain Subject RIV: CE - Biochemistry

  18. The Aging Heart, Myocardial Fibrosis and its Relationship to Circulating C-Type Natriuretic Peptide

    OpenAIRE

    Sangaralingham, S. Jeson; Huntley, Brenda K.; Martin, Fernando L; McKie, Paul M.; Bellavia, Diego; Ichiki, Tomoko; Harders, Gerald E.; Chen, Horng H.; Burnett, John C.

    2010-01-01

    Myocardial aging is characterized by LV fibrosis leading to diastolic and systolic dysfunction. Studies have established the potent anti-fibrotic and anti-proliferative properties of C-type natriuretic peptide (CNP), however the relationship between circulating CNP, LV fibrosis and associated changes in LV function with natural aging are undefined. Accordingly, we characterized the relationship of plasma CNP with LV fibrosis and function in 2, 11 and 20 month old male Fischer rats. Further in...

  19. Characterization of a new lectin involved in the protoplast regeneration of Bryopsis hypnoides

    Science.gov (United States)

    Niu, Jianfeng; Wang, Guangce; Lü, Fang; Zhou, Baicheng; Peng, Guang

    2009-09-01

    A group of coenocytic marine algae differs from higher plants, whose totipotency depends on an intact cell (or protoplast). Instead, this alga is able to aggregate its extruded protoplasm in sea water and generate new mature individuals. It is thought that lectins play a key role in the aggregation process. We purified a lectin associated with the aggregation of cell organelles in Bryopsis hypnoides. The lectin was ca. 27 kDa with a pI between pH 5 and pH 6. The absence of carbohydrate suggested that the lectin was not a glycoprotein. The hemagglutinating activity (HA) of the lectin was not dependent on the presence of divalent cations and was inhibited by N-Acetylgalactosamine, N-Acetylglucosamine, and the glycoprotein bovine submaxillary mucin. The lectin preferentially agglutinated Gram-negative bacterium. The HA of this lectin was stable between pH 4 to pH 10. Cell organelles outside the cytoplasm were agglutinated by the addition of lectin solution (0.5 mg ml-1). Our results suggest that the regeneration of B. hypnoides is mediated by this lectin. We also demonstrated that the formation of cell organelle aggregates was inhibited by nigericin in natural seawater (pH 8.0). Given that nigericin dissipates proton gradients across the membrane, we hypothesize that the aggregation of cell organelles was proton-gradient dependent.

  20. Characterization of a new lectin involved in the protoplast regeneration of Bryopsis hypnoides

    Institute of Scientific and Technical Information of China (English)

    NIU Jianfeng; WANG Guangce; L(U) Fang; ZHOU Baicheng; PENG Guang

    2009-01-01

    A group of coenocytic marine algae differs from higher plants, whose totipotency depends on an intact cell (or protoplast). Instead, this alga is able to aggregate its extruded protoplasm in sea water and generate new mature individuals. It is thought that lectins play a key role in the aggregation process. We purified a lectin associated with the aggregation of cell organelles in Bryopsis hypnoides. The lectin was ca. 27 kDa with a pI between pH 5 and pH 6. The absence of carbohydrate suggested that the lectin was not a glycoprotein. The hemagglutinating activity (HA) of the lectin was not dependent on the presence of divalent cations and was inhibited by N-Acetylgalactosamine, N-Acetylglucosamine, and the glycoprotein bovine submaxillary mucin. The lectin preferentially agglutinated Gram-negative bacterium. The HA of this lectin was stable between pH 4 to pH 10. Cell organelles outside the cytoplasm were agglutinated by the addition of lectin solution (0.5 mg ml-1). Our results suggest that the regeneration of B. hypnoides is mediated by this lectin. We also demonstrated that the formation of cell organelle aggregates was inhibited by nigericin in natural seawater (pH 8.0). Given that nigericin dissipates proton gradients across the membrane, we hypothesize that the aggregation of cell organelles was proton-gradient dependent.

  1. Microencapsulation of lectin anti-cancer agent and controlled release by alginate beads, biosafety approach.

    Science.gov (United States)

    El-Aassar, M R; Hafez, Elsayed E; El-Deeb, Nehal M; Fouda, Moustafa M G

    2014-08-01

    Hepatocellular carcinoma (HCC) is considered as one of the most aggressive cancer worldwide. In Egypt, the prevalence of HCC is increasing during last years. Recently, drug-loaded microparticles were used to improve the efficiency of various medical treatments. This study is designed to evaluate the anticancer potentialities of lectins against HCC while hinting to its safety usage. The aim is also extended to encapsulate lectins in alginate microbeads for oral drug delivery purposes. The extracted lectins showed anti-proliferative effect against HCC with a percentage of 60.76% by using its nontoxic dose with an up-regulation of P53 gene expression. Concerning the handling of lectin alginate microbeads for oral drug delivery, the prepared lectin alginate beads were ∼100μm in diameter. The efficiency of the microcapsules was checked by scanning electron microscopy, the SEM showed the change on the alginate beads surface revealing the successful lectin encapsulation. The release of lectins from the microbeads depended on a variety of factors as the microbeads forming carriers and the amount-encapsulated lectins. The Pisum sativum extracted lectins may be considered as a promising agent in controlling HCC and this solid dosage form could be suitable for oral administration complemented with/or without the standard HCC drugs. PMID:24857870

  2. Sialic acid specific lectins from Episesarma tetragonum (Decapoda, Grapsidae): isolation, purification and characterization

    OpenAIRE

    R. Viswambari Devi; M.R. Basil Rose; P.D. Mercy

    2013-01-01

    Two sialic acid specific lectins Episesarma tetragonum agglutinin–1 and 2 were purified from the hemolymph of the Mangrove crab, Episesarma tetragonum. The major lectin was purified using CNBr-activated sepharose 4B conjugated to fetuin. N-acetyl glucosamine containing buffer was used for elution. The hemagglutination activity of purified lectin was inhibited by glycoproteins containing Siaα, 2-3Galβ, 1-4 GlcNAc linkages. On SDS-PAGE, the molecular weight of calcium dependent lectin was obser...

  3. Purification and partial characterization of a mitogenic lectin from the latex of Euphorbia marginata.

    Science.gov (United States)

    Stirpe, F; Licastro, F; Morini, M C; Parente, A; Savino, G; Abbondanza, A; Bolognesi, A; Falasca, A I; Rossi, C A

    1993-08-20

    A lectin was purified from the latex of Euphorbia marginata by affinity chromatography on acid-treated Sepharose 6B and elution with lactose. The lectin is a glycoprotein composed of two identical subunits with M(r) 30,000, approx. The haemagglutinating activity of the lectin is not specific for any human blood group, and is inhibited by galactose and galactose-containing sugars and by gentiobiose. The lectin is strongly mitogenic for human T-lymphocytes and induces the release of interleukin-1 beta and tumor necrosis factor-alpha from cultured mononuclear cells. PMID:8353129

  4. Prenatal diagnosis of methymalonic aciduria and homocystinuria cblC type using DNA analysis

    Directory of Open Access Journals (Sweden)

    Antonietta Zappu

    2015-12-01

    Full Text Available Methylmalonic aciduria (MMA and homocystinuria, cblC type is the most frequent inborn error of vitamin B12. CblC patients present with a heterogeneous clinical picture.To date, the early prenatal diagnosis of MMA and homocystinuria, cblC type is performed by determination of methylmalonic acid and total homocysteine (Hcy in amniotic fluid supernatant. In this paper we report a case of prenatal diagnosis, using genetic analysis, of MMA and homocystinuria, cblC type in an at risk couple. Direct sequencing analysis of the amplified products of chorionic villi biopsy extracted DNA showed normal sequence in the fetal DNA. Mutation analysis of the MMACHC gene is more cost-effective and less time-consuming than the biochemical approach. Early prenatal treatment may have an impact on the long-term complications associated with cblC disease. Future studies with the aim of determining the long-term benefits of daily parenteral OHCbl started soon after conception in at risk mothers should be considered. In this context early prenatal diagnosis could determine whether therapy needs to be continued.

  5. Regional differences in lectin binding patterns of vestibular hair cells

    Science.gov (United States)

    Baird, Richard A.; Schuff, N. R.; Bancroft, J.

    1994-01-01

    Surface glycoconjugates of hair cells and supporting cells in the vestibular endorgans of the bullfrog were identified using biotinylated lectins with different carbohydrate specificities. Lectin binding in hair cells was consistent with the presence of glucose and mannose (CON A), galactose (RCA-I), N-acetylgalactosamine (VVA), but not fucose (UEA-I) residues. Hair cells in the bullfrog sacculus, unlike those in the utriculus and semicircular canals, did not stain for N-acetylglucosamine (WGA) or N-acetylgalactosamine (VVA). By contrast, WGA and, to a lesser extent, VVA, differentially stained utricular and semicircular canal hair cells, labeling hair cells located in peripheral, but not central, regions. In mammals, WGA uniformly labeled Type 1 hair cells while labeling, as in the bullfrog, Type 2 hair cells only in peripheral regions. These regional variations were retained after enzymatic digestion. We conclude that vestibular hair cells differ in their surface glycoconjugates and that differences in lectin binding patterns can be used to identify hair cell types and to infer the epithelial origin of isolated vestibular hair cells.

  6. Labeling of lectin receptors during the cell cycle.

    Science.gov (United States)

    Garrido, J

    1976-12-01

    Labeling of lectin receptors during the cell cycle. (Localizabión de receptores para lectinas durante el ciclo celular). Arch. Biol. Med. Exper. 10: 100-104, 1976. The topographic distribution of specific cell surface receptors for concanavalin A and wheat germ agglutinin was studied by ultrastructural labeling in the course of the cell cycle. C12TSV5 cells were synchronized by double thymidine block or mechanical selection (shakeoff). They were labeled by means of lectin-peroxidase techniques while in G1 S, G2 and M phases of the cycle. The results obtained were similar for both lectins employed. Interphase cells (G1 S, G2) present a stlihtly discontinous labeling pattern that is similar to the one observed on unsynchronized cells of the same line. Cells in mitosis, on the contrary, present a highly discontinous distribution of reaction product. This pattern disappears after the cells enters G1 and is not present on mitotic cells fixed in aldehyde prior to labeling. PMID:1030938

  7. Crystal structure of a symbiosis-related lectin from octocoral.

    Science.gov (United States)

    Kita, Akiko; Jimbo, Mitsuru; Sakai, Ryuichi; Morimoto, Yukio; Miki, Kunio

    2015-09-01

    D-Galactose-binding lectin from the octocoral, Sinularia lochmodes (SLL-2), distributes densely on the cell surface of microalgae, Symbiodinium sp., an endosymbiotic dinoflagellate of the coral, and is also shown to be a chemical cue that transforms dinoflagellate into a non-motile (coccoid) symbiotic state. SLL-2 binds with high affinity to the Forssman antigen (N-acetylgalactosamine(GalNAc)α1-3GalNAcβ1-3Galα1-4Galβ1-4Glc-ceramide), and the presence of Forssman antigen-like sugar on the surface of Symbiodinium CS-156 cells was previously confirmed. Here we report the crystal structures of SLL-2 and its GalNAc complex as the first crystal structures of a lectin involved in the symbiosis between coral and dinoflagellate. N-Linked sugar chains and a galactose derivative binding site common to H-type lectins were observed in each monomer of the hexameric SLL-2 crystal structure. In addition, unique sugar-binding site-like regions were identified at the top and bottom of the hexameric SLL-2 structure. These structural features suggest a possible binding mode between SLL-2 and Forssman antigen-like pentasaccharide. PMID:26022515

  8. Polymorphisms in the Mannose-Binding Lectin Gene are Associated with Defective Mannose-Binding Lectin Functional Activity in Crohn's Disease Patients.

    Science.gov (United States)

    Choteau, Laura; Vasseur, Francis; Lepretre, Frederic; Figeac, Martin; Gower-Rousseau, Corine; Dubuquoy, Laurent; Poulain, Daniel; Colombel, Jean-Frederic; Sendid, Boualem; Jawhara, Samir

    2016-01-01

    Mannose-binding lectin, together with mannose-associated serine proteases, activates the lectin pathway of the complement system and subsequent inflammatory mechanisms. An association between mannose-binding lectin deficiency and anti-Saccharomyces cerevisiae antibody levels is observed in Crohn's disease and this deficiency is frequently associated with a severe Crohn's disease phenotype. In the present study, we assessed the relationship between serum concentrations of mannose-binding lectin, mannose-binding lectin functional activity, MBL2 and NOD2 polymorphisms, anti-S. cerevisiae antibody levels and clinical Crohn's disease phenotype in 69 Crohn's disease patients and 30 age- and sex-matched healthy controls. The results show that the MBL2 variant rs5030737 at codon 52 was associated with a low level of mannose-binding lectin and impaired mannose-binding lectin-mannose-associated serine protease (MBL-MASP) functional activity in Crohn's disease patients. This MBL2 variant was also associated with a higher level of anti-S. cerevisiae antibodies. In addition, the NOD2 variant rs2066844, which is associated with susceptibility to Crohn's disease, was significantly correlated with an impairment in MBL-MASP functional activity. These results provide evidence that Crohn's disease patients have an impairment in MBL-MASP functional activity and that this defect is associated with MBL2 and NOD2 variants. PMID:27404661

  9. Effect of the Lectin of Bauhinia variegata and Its Recombinant Isoform on Surgically Induced Skin Wounds in a Murine Model

    OpenAIRE

    Rodrigo Bainy Leal; Valdemiro Amaro da Silva Júnior; Benildo Sousa Cavada; Edson Holanda Teixeira; Francisco Vassiliepe Sousa Arruda; Ana Lúcia Figueiredo Porto; Victor Alves Carneiro; Mayron Alves de Vasconcelos; Francisco Flávio Vasconcelos Evaristo; Rafaela Mesquita Bastos; Luiz Gonzaga do Nascimento Neto; Luciano da Silva Pinto

    2011-01-01

    Lectins are a structurally heterogeneous group of highly specific carbohydrate-binding proteins. Due to their great biotechnological potential, lectins are widely used in biomedical research. The purpose of the present study was to evaluate the healing potential of the lectin of Bauhinia variegata (nBVL) and its recombinant isoform (rBVL-1). Following surgical creation of dorsal skin wounds, seven groups of mice were submitted to topical treatment for 12 days with lectin, D-galactose, BSA and...

  10. Preparation and identification of 1.3 copies C-type HBV transgenic mice

    Directory of Open Access Journals (Sweden)

    Mei-juan CHEN

    2011-09-01

    Full Text Available Objective To prepare 1.3 copies C-type HBV transgenic mice for providing a better model for the prevention and treatment of hepatitis B.Methods The HBV transgenic mice were generated by microinjection of 1.3 copies C-type HBV genome into the pronucleus of FVB /N zygotes.PCR,ELISA,RT-PCR and immunohistochemistry were used to detect the integration,replication and expression of HBV gene in the transgenic mice.Results Tow thousand two hundred and eighty-two fertilized eggs were injected and a total of 2024 survived.The survival rate of injection was 88.7%.The injected eggs were transplanted into 72 pseudo pregnant female mice,among which 59 became pregnant.The pregnancy rate was 81.9%.One hundred and eighty-five F0 offsprings were produced with 19 positive mice as detected by PCR,and the positive rate was 10.3%.RT-PCR revealed that HBV DNA replication of 102-103 copies/ml existed in serum of 6 mice.Ninety-six F1 offsprings were produced,of which 33 were positive for HBV DNA replication as detected by PCR,the positive rate was 34.4%.RT-PCR showed that HBV DNA replication was observed in 10 mice with 102-103 copies/ml.Three mice were randomly chosen from each of F0 and F1 generations to detect the HBsAg expression in livers and kidneys by immunohistochemistry.The results showed that HBsAg expressed in both livers and kidneys,and it was stronger in kidneys than in livers.Conclusion The 1.3 copies C-type HBV gene can not only replicate and express in the transgenic mice produced,but it also can be transmitted to the next generation of these mice.

  11. Processing-independent analysis for pro-C-type natriuretic peptide

    DEFF Research Database (Denmark)

    Lippert, Solvej Kølvraa; Rehfeld, Jens F.; Gøtze, Jens Peter

    2010-01-01

    C-type natriuretic peptide (CNP) is expressed in several human tissues. We designed a specific processing-independent assay for proCNP-derived products and quantitated the concentrations in human seminal plasma from normal and vasectomized men. Antibodies were raised against the N-terminus of human...... proCNP 11-27. Samples were incubated with trypsin prior to immunoassay, which allows for the measurement of "total" proCNP irrespective of the degree of post-translational processing. Seminal plasma from normal young men and vasectomized men were collected and quantitated; the molecular heterogeneity...

  12. [Purification of lectin from perch (Persa fluviatilis L.) roe specific to cellobiose and study of its characteristics].

    Science.gov (United States)

    Antoniuk, V O

    2004-01-01

    Two lectins with different carbohydrate specificity were purified from perch (Persa fluviatilis L.) roe (coastal ecological form) by affinity chromatography on ovariomucine H-sepharose from a human ovary cyst. One lectin was eluted by cellobiose and another lectin was eluted by L-fucose. The L-fucose-specific lectin interacted only with L-fucose and its derivatives, but did not interact with cellobiose and salicin. The cellobiose-specific lectin interacted with all the examined carbohydrates, but cellobiose was the best inhibitor. This lectin can be also purified on cellulose as an affinity sorbent. Unlike the L-fucose-specific lectin from perch roe, the cellobiose-specific lectin is less soluble in water-saline solutions. Lectin solubility increases greatly in presence of specific inhibitors, cellobiose, in particular. L-fucose, alpha-methyl-L-fucopyranoside and 4-nitrophenyl-alpha-L-fucopyranoside are equivalent inhibitors for both lectins. According to SDS-PAGE data, the lectins contain two components with molecular weight 12-13 kDa. In solutions, these components form molecules with 50 or 100 kDa (depending on pH). Data obtained from electrophoresis in PAAG in alkaline (pH 8.9) and acidic system (pH 4.3), and SDS-PAGE did not display essential distinctions between these both lectins. PMID:15909420

  13. [Comparative lectin histochemical analysis of the duodenal glands in various mammals].

    Science.gov (United States)

    Iatskovskiĭ, A N; Lutsik, A D

    1991-02-01

    Composition and histotopography of lectin receptors have been studied in 12 species of mammals with various nutritional specialization: carnivorous, phytophagous and omnivorous. In cells of the duodenal glands of the carnivorous and omnivorous receptors to concanavalin A and lentil lectin (D-mannosoglycans ) are absent and they are present in the glands of the phytophagous animals. In cells of some parts of the glands presence of receptors to soya bean lectin (N-acetyl-D-galactosamine++) is the most characteristic sign of the duodenal glands in the carnivorous and phytophagous animals. Together with certain differences, depending on the nutritional way of the animals, specific peculiarities of lectins binding with glandulocytes of the duodenal glands are demonstrated. The data on rearrangement of the lectin receptors are obtained during the process of cellular differentiation. Presence of N-acetyl-D-galactosamine++ remnants-biding soya bean lectin in composition of oligosaccharide++ chains of glycoconjugates is a sign of low differential degree of the glandular cells. In more differentiated cells concealment in oligosaccharide chains of D-galactose remnants (peanut and castor-oil lectins receptors) by L-fucose, N-acetil-D-glucosamin remnants and sialic acid can have place; this is demonstrated as accumulation of receptors to wheat germ and Laburnum anagyroides lectins in the glandular cells. PMID:2053882

  14. Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity

    DEFF Research Database (Denmark)

    Pedersen, Johannes W; Bennett, Eric P; Schjoldager, Katrine T-B G; Meldal, Morten; Holmér, Andreas P; Blixt, Ola; Cló, Emiliano; Levery, Steven B; Clausen, Henrik; Wandall, Hans H

    2011-01-01

    sequences of mucins MUC1, MUC2, MUC4, MUC5AC, MUC6, and MUC7 as well as a random glycopeptide bead library, we examined the binding properties of four different lectin domains. The lectin domains of GalNAc-T1, -T2, -T3, and -T4 bound different subsets of small glycopeptides. These results indicate an...

  15. Serum levels, ontogeny and heritability of chicken mannan-binding lectin (MBL)

    DEFF Research Database (Denmark)

    Laursen, S.B.; Hedemand, J.E.; Nielsen, O.L.; Thiel, S.; Koch, C.; Jensenius, J.C.

    1998-01-01

    Mannan-binding lectin (MBL) is a serum lectin found in mammals and recently also in birds. It is thought to play an important role in the innate immune defence through binding to surface carbohydrates on micro-organisms followed by complement activation via the MBL pathway. This results in...

  16. Rapid bead-based immunoassay for measurement of mannose-binding lectin

    DEFF Research Database (Denmark)

    Bay, J T; Garred, P

    2009-01-01

    Mannose-binding lectin (MBL) is a serum protein, which functions as an opsonin and initiator of the lectin pathway of complement. The serum concentration of MBL shows great interindividual variation because of common polymorphisms in the MBL2 gene. Although several quantitative MBL immunoassays...

  17. Evolutionary conservation of mannan-binding lectin (MBL) in bony fish

    DEFF Research Database (Denmark)

    Kania, Per Walter; Sørensen, Rasmus Reng; Koch, Claus;

    2010-01-01

    to characterize members of the lectin pathway in fish. We identified and characterized three homologues of mannan-binding lectin (MBL) with a bona fide collectin structure. By means of RT-PCR and immunohistochemistry using monoclonal antibodies we found that they were synthesized in the spleen, the...

  18. Mannose-binding lectin variant alleles and the risk of arterial thrombosis in systemic lupus erythematosus

    DEFF Research Database (Denmark)

    Øhlenschlaeger, Tommy; Garred, Peter; Madsen, Hans O;

    2004-01-01

    Cardiovascular disease is an important complication in patients with systemic lupus erythematosus (SLE). Variant alleles of the mannose-binding lectin gene are associated with SLE as well as with severe atherosclerosis. We determined whether mannose-binding lectin variant alleles were associated...

  19. Mannose-binding lectin polymorphisms and susceptibility to infection in systemic lupus erythematosus

    DEFF Research Database (Denmark)

    Garred, P; Madsen, H O; Halberg, P;

    1999-01-01

    To determine whether variant alleles in the coding portion of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to systemic lupus erythematosus (SLE) and concomitant infections.......To determine whether variant alleles in the coding portion of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to systemic lupus erythematosus (SLE) and concomitant infections....

  20. Multiplicity of carbohydrate-binding sites in -prism fold lectins: occurrence and possible evolutionary implications

    Indian Academy of Sciences (India)

    Alok Sharma; Divya Chandran; Desh D Singh; M Vijayan

    2007-09-01

    The -prism II fold lectins of known structure, all from monocots, invariably have three carbohydrate-binding sites in each subunit/domain. Until recently, -prism I fold lectins of known structure were all from dicots and they exhibited one carbohydrate-binding site per subunit/domain. However, the recently determined structure of the -prism fold I lectin from banana, a monocot, has two very similar carbohydrate-binding sites. This prompted a detailed analysis of all the sequences appropriate for two-lectin folds and which carry one or more relevant carbohydrate-binding motifs. The very recent observation of a -prism I fold lectin, griffithsin, with three binding sites in each domain further confirmed the need for such an analysis. The analysis demonstrates substantial diversity in the number of binding sites unrelated to the taxonomical position of the plant source. However, the number of binding sites and the symmetry within the sequence exhibit reasonable correlation. The distribution of the two families of -prism fold lectins among plants and the number of binding sites in them, appear to suggest that both of them arose through successive gene duplication, fusion and divergent evolution of the same primitive carbohydrate-binding motif involving a Greek key. Analysis with sequences in individual Greek keys as independent units lends further support to this conclusion. It would seem that the preponderance of three carbohydrate-binding sites per domain in monocot lectins, particularly those with the -prism II fold, is related to the role of plant lectins in defence.

  1. Purification and partial characterization of a lectin from the seeds of Trichosanthes kirilowii Maximowicz.

    Science.gov (United States)

    Falasca, A I; Abbondanza, A; Barbieri, L; Bolognesi, A; Rossi, C A; Stirpe, F

    1989-03-27

    A lectin was purified from the seeds of Trichosanthes kirilowii, belonging to the family Cucurbitaceae, growing in China. The lectin is a glycoprotein of 57 kDa, consists of two subunits with apparent molecular masses of 37 and 25 kDa, is specific for galactose, and is not mitogenic for human lymphocytes. PMID:2707434

  2. The relationship between C-type natriuretic peptide and cognitive impairment in older patients with Type 2 diabetes

    International Nuclear Information System (INIS)

    Objective: To investigate the relationship between C-type natriuretic peptide and cognitive impairment in older patients with type 2 diabetes, and to explore the pathogenesis of diabetic cognitive impairment. Methods: According to the Montreal Cognitive Assessment (MoCA) scores, 80 type 2 diabetic patients over the age of 60 years were divided into two groups, one group including 31 cases with cognitive impairment, the other 49 patients with non-cognitive impairment. And 80 normal participants were selected as the control group. Plasma level of C-type natriuretic peptide was measured by radio-immunity assay in all subjects. The changes and associations of the plasma C-type natriuretic peptide level among three groups was analyzed. Result: In the non-cognitive impairment group, plasma level of C-type natriuretic peptide was higher than that in the control group (P<0.01). But the plasma level of C-type natriuretic peptide in the cognitive impairment group was degraded, significantly deferent with those in the control group and the non-cognitive impairment group (P<0.01). MoCA scores of the cognitive impairment group positively correlated with plasma level of C-type natriuretic peptide (r=0.513, P<0.01). Conclusion: In the early period of type 2 diabetes,the secretion of C-type natriuretic peptide was increased. When diabetic cognitive impairment complicated,the secretion of C-type natriuretic peptide was decompensated. Then plasma level of C-type natriuretic peptide become low. The level of C-type natriuretic peptide closely correlated with diabetic cognitive impairment. It was suggested that diabetic angiopathies may act an important role in the pathogenesis of diabetic cognitive impairment. (authors)

  3. Insecticidal Activity of Plant Lectins and Potential Application in Crop Protection

    Directory of Open Access Journals (Sweden)

    Maria Lígia R. Macedo

    2015-01-01

    Full Text Available Lectins constitute a complex group of proteins found in different organisms. These proteins constitute an important field for research, as their structural diversity and affinity for several carbohydrates makes them suitable for numerous biological applications. This review addresses the classification and insecticidal activities of plant lectins, providing an overview of the applicability of these proteins in crop protection. The likely target sites in insect tissues, the mode of action of these proteins, as well as the use of lectins as biotechnological tools for pest control are also described. The use of initial bioassays employing artificial diets has led to the most recent advances in this field, such as plant breeding and the construction of fusion proteins, using lectins for targeting the delivery of toxins and to potentiate expected insecticide effects. Based on the data presented, we emphasize the contribution that plant lectins may make as tools for the development of integrated insect pest control strategies.

  4. PO-20 - Crosstalk between the lectin pathway and haemostasis in patients with pulmonary cancer

    DEFF Research Database (Denmark)

    Larsen, J B; Christensen, T D; Hvas, C L; Hvas, A M

    INTRODUCTION: Recent research has focused on the complement system in cancer, including the lectin pathway of complement activation. Mannose-binding lectin (MBL), a key activator of the lectin pathway, can bind to tumor cell surfaces in vitro, and lectin pathway activation is increased in several...... coagulation and impair fibrinolysis in vitro, but the significance of this in a clinical setting is not well understood. AIM: We aim to investigate associations between lectin pathway and haemostatic activation in patients with lung cancer undergoing thoracoscopic surgery. MATERIALS AND METHODS: Patients with...... lung cancer (n=60) eligible for thoracoscopic tumor resection were included as part of a randomized controlled trial, the COPPVATS project (EudraCT no: 2012-002409-23), conducted at the Department of Thoracic Surgery, Aarhus University Hospital. Exclusion criteria were arterial or venous thrombosis...

  5. A hypothesis on the origin of C-type asteroids and carbonaceous chondrites

    CERN Document Server

    Busarev, V V

    2012-01-01

    A hypothesis based on observational and theoretical results on the origin of C-type asteroids and carbonaceous chondrites is proposed. Asteroids of C-type and close BGF-types could form from hydrated silicate-organic matter accumulated in the cores of water-differentiated (due to 26Al and other short-lived isotopes decay) bodies existed in the growth zones of Jupiter. Gravitational scattering of such bodies by Jupiter at its final stage of formation to the main asteroid belt might have led to fragmentation and re-accretion of their primitive materials on the surfaces of many asteroids and/or asteroid parent bodies. The hypothesis makes clear a row of long-standing puzzling facts, the main of which are as follows. The low-albedo and carbonaceous-chondritic surface properties of (1) Ceres contradict to its probable differentiated structure and icy crust (e. g., Thomas et al., 2005, Nature 437: 224-226; Castillo-Rogez et al., 2010, Icarus 205, 443-459), but it could be explained by the process of primitive matte...

  6. Isolation and characterization of a c-type lysozyme from the nurse shark.

    Science.gov (United States)

    Hinds Vaughan, Nichole; Smith, Sylvia L

    2013-12-01

    Lysozyme is a ubiquitous antibacterial enzyme that occurs in numerous invertebrate and vertebrate species. Three forms have been described c-type, g-type and i-type which differ in primary structure. Shark lysozyme has not been characterized; here we report on the isolation and characterization of lysozyme from unstimulated shark (Ginglymostoma cirratum) leukocytes and provide amino acid sequence data across the highly conserved active site of the molecule identifying it to be a c-type lysozyme. A leukocyte lysate was applied either (a) to the first of two sequential DE-52 cellulose columns or alternatively, (b) to a DEAE-Sepharose column. Lysozyme activity in lysate and active fractions was identified by zones of lysis of Micrococcus lysodeikticus cell walls on lysoplates and zones of growth inhibition in agar diffusion assays using Planococcus citreus as the target organism. SDS-PAGE analysis revealed a 14 kDa protein which was identified as lysozyme by mass spectroscopic analysis of peptides, reactivity against anti-HEWL antibodies on a Western blot, hydrolysis of M. lysodeikticus cell walls, and inhibition of growth of P. citreus on AU-gel blots in which the area of growth inhibition correlated to a 14 kDa protein. PMID:24084042

  7. Mannose-specific lectin from the mushroom Hygrophorus russula.

    Science.gov (United States)

    Suzuki, Tomohiro; Sugiyama, Kozue; Hirai, Hirofumi; Ito, Hiroyuki; Morita, Tatsuya; Dohra, Hideo; Murata, Takeomi; Usui, Taichi; Tateno, Hiroaki; Hirabayashi, Jun; Kobayashi, Yuka; Kawagishi, Hirokazu

    2012-05-01

    A lectin was purified from the mushroom Hygrophorus russula by affinity chromatography on a Sephadex G-50 column and BioAssist S cation exchange chromatography and designated H. russula lectin (HRL). The results of sodium dodecyl sulfate-polyaclylamidegel electrophoresis, gel filtration and matrix-assisted laser desorption ionization time-of-flight mass spectrometry of HRL indicated that it was composed of four identical 18.5 kDa subunits with no S-S linkage. Isoelectric focusing of the lectin showed bands near pI 6.40. The complete sequence of 175 amino acid residues was determined by amino acid sequencing of intact or enzyme-digested HRL. The sequence showed homology with Grifola frondosa lectin. The cDNA of HRL was cloned from RNA extracted from the mushroom. The open reading frame of the cDNA consisted of 528 bp encoding 176 amino acids. In hemagglutination inhibition assay, α1-6 mannobiose was the strongest inhibitor and isomaltose, Glcα1-6Glc, was the second strongest one, among mono- and oligosaccharides tested. Frontal affinity chromatography indicated that HRL had the highest affinity for Manα1-6(Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc, and non-reducing terminal Manα1-6 was essential for the binding of HRL to carbohydrate chains. The sugar-binding specificity of HRL was also analyzed by using BIAcore. The result from the analysis exhibited positive correlations with that of the hemagglutination inhibition assay. All the results suggested that HRL recognized the α1-6 linkage of mannose and glucose, especially the Manα1-6 bond. HRL showed a mitogenic activity against spleen lymph cells of an F344 rat. Furthermore, an enzyme-linked immunosorbent assay showed strong binding of HRL to human immunodeficiency virus type-1 gp120. PMID:22198564

  8. Lectin-like receptors: Evaluation of interactions using glycodendrimers

    Czech Academy of Sciences Publication Activity Database

    Vannucci, Luca; Pospíšil, Miloslav; Rossmann, Pavel; Difato, Francesco; Křen, Vladimír; Luptovcová, Martina; Svoboda, Jan; Kuldová, Markéta; Bezouška, Karel; Saieh, M.; Fišerová, Anna

    Hersonissos : Blackwell Publishing, 2006, s. 201-201. [World Congress on Advances in Oncology /11./ and International Symposium on Molecular Medicine /9./. Hersonissos (GR), 12.10.2006-14.10.2006] R&D Projects: GA AV ČR IAA5020403; GA AV ČR IAA500200620; GA AV ČR IAA500200509; GA AV ČR IAA500200510 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50110509 Keywords : lectin-like receptors * glycodendrimers * ligands Subject RIV: EE - Microbiology, Virology

  9. Polymorphisms in the Mannose-Binding Lectin Gene are Associated with Defective Mannose-Binding Lectin Functional Activity in Crohn’s Disease Patients

    Science.gov (United States)

    Choteau, Laura; Vasseur, Francis; Lepretre, Frederic; Figeac, Martin; Gower-Rousseau, Corine; Dubuquoy, Laurent; Poulain, Daniel; Colombel, Jean-Frederic; Sendid, Boualem; Jawhara, Samir

    2016-01-01

    Mannose-binding lectin, together with mannose-associated serine proteases, activates the lectin pathway of the complement system and subsequent inflammatory mechanisms. An association between mannose-binding lectin deficiency and anti-Saccharomyces cerevisiae antibody levels is observed in Crohn’s disease and this deficiency is frequently associated with a severe Crohn’s disease phenotype. In the present study, we assessed the relationship between serum concentrations of mannose-binding lectin, mannose-binding lectin functional activity, MBL2 and NOD2 polymorphisms, anti-S. cerevisiae antibody levels and clinical Crohn’s disease phenotype in 69 Crohn’s disease patients and 30 age- and sex-matched healthy controls. The results show that the MBL2 variant rs5030737 at codon 52 was associated with a low level of mannose-binding lectin and impaired mannose-binding lectin–mannose-associated serine protease (MBL-MASP) functional activity in Crohn’s disease patients. This MBL2 variant was also associated with a higher level of anti-S. cerevisiae antibodies. In addition, the NOD2 variant rs2066844, which is associated with susceptibility to Crohn’s disease, was significantly correlated with an impairment in MBL-MASP functional activity. These results provide evidence that Crohn’s disease patients have an impairment in MBL-MASP functional activity and that this defect is associated with MBL2 and NOD2 variants. PMID:27404661

  10. Gal/GalNAc specific multiple lectins in marine bivalve Anadara granosa.

    Science.gov (United States)

    Adhya, Mausumi; Singha, Biswajit

    2016-03-01

    Complete lectin mapping of molluscs with their diversified recognition pattern and possible role in lectin-carbohydrate interaction based immune response triggering need much attention. In this communication, Gal/GalNAc specific three lectins AGL-IA (Anadara granosa lectin-IA), AGL-IB (A. granosa lectin-IB) and AGL-IV (A. granosa lectin-IV) and a lectin having hemolytic activity AGL-III (A. granosa lectin-III) were purified from the plasma of A. granosa bivalve by a combination of gel filtration and affinity chromatography. AGL-IA and IB were oligomeric lectins whereas, AGL-III and IV were monomeric. The molecular weight of AGL-IA, IB, III and IV were 375, 260, 45 and 33 kDa respectively. AGL-IA and IV agglutinated both rabbit and pronase treated human erythrocytes, whereas AGL-IB agglutinated only rabbit erythrocytes. AGL-III was found to agglutinate rabbit erythrocytes, however, it caused hemolysis of pronase treated human erythrocytes. The activity of all four lectins was calcium dependent and maximum at a pH range 7-8. Apart from Gal/GalNAc specific, the four lectins showed substantial differences in their carbohydrate recognition pattern. Moreover, there was a difference in the carbohydrate specificity between AGL-III and other three lectins (AGL-IA, AGL-IB and AGL-IV) towards polyvalent glycotope. On the one hand, 'cluster glycoside effect' i.e., an enhancement of the activity of a multivalent ligand, was observed for carbohydrate specificities of AGL-IA, AGL-IB, AGL-IV. On the other hand, the effect of multivalent ligands on the carbohydrate specificity of AGL-III was opposite of cluster glycoside effect. The affinity of AGL-IA, AGL-IB and AGL-IV for ligands can be ranked as follows: glycoproteins > polysaccharide > oligosaccharides and monosaccharides. However, Gal related monosaccharides were the best inhibitors of AGL-III and the inhibitory activity decreased gradually in the following order: monosaccharide > disaccharide > polysaccharide. Thus

  11. Collectin-11/MASP complex formation triggers activation of the lectin complement pathway--the fifth lectin pathway initiation complex

    DEFF Research Database (Denmark)

    Ma, Ying Jie; Skjoedt, Mikkel-Ole; Garred, Peter

    2013-01-01

    complement pathway regulator MAP-1. Furthermore, we found that complex formation between recombinant collectin-11 and recombinant MASP-2 on Candida albicans leads to deposition of C4b. Native collectin-11 in serum mediated complement activation and deposition of C4b and C3b, and formation of the terminal...... complement complex on C. albicans. Moreover, spiking collectin-11-depleted serum, which did not mediate complement activation, with recombinant collectin-11 restored the complement activation capability. These results define collectin-11 as the fifth recognition molecule in the lectin complement pathway in...

  12. Development and optimization of a competitive binding assay for the galactophilic low affinity lectin LecA from Pseudomonas aeruginosa.

    Science.gov (United States)

    Joachim, Ines; Rikker, Sebastian; Hauck, Dirk; Ponader, Daniela; Boden, Sophia; Sommer, Roman; Hartmann, Laura; Titz, Alexander

    2016-08-16

    Infections with the Gram-negative bacterium Pseudomonas aeruginosa result in a high mortality among immunocompromised patients and those with cystic fibrosis. The pathogen can switch from planktonic life to biofilms, and thereby shields itself against antibiotic treatment and host immune defense to establish chronic infections. The bacterial protein LecA, a C-type lectin, is a virulence factor and an integral component for biofilm formation. Inhibition of LecA with its carbohydrate ligands results in reduced biofilm mass, a potential Achilles heel for treatment. Here, we report the development and optimization of a fluorescence polarization-based competitive binding assay with LecA for application in screening of potential inhibitors. As a consequence of the low affinity of d-galactose for LecA, the fluorescent ligand was optimized to reduce protein consumption in the assay. The assay was validated using a set of known inhibitors of LecA and IC50 values in good agreement with the known Kd values were obtained. Finally, we employed the optimized assay to screen sets of synthetic thio-galactosides and natural blood group antigens and report their structure-activity relationship. In addition, we evaluated a multivalent fluorescent assay probe for LecA and report its applicability in an inhibition assay. PMID:27488655

  13. Lectin-like Oxidized Low-Density Lipoprotein (LDL) Receptor (LOX-1): A Chameleon Receptor for Oxidized LDL.

    Science.gov (United States)

    Zeya, Bushra; Arjuman, Albina; Chandra, Nimai Chand

    2016-08-16

    LOX-1, one of the main receptors for oxLDL, is found mainly on the surface of endothelial cells. It is a multifacet 52 kDa type II transmembrane protein that structurally belongs to the C-type lectin family. It exists with short intracellular N-terminal and long extracellular C-terminal hydrophilic domains separated by a hydrophobic domain of 26 amino acids. LOX-1 acts like a bifunctional receptor either showing pro-atherogenicity by activating the NFκB-mediated down signaling cascade for gene activation of pro-inflammatory molecules or playing an atheroprotective agent by receptor-mediated uptake of oxLDL in the presence of an anti-inflammatory molecule like IL-10. Mildly, moderately, and highly oxidized LDL show their characteristic features upon LOX-1 activation and its ligand binding indenture. The polymorphic LOX-1 genes are intensively associated with increased susceptibility to myocardial diseases. The splicing variant LOX IN dimerizes with the native form of LOX-1 and protects cells from damage by oxidized LDL. In the developing field of regenerating medicine, LOX-1 is a potential target for therapeutic intervention. PMID:27419271

  14. Carbohydrate specificity of lectin, purified from the fruiting bodies of Mycena pura /Fr./ Kumm. and its use in histochemical investigation

    Directory of Open Access Journals (Sweden)

    Ambarova N. O.

    2009-12-01

    Full Text Available Aim. The purpose of this investigation was to research carbohydrate specificity of a new lectin from fruiting body of Mycena pura and possibilities of its application in histochemical studies. Methods. The lectin has been purified by affinity chromatography on «îvomucine». The lectin carbohydrate specificity has been determined by a reaction of inhibiting haemagglutination by haptens. Histological materials were fixed in 4 % neutral formalin solution. Alkaline phosphatase was revealed in the cryostat unfixed microscopical sections. Results. The lectin yield from fresh fruit bodies of raw material was 9 mg/kg. Mol. mass of the lectin is 40 kDa. The lectin poorly interacted with D-glucose and D-mannose in contrast to lectins from Pisum sativum and Leucojum vernum. The peculiarity of this lectin is its strong interaction with alkaline phosphatase, the highest among twenty tested lectins. However, the receptors for Mycena lectin binding in mammalian tissues are not limited by this enzyme being presented also by glycoconjugates of another structure, as it was shown for fetus calf small intestine and kidney of rat. Conclusions. An important role in the lectin interaction with glycoproteins probably belongs to the disaccharide links of GlcNAcb(1-2Mana(1-6 or GlcNAcb(1- 2Mana(1-2, which not necessarily are terminal

  15. Processing-independent analysis for pro-C-type natriuretic peptide

    DEFF Research Database (Denmark)

    Lippert, Solvej Kølvraa; Rehfeld, Jens F.; Gøtze, Jens Peter

    C-type natriuretic peptide (CNP) is expressed in several human tissues. We designed a specific processing-independent assay for proCNP-derived products and quantitated the concentrations in human seminal plasma from normal and vasectomized men. Antibodies were raised against the N-terminus of human...... proCNP 11-27. Samples were incubated with trypsin prior to immunoassay, which allows for the measurement of "total" proCNP irrespective of the degree of post-translational processing. Seminal plasma from normal young men and vasectomized men were collected and quantitated; the molecular heterogeneity...... 104-933 pmol/L, age 18-25 years); gel chromatography suggested the presence of several molecular forms. Parameters associated to male fertility, proCNP concentrations in blood plasma and time of abstinence did not correlate to the seminal proCNP concentrations. Measurement in vasectomized men...

  16. Comparison of the Window-Frame RHIC-abort kicker with C-type Kicker

    Energy Technology Data Exchange (ETDEWEB)

    Tsoupas, N. [Brookhaven National Lab. (BNL), Upton, NY (United States). Collider-Accelerator Dept.; Hahn, H. [Brookhaven National Lab. (BNL), Upton, NY (United States). Collider-Accelerator Dept.; Meng, W. [Brookhaven National Lab. (BNL), Upton, NY (United States). Collider-Accelerator Dept.; Severance, Michael [Stony Brook Univ., NY (United States); McMahan, Brandon [Westhampton High School, NY (United States)

    2014-08-26

    The high intensity proton bunches (~2.5x1011 p/bunch ) circulating in RHIC increase the temperature of the ferrite-made RHIC-abort-kickers above the Curie point; as a result, the kickers cannot provide the required field to abort the beam at the beam dump. A team of experts in the CAD department worked on modifying the design of the window-frame RHIC-abort kicker to minimize the hysteresis losses responsible for the increase of the ferrite’s temperature. In this technical note we report some results from the study of two possible modifications of the window-frame RHIC-abort kicker, and we compare these results with those of a propose C-type RHIC-abort kicker. We also include an Appendix where we describe a method which may further reduce the hysteresis losses of the window-frame kicker.

  17. Comparison of the Window-Frame RHIC-abort kicker with C-type Kicker

    International Nuclear Information System (INIS)

    The high intensity proton bunches (~2.5x1011 p/bunch ) circulating in RHIC increase the temperature of the ferrite-made RHIC-abort-kickers above the Curie point; as a result, the kickers cannot provide the required field to abort the beam at the beam dump. A team of experts in the CAD department worked on modifying the design of the window-frame RHIC-abort kicker to minimize the hysteresis losses responsible for the increase of the ferrite's temperature. In this technical note we report some results from the study of two possible modifications of the window-frame RHIC-abort kicker, and we compare these results with those of a propose C-type RHIC-abort kicker. We also include an Appendix where we describe a method which may further reduce the hysteresis losses of the window-frame kicker.

  18. Crystallization and crystal manipulation of the Pterocarpus angolensis seed lectin.

    Science.gov (United States)

    Loris, Remy; Garcia-Pino, Abel; Buts, Lieven; Bouckaert, Julie; Beeckmans, Sonia; De Greve, Henri; Wyns, Lode

    2005-06-01

    The Man/Glc-specific legume lectin from the seeds of the African bloodwood tree (Pterocarpus angolensis) was crystallized in the presence of the disaccharide ligand Man(alpha1-3)ManMe. Small crystals initially appeared from a preliminary screen, but proved difficult to reproduce. The initial crystals were used to prepare microseeds, leading to a reproducible crystallization protocol. All attempts to obtain crystals directly of the ligand-free protein or of other carbohydrate complexes failed. However, the Man(alpha1-3)ManMe co-crystals withstand soaking with ten other carbohydrates known to bind to the lectin. Soaking for 15 min in 100 mM carbohydrate typically resulted in complete replacement of Man(alpha1-3)ManMe by the desired carbohydrate despite the involvement of lattice contacts at the binding site. Transferring the crystals for two weeks in carbohydrate-free artificial mother liquor resulted in the complete removal of the sugar from one of the two monomers in the asymmetric unit. Additional treatment of these crystals with 100 mM EDTA for two weeks resulted in removal of the structural calcium and manganese ions, which is accompanied by significant structural rearrangements of the loops that constitute the carbohydrate-binding site. PMID:15930620

  19. Magnetochrome: a c-type cytochrome domain specific to magnetotatic bacteria.

    Science.gov (United States)

    Siponen, Marina I; Adryanczyk, Géraldine; Ginet, Nicolas; Arnoux, Pascal; Pignol, David

    2012-12-01

    Magnetotactic bacteria consist of a group of taxonomically, physiologically and morphologically diverse prokaryotes, with the singular ability to align with geomagnetic field lines, a phenomenon referred to as magnetotaxis. This magnetotactic property is due to the presence of iron-rich crystals embedded in lipidic vesicles forming an organelle called the magnetosome. Magnetosomes are composed of single-magnetic-domain nanocrystals of magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4)) embedded in biological membranes, thereby forming a prokaryotic organelle. Four specific steps are described in this organelle formation: (i) membrane specialization, (ii) iron acquisition, (iii) magnetite (or greigite) biocrystallization, and (iv) magnetosome alignment. The formation of these magnetic crystals is a genetically controlled process, which is governed by enzyme-catalysed processes. On the basis of protein sequence analysis of genes known to be involved in magnetosome formation in Magnetospirillum magneticum AMB-1, we have identified a subset of three membrane-associated or periplasmic proteins containing a double cytochrome c signature motif CXXCH: MamE, MamP and MamT. The presence of these proteins suggests the existence of an electron-transport chain inside the magnetosome, contributing to the process of biocrystallization. We have performed heterologous expression in E. coli of the cytochrome c motif-containing domains of MamE, MamP and MamT. Initial biophysical characterization has confirmed that MamE, MamP and MamT are indeed c-type cytochromes. Furthermore, determination of redox potentials for this new family of c-type cytochromes reveals midpoint potentials of -76 and -32 mV for MamP and MamE respectively. PMID:23176475

  20. Recent advances in lectin-like oxidized low-density lipoprotein receptor-1 and atherosclerosis%血凝素样氧化低密度脂蛋白受体-1与动脉粥样硬化的研究进展

    Institute of Scientific and Technical Information of China (English)

    朱惠莲; 凌文华

    2004-01-01

    Lectin-like oxidized low density lipoprotein receptor-1 (LOX-1 } is a type-Ⅱ membrane protein belonging to the C-type lectin family molecules, which acts as a cell surface endocytosis receptor for atherogenic oxidized LDL (Ox-LDL). LOX-1 supports the binding internalization and proteolytic degradation of oxidized LDL, but not of significant amounts of acetylated LDL. LOX-1 is initially synthesized as a 40 kD precursor protein with N-linked high mannose-type carbohydrate, which is further glycosylated and processed into a 48-kD mature form. In vivo, endothelial cells that cover early therosclerotic lesions, intimal macrophages and smooth muscle cells in advanced atherosclerotic plaques express LOX-1. LOX-1 is cleaved at membrane proximal extracellular domain and released from the cell surface. Measurement of soluble LOX-1 in vivo may provide novel diagnostic strategy for the evaluation and prediction of atherosclerosis and vascular diseases.

  1. Structure of a lectin from Canavalia gladiata seeds: new structural insights for old molecules

    Directory of Open Access Journals (Sweden)

    Santi-Gadelha Tatiane

    2007-08-01

    Full Text Available Abstract Background Lectins are mainly described as simple carbohydrate-binding proteins. Previous studies have tried to identify other binding sites, which possible recognize plant hormones, secondary metabolites, and isolated amino acid residues. We report the crystal structure of a lectin isolated from Canavalia gladiata seeds (CGL, describing a new binding pocket, which may be related to pathogen resistance activity in ConA-like lectins; a site where a non-protein amino-acid, α-aminobutyric acid (Abu, is bound. Results The overall structure of native CGL and complexed with α-methyl-mannoside and Abu have been refined at 2.3 Å and 2.31 Å resolution, respectively. Analysis of the electron density maps of the CGL structure shows clearly the presence of Abu, which was confirmed by mass spectrometry. Conclusion The presence of Abu in a plant lectin structure strongly indicates the ability of lectins on carrying secondary metabolites. Comparison of the amino acids composing the site with other legume lectins revealed that this site is conserved, providing an evidence of the biological relevance of this site. This new action of lectins strengthens their role in defense mechanisms in plants.

  2. Structure of Dioclea virgata lectin: relations between carbohydrate binding site and nitric oxide production

    Energy Technology Data Exchange (ETDEWEB)

    Delatorre, P.; Gadelha, C.A.A.; Santi-Gadelha, T. [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Nobrega, R.B.; Rocha, B.A.M.; Nascimento, K.S.; Naganao, C.S.; Sampaio, A.H.; Cavada, B.S. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil); Pires, A.F.; Assreuy, A.M.S. [Universidade Estadual do Ceara (UECE), Fortaleza, CE (Brazil)

    2012-07-01

    Full text: Lectins are proteins/glycoproteins with at least one noncatalytic domain binding reversibly to specific monosaccharides or oligosaccharides. By binding to carbohydrate moieties on the cell surface, lectins participate in a range of cellular processes without changing the properties of the carbohydrates involved. The lectin of Dioclea virgata (DvirL), both native and complexed with X-man, was submitted to X-ray diffraction analysis and the crystal structure was compared to that of other Diocleinae lectins in order to better understand differences in biological proper- ties, especially with regard to the ability of lectins to induce nitric oxide (NO) production. The DvirL diffraction analysis revealed that both the native crystal and the X-Man-complexed form are orthorhombic and belong to space group I222. The cell parameters were: a=65.4 , b=86.6 and c=90.2 (native structure), and a=61.89 , b=87.67 and c=88.78 (X-Man-complexed structure). An association was observed between the volume of the carbohydrate recognition domain (CRD), the ability to induce NO production and the relative positions of Tyr12, Arg228 and Leu99. Thus, differences in biological activity induced by Diocleinae lectins are related to the configuration of amino acid residues in the carbohydrate binding site and to the structural conformation of subsequent regions capable of influencing site-ligand interactions. In conclusion, the ability of Diocleinae lectins to induce NO production depends on CRD configuration. (author)

  3. Lectin typing of Campylobacter jejuni using a novel quartz crystal microbalance technique

    Energy Technology Data Exchange (ETDEWEB)

    Yakovleva, Maria E., E-mail: maria.yakovleva@gmail.com [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Moran, Anthony P. [Department of Microbiology, School of Natural Sciences, National University of Ireland, Galway (Ireland); Safina, Gulnara R. [Department of Analytical and Marine Chemistry, University of Gothenburg, 412 96 Gothenburg (Sweden); Wadstroem, Torkel [Department of Infectious Diseases and Medical Microbiology, Lund University, 223 62 Lund (Sweden); Danielsson, Bengt [Acromed Invest AB, Magistratsvaegen 10, 226 43 Lund (Sweden)

    2011-05-23

    Seven Campylobacter jejuni strains were characterised by a lectin typing assay. The typing system was based on a quartz crystal microbalance technique (QCM) with four commercially available lectins (wheat germ agglutinin, Maackia amurensis lectin, Lens culinaris agglutinin, and Concanavalin A), which were chosen for their differing carbohydrate specificities. Initially, the gold surfaces of the quartz crystals were modified with 11-mercaptoundecanoic acid followed by lectin immobilisation using a conventional amine-coupling technique. Bacterial cells were applied for lectin typing without preliminary treatment, and resonant frequency and dissipation responses were recorded. The adhesion of microorganisms on lectin surfaces was confirmed by atomic force microscopy. Scanning was performed in the tapping mode and the presence of bacteria on lectin-coated surfaces was successfully demonstrated. A significant difference in the dissipation response was observed for different C. jejuni strains which made it possible to use this parameter for discriminating between bacterial strains. In summary, the QCM technique proved a powerful tool for the recognition and discrimination of C. jejuni strains. The approach may also prove applicable to strain discrimination of other bacterial species, particularly pathogens.

  4. Quantitation of two endogenous lactose-inhibitable lectins in embryonic and adult chicken tissues

    International Nuclear Information System (INIS)

    Two lactose-binding lectins from chicken tissues, chicken-lactose-lectin-I (CLL-I) and chicken-lactose-lectin-II (CLL-II) were quantified with a radioimmunoassay in extracts of a number of developing and adult chicken tissues. Both lectins could be measured in the same extract without separation, because they showed no significant immunological cross- reactivity. Many embryonic and adult tissues, including brain, heart, intestine, kidney, liver, lung, muscle, pancreas, and spleen, contained one or both lectins, although their concentrations differed markedly. For example, embryonic muscle, the richest source of CLL-I contained only traces of CLL-II whereas embryonic kidney, a very rich source of CLL-II contained substantial CLL-I. In both muscle and kidney, lectin levels in adulthood were much lower than in the embryonic state. In contrast, CLL-I in liver and CLL-II in intestine were 10-fold to 30-fold more concentrated in the adult than in the 15-d embryo. CLL-I and CLL-II from several tissues were purified by affinity chromatography and their identity in the various tissues was confirmed by polyacrylamide gel electrophoresis, isoelectric focusing, and peptide mapping. The results suggest that these lectins might have different functions in the many developing and adult tissues in which they are found

  5. The Lectin Frontier Database (LfDB, and Data Generation Based on Frontal Affinity Chromatography

    Directory of Open Access Journals (Sweden)

    Jun Hirabayashi

    2015-01-01

    Full Text Available Lectins are a large group of carbohydrate-binding proteins, having been shown to comprise at least 48 protein scaffolds or protein family entries. They occur ubiquitously in living organisms—from humans to microorganisms, including viruses—and while their functions are yet to be fully elucidated, their main underlying actions are thought to mediate cell-cell and cell-glycoconjugate interactions, which play important roles in an extensive range of biological processes. The basic feature of each lectin’s function resides in its specific sugar-binding properties. In this regard, it is beneficial for researchers to have access to fundamental information about the detailed oligosaccharide specificities of diverse lectins. In this review, the authors describe a publicly available lectin database named “Lectin frontier DataBase (LfDB”, which undertakes the continuous publication and updating of comprehensive data for lectin-standard oligosaccharide interactions in terms of dissociation constants (Kd’s. For Kd determination, an advanced system of frontal affinity chromatography (FAC is used, with which quantitative datasets of interactions between immobilized lectins and >100 fluorescently labeled standard glycans have been generated. The FAC system is unique in its clear principle, simple procedure and high sensitivity, with an increasing number (>67 of associated publications that attest to its reliability. Thus, LfDB, is expected to play an essential role in lectin research, not only in basic but also in applied fields of glycoscience.

  6. Isolation and analysis of mannose/trehalose/maltose specific lectin from jack bean with antibruchid activity.

    Science.gov (United States)

    Shanmugavel, Sakthivelkumar; Velayutham, Veeramani; Kamalanathan, Tamilarasan; Periasamy, Mullainadhan; Munusamy, Arumugam; Sundaram, Janarthanan

    2016-10-01

    A lectin with insecticidal property against the stored product pest, Callosobruchus maculatus was successfully isolated from the seeds of Canavalia virosa using standard affinity chromatography. The isolated molecule typically behaved like a lectin in its characteristics. It agglutinated indicator red blood cells (RBC) in its native as well as enzyme treated conditions. The enzyme treated RBC types exhibited a very high hemagglutination (HA) titre values and this property of isolated molecule behaved like arcelin, the lectin-like molecules reported from several species of Phaseolus. As a characteristic feature of a lectin, the isolated molecule effectively inhibited the agglutination of indicator RBC types with simple and complex carbohydrates including glycoproteins. This nature of the isolated molecule also relate with characteristic feature of arcelin isoforms in inhibiting HA activity with complex glycoproteins as reported in many studies. Most interestingly, the present study disclosed trehalose as a potent inhibitor of C. virosa lectin. Therefore, feeding insect pests on the lectin like arcelin could serve as antibiosis factor/anti-insect activity. The molecular characteristics of this isolated molecule and its mass studies too revealed its homology with arcelin, arcelin-1, 2 and 6 isoforms of P. vulgaris and lectin from Canavalia cathartica, C. lineata and C. brasiliensis. PMID:27238584

  7. Isolation and characterization of a new mannose-binding lectin gene from Taxus media

    Indian Academy of Sciences (India)

    Guoyin Kai; Lingxia Zhao; Jingui Zheng; Lei Zhang; Zhiqi Miao; Xiaofen Sun; Kexuan Tanga

    2004-12-01

    In this paper, we report the cloning and characterization of the first mannose-binding lectin gene from a gymnosperm plant species, Taxus media. The full-length cDNA of T. media agglutinin (TMA) consisted of 676 bp and contained a 432 bp open reading frame (ORF) encoding a 144 amino acid protein. Comparative analysis showed that TMA had high homology with many previously reported plant mannose-binding lectins and that tma encoded a precursor lectin with a 26-aa signal peptide. Molecular modelling revealed that TMA was a new mannose-binding lectin with three typical mannose-binding boxes like lectins from species of angiosperms. Tissue expression pattern analyses revealed that tma is expressed in a tissue-specific manner in leaves and stems, but not in fruits and roots. Phylogenetic tree analyses showed that TMA belonged to the structurally and evolutionarily closely related monocot mannose-binding lectin superfamily. This study provides useful information to understand the molecular evolution of plant lectins.

  8. The effect of lectin from Taro tuber (Colocasia antiquorum) given by force-feeding on the growth of mice.

    Science.gov (United States)

    Seo, Y J; Une, S; Tsukamoto, I; Miyoshi, M

    1990-06-01

    In earlier experiments in our laboratory, a lectin from the Kintoki bean (Phaseolus vulgaris) was found to have not only erythrocyte agglutinating activity but also toxicities for mice and rats, including growth inhibitory activity and even lethal activity. A number of studies on legume lectins have been carried out in other laboratories as well. But relatively little attention has been paid to lectins from non-leguminous foods. In the present study, we chose Taro tuber as a source of non-leguminous lectins and prepared two types of Taro tuber lectin. One was crude lectin precipitated with ammonium sulfate from the aqueous extract and the other was pure lectin isolated as we described previously. The two were compared with regards to the antinutritional functions in mice. The daily doses were 100 mg for either intact or autoclaved crude lectin, which was a maximum amount available to give to mice in 1 ml, and 30 mg for the pure lectin which was equivalent to 100 mg of the crude lectin in hemagglutinating activity. Control mice were given 1 ml of water and the experiment was conducted for 6 days. Growth retardation was found in the mice given either lectin, but no significant difference was found in the weight increase between the control group and the autoclaved lectin group. For 3 days during the experimental period, physical activity was measured as an index of vigor of mice. The activities of the crude lectin and the pure lectin groups leveled down to 62.9 and 64.2% of that of the control group, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2292730

  9. Studies of hemolytical and antimicrobical action of Amanita virosa Secr. and Mycena pura /Fr./ Kumm. poisonous mushrooms lectins

    Directory of Open Access Journals (Sweden)

    Danileuchenko V. V.

    2010-02-01

    Full Text Available Aim. To study hemolytical and antimicrobical action of two new lectins, obtained from fruit bodies of poisonous basidial mushrooms of A. virosa Secr. and M. pura /Fr./ Kumm. Methods. Research on hemolytical action of lectins was conducted on the erythrocytes of human and animals. The experiments on osmotic protection of erythrocytes were performed in the presence of polyethylenglycols of different molecular mass (in a range from 400 to 4000 Da. Antimicrobical activity of lectins was studied by determination of area delay of growth of culture of different types of microorganisms on the Petri dish in an agaric media. Results. Both lectins hemolyse the erythrocytes of rabbit, human, rat and dog and do not hemolyse the erythrocytes of cow and ship in concentration of 1 mg/ml. The rabbit erythrocytes are most sensitive to hemolytical action of lectins, while hemolytic ability of A. virosa lectin is higher. Hemolysis was not observed in the presence of PEG of molecular mass over 1,350 Da. Action of lectins on 10 types of microorganisms was investigated. Lectins inhibited mainly growth of grammpositive microorganisms and protey. For most tested microorganisms antimicrobial action of Mycena lectin is stronger comparing with A. virosa lectin. Conclusions. Two new hemolytical lectins are found in the fruit bodies of mushrooms-basidiomycetes. The lectin formed ion-permeable pores in membrane of erythrocytes with the hydrodynamic diameter smaller than 2.3 nm and larger than 1.6 nm. These lectins displays also antimicrobial activity and by the sum of these features are similar to the cytolytic lectins of lower invertebrates.

  10. Evolutionary history and stress regulation of the lectin superfamily in higher plants

    Directory of Open Access Journals (Sweden)

    Ramachandran Srinivasan

    2010-03-01

    Full Text Available Abstract Background Lectins are a class of carbohydrate-binding proteins. They play roles in various biological processes. However, little is known about their evolutionary history and their functions in plant stress regulation. The availability of full genome sequences from various plant species makes it possible to perform a whole-genome exploration for further understanding their biological functions. Results Higher plant genomes encode large numbers of lectin proteins. Based on their domain structures and phylogenetic analyses, a new classification system has been proposed. In this system, 12 different families have been classified and four of them consist of recently identified plant lectin members. Further analyses show that some of lectin families exhibit species-specific expansion and rapid birth-and-death evolution. Tandem and segmental duplications have been regarded as the major mechanisms to drive lectin expansion although retrogenes also significantly contributed to the birth of new lectin genes in soybean and rice. Evidence shows that lectin genes have been involved in biotic/abiotic stress regulations and tandem/segmental duplications may be regarded as drivers for plants to adapt various environmental stresses through duplication followed by expression divergence. Each member of this gene superfamily may play specialized roles in a specific stress condition and function as a regulator of various environmental factors such as cold, drought and high salinity as well as biotic stresses. Conclusions Our studies provide a new outline of the plant lectin gene superfamily and advance the understanding of plant lectin genes in lineage-specific expansion and their functions in biotic/abiotic stress-related developmental processes.

  11. Differential activity of a lectin from Solieria filiformis against human pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    M.L. Holanda

    2005-12-01

    Full Text Available A lectin isolated from the red alga Solieria filiformis was evaluated for its effect on the growth of 8 gram-negative and 3 gram-positive bacteria cultivated in liquid medium (three independent experiments/bacterium. The lectin (500 µg/mL stimulated the growth of the gram-positive species Bacillus cereus and inhibited the growth of the gram-negative species Serratia marcescens, Salmonella typhi, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus sp, and Pseudomonas aeruginosa at 1000 µg/mL but the lectin (10-1000 µg/mL had no effect on the growth of the gram-positive bacteria Staphylococcus aureus and B. subtilis, or on the gram-negative bacteria Escherichia coli and Salmonella typhimurium. The purified lectin significantly reduced the cell density of gram-negative bacteria, although no changes in growth phases (log, exponential and of decline were observed. It is possible that the interaction of S. filiformis lectin with the cell surface receptors of gram-negative bacteria promotes alterations in the flow of nutrients, which would explain the bacteriostatic effect. Growth stimulation of the gram-positive bacterium B. cereus was more marked in the presence of the lectin at a concentration of 1000 µg/mL. The stimulation of the growth of B. cereus was not observed when the lectin was previously incubated with mannan (125 µg/mL, its hapten. Thus, we suggest the involvement of the binding site of the lectin in this effect. The present study reports the first data on the inhibition and stimulation of pathogenic bacterial cells by marine alga lectins.

  12. New perspectives on mannan-binding lectin-mediated complement activation

    DEFF Research Database (Denmark)

    Degn, Søren Egedal; Thiel, Steffen; Jensenius, Jens Christian

    2007-01-01

    The complement system is an important part of the innate immune system, mediating several major effector functions and modulating adaptive immune responses. Three complement activation pathways exist: the classical pathway (CP), the alternative pathway (AP), and the lectin pathway (LP). The LP is......, allowing C3 activation in the absence of components otherwise believed critical. The classical bypass pathways are dependent on C1 and components of the AP. A recent study has shown the existence also of a lectin bypass pathway dependent on mannan-binding lectin (MBL) and AP components. The emerging...

  13. Purification of a lectin from Eugenia uniflora L. seeds and its potential antibacterial activity

    OpenAIRE

    Oliveira, M. D. L.; Andrade, C. A. S.; Magalhães, N. S.; L.C.B.B. Coelho; Teixeira, J.A.; Cunha, M. G. C.; Correia, M. T. S.

    2008-01-01

    Aims: The aim of this work was to analyse the antimicrobial properties of a purified lectin from Eugenia uniflora L. seeds. Methods and Results: The E. uniflora lectin (EuniSL) was isolated from the seed extract and purified by ion-exchange chromatography in DEAE-Sephadex with a purification factor of 11·68. The purified lectin showed a single band on denaturing electrophoresis, with a molecular mass of 67 kDa. EuniSL agglutinated rabbit and human erythrocytes with a higher specificity fo...

  14. Molecular characterization of a c-type lysozyme from the desert locust, Schistocerca gregaria (Orthoptera: Acrididae).

    Science.gov (United States)

    Mohamed, Amr A; Zhang, Long; Dorrah, Moataza A; Elmogy, Mohamed; Yousef, Hesham A; Bassal, Taha T M; Duvic, Bernard

    2016-08-01

    Lysozymes are bacteriolytic peptides that are implicated in the insect nonspecific innate immune responses. In this study, a full-length cDNA encoding a c-type lysozyme from Schistocerca gregaria (SgLys) has been cloned and characterized from the fat body of immune-challenged 5(th) instar. The deduced mature lysozyme is 119 amino acid residues in length, has a calculated molecular mass of 13.4 kDa and an isoelectric point (Ip) of 9.2. SgLys showed high identities with other insect lysozymes, ranging from 41.5% to 93.3% by BLASTp search in NCBI. Eukaryotic in vitro expression of the SgLys ORF (rSgLys) with an apparent molecular mass of ∼16 kDa under SDS-PAGE is close to the calculated molecular weight of the full-length protein. rSgLys displayed growth inhibitory activity against Gram-negative and Gram-positive bacteria. 3D structure modeling of SgLys, based on comparison with that of silkworm lysozyme, and sequence comparison with the helix-loop-helix (α-hairpin) structure of hen egg white lysozyme (HEWL) were employed to interpret the antibacterial potencies. Phylogenetic alignments indicate that SgLys aligns well with insect c-type lysozymes that expressed principally in fat body and hemocytes and whose role has been defined as immune-related. Western blot analysis showed that SgLys expression was highest at 6-12 h post-bacterial challenge and subsequently decreased with time. Transcriptional profiles of SgLys were determined by semi-quantitative RT-PCR analysis. SgLys transcript was upregulated at the highest level in fat body, hemocytes, salivary gland, thoracic muscles, and epidermal tissue. It was expressed in all developmental stages from egg to adult. These data indicate that SgLys is a predominant acute-phase protein that is expressed and upregulated upon immune challenge. PMID:26997372

  15. B and C types natriuretic peptides modify norepinephrine uptake and release in the rat adrenal medulla.

    Science.gov (United States)

    Vatta, M S; Presas, M F; Bianciotti, L G; Rodriguez-Fermepin, M; Ambros, R; Fernandez, B E

    1997-01-01

    We have previously reported that atrial natriuretic factor (ANF) modulates adrenomedullar norepinephrine (NE) metabolism. On this basis, the aim of the present work was to study the effects of B and C types natriuretic peptides (BNP and CNP) on the uptake, intracellular distribution and release of 3H-NE. Experiments were carried out in rat adrenal medulla slices incubated "in vitro." Results showed that 100 nM of both, CNP and BNP, enhanced total and neuronal NE uptake. Both peptides (100 nM) caused a rapid increase in NE uptake during the first minute, which was sustained for 60 min. NE intracellular distribution was only modified by CNP (100 nM), which increased the granular fraction and decreased the cytosolic pool. On the other hand, spontaneous as well as evoked (KCl) NE release, was decreased by BNP and CNP (50 and 100 nM for spontaneous release and 1, 10, 50 and 100 nM for evoked output). The present results suggest that BNP and CNP may regulate catecholamine secretion and modulate adrenomedullary biological actions mediated by catecholamines, such as blood arterial pressure, smooth muscle tone, and metabolic activities. PMID:9437706

  16. Thickness controlled proximity effects in C-type antiferromagnet/superconductor heterostructure.

    Science.gov (United States)

    Mani, Awadhesh; Kumary, T Geetha; Lin, J G

    2015-01-01

    Modulation of the superconducting state possessing a C-type antiferromagnetic phase in the Nd0.35Sr0.65MnO3/YBa2Cu3O7 heterostructure is investigated, with the Nd0.35Sr0.65MnO3 thickness (t) varying from 40 to 200 nm. Both the superconducting transition temperature and the upper critical field along the c-axis decrease with increasing t; while the in-plane coherence length increases from 2.0 up to 3.6 nm. Meanwhile, the critical current density exhibits a field-independent behavior, indicating an enhanced flux pinning effect. Furthermore, low-temperature spin canting induces a breakdown and re-entrance of the superconductivity, demonstrating a dynamic completion between the superconducting pairing and the exchange field. An unexpected colossal magnetoresistance is observed below the superconducting re-entrance temperature at t = 200 nm, which is attributed to the dominant influence of the exchange field over the pairing energy. PMID:26239479

  17. Nutritional significance of lectins and enzyme inhibitors from legumes.

    Science.gov (United States)

    Lajolo, Franco M; Genovese, Maria Inés

    2002-10-23

    Legumes have natural components, such as lectins, amylase, and trypsin inhibitors, that may adversely affect their nutritional properties. Much information has already been obtained on their antinutritional significance and how to inactivate them by proper processing. Chronic ingestion of residual levels is unlikely to pose risks to human health. On the other hand, the ability of these molecules to inhibit some enzymes such as trypsin, chymotrypsin, disaccharidases, and alpha-amylases, to selectively bind to glycoconjugates, and to enter the circulatory system may be a useful tool in nutrition and pharmacology. Trypsin inhibitors have also been studied as cancer risk reducing factors. These components seem to act as plant defense substances. However, increased contents may represent an impairment of the nutritional quality of legumes because these glycoproteins and the sulfur-rich protease inhibitors have been shown to be poorly digested and to participate in chemical reactions during processing reducing protein digestibility, a still unsolved question. PMID:12381157

  18. Detection of Sugar-Lectin Interactions by Multivalent Dendritic Sugar Functionalized Single-Walled Carbon Nanotubes

    CERN Document Server

    Vasu, K S; Bagul, R S; Jayaraman, N; Sood, A K; 10.1063/1.4739793

    2012-01-01

    We show that single walled carbon nanotubes (SWNT) decorated with sugar functionalized poly (propyl ether imine) (PETIM) dendrimer is a very sensitive platform to quantitatively detect carbohydrate recognizing proteins, namely, lectins. The changes in electrical conductivity of SWNT in field effect transistor device due to carbohydrate - protein interactions form the basis of present study. The mannose sugar attached PETIM dendrimers undergo charge - transfer interactions with the SWNT. The changes in the conductance of the dendritic sugar functionalized SWNT after addition of lectins in varying concentrations were found to follow the Langmuir type isotherm, giving the concanavalin A (Con A) - mannose affinity constant to be 8.5 x 106 M-1. The increase in the device conductance observed after adding 10 nM of Con A is same as after adding 20 \\muM of a non - specific lectin peanut agglutinin, showing the high specificity of the Con A - mannose interactions. The specificity of sugar-lectin interactions was chara...

  19. Mannan binding lectin : a two-faced regulator of renal allograft injury?

    NARCIS (Netherlands)

    Damman, Jeffrey; Seelen, Marc A.

    2013-01-01

    Complement activation plays an important role in the pathogenesis of renal allograft injury after kidney transplantation. There are three known pathways of complement activation, namely, classical, alternative, and lectin pathways. In renal allograft injury, contradictory results were reported about

  20. Change in cell wall lectin activity and some phytohormones in tobacco leaves during photoperiodic induction

    International Nuclear Information System (INIS)

    The change in cell wall lectin activity and some phytohormones in the Silvestris and Mamont tobacco leaves during photoperiodic induction is studied. The ratio between two types of tobaccos changes alike

  1. Superparamagnetic adsorbents for high-gradient magnetic fishing of lectins out of legume extracts

    DEFF Research Database (Denmark)

    Heebøll-Nielsen, Anders; Dalkiær, M.; Hubbuch, Jürgen;

    2004-01-01

    This work presents the development, testing, and application in high-gradient magnetic fishing of superparamagnetic supports for adsorption of lectins. Various approaches were examined to produce affinity, mixed mode, and hydrophobic charge induction type adsorbents. In clean monocomponent system...

  2. Reactivities of N-acetylgalactosamine-specific lectins with human IgA1 proteins

    DEFF Research Database (Denmark)

    Moore, J.S.; Kulhavy, R.; Tomana, M.;

    2007-01-01

    Lectins are proteins with specificity of binding to certain monosaccharides or oligosaccharides. They can detect abnormal glycosylation patterns on immunoglobulins in patients with various chronic inflammatory diseases, including rheumatoid arthritis and IgA nephropathy (IgAN). However, lectins...... exhibit binding heterogeneity, depending on their source and methods of isolation. To characterize potential differences in recognition of terminal N-acetylgalactosamine (GalNAc) on IgA1, we evaluated the binding characteristics of several commercial preparations of GalNAc-specific lectins using a panel...... of IgA1 and, as controls, IgA2 and IgG myeloma proteins. These lectins originated from snails Helix aspersa (HAA) and Helix pomatia (HPA), and the plant Vicia villosa (VV). Only HAA and HPA bound exclusively to IgA1, with its O-linked glycans composed of GalNAc, galactose, and sialic acid. In...

  3. STUDY OF AZOSPIRILLUM LECTINS INFLUENCE ON HYDROGEN PEROXIDE PRODUCTION IN WHEAT-ROOTS

    Directory of Open Access Journals (Sweden)

    Alen’kina S.A.

    2009-12-01

    Full Text Available It was found that two cell-surface lectins isolated from the nitrogen-fixing soil bacterium Azospirillum brasilense Sp7 and from its mutant defective in lectin activity, A. brasilense Sp7.2.3 can stimulate rapid formation of hydrogen peroxide, associated with an increase in the activities of oxalate oxidase and peroxidase in the roots of wheat seedlings. The most advantageous and most rapidly induced pathway of hydrogen peroxide formation was the oxidation of oxalic acid by oxalate oxidase because in this case, a 10-min treatment of the roots with the lectins at 10 µg ml-1 was sufficient. The data from this study attest that the Azospirillum lectins can act as inducers of adaptation processes in the roots of wheat seedlings.

  4. Lectin Histochemical Study of Vasculogenesis During Rat Pituitary Morphogenesis

    Directory of Open Access Journals (Sweden)

    Ali Reza Ebrahimzadeh Bideskan

    2011-01-01

    Full Text Available Objective(s The aim of this study was to investigate glycoconjugates distribution patterns as well as their changes during the course of pituitary portal vasculogenesis and angiogenesis.Materials and MethodsFormalin fixed paraffin sections of 10 to 20 days of Sprague Dawly rat fetuses were processed for histochemical studies using four different horseradish peroxidase (HRP conjugated lectins. Orange peel fungus (OFA, Vicica villosa (VVA, Glycine max (SBA and Wistaria floribunda (WFA specific for α-L-Fucose, D-Gal, α, ß-D-GalNAc and D- GalNAc terminal sugars of glycoconjugates respectively.ResultsOur finding indicated that adenohypophysal cells reacted with OFA on gestational day 10 (E10 and increased progressively to E14. Staining intensity did not change from days 14 to17, then after increased following days to E20 significantly (P< 0.05. A few cells around Rathke’s pouch reacted with VVA on E13, increased to E14 and decreased significantly afterward (P< 0.05. Reaction of some cells around Rathke’s pouch reacted with SBA on E14. This visible reaction was the same as E18 and decreased later (P< 0.05. Many cells around Rathke’s pouch reacted with WFA on E13 and increased on E 14 and E15 and decreased thereafter (P< 0.05.ConclusionReactions of OFA and other tested lectins with endothelial cells around Rathke’s pouch and developing pars distalis were different. These results suggest that embryonic origin of hypophiseal pituitary portal (HPP system endothelial cells are not the same and our finding also indicated that glycoconjugates with terminal sugars α-L-Fucose, D-Gal, α, ß-D-GalNAc may play critical role(s in cell interactions and tissue differentiations such as vasculogensis and angiogenesis as well as other developmental precursors in formation of the pituitary gland.

  5. Interaction indole-3-acetic acid IAA with lectin Canavalia maritima seeds reveal new function of lectins in plant physiology

    Energy Technology Data Exchange (ETDEWEB)

    Silva Filho, J.C.; Santi-Gadelha, T.; Gadelha, C.A.A.; Delatorre, P. [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Teixeira, C.S.; Rocha, B.A.M.; Nobrega, R.B.; Alencar, K.L.L.; Cavada, B.S. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil)

    2012-07-01

    Full text: Lectins are a class of proteins of non-immune origin characterized by its capability in interacts specifically and reversibly to mono and oligosaccharides. In plant several possible roles have been suggested including their function in seed maturation, cell wall assembly, defense mechanisms, or rhizobial nodulation of legume roots. Nearly all application and proposed of the plant lectins are based on their specific carbohydrate binding. However, it has been reported that lectins from legumes, might interact with other molecules, such as non proteic amino acids and hydrophobic compounds. This study show the first the crystal structure based on molecular replacement of the Canavalia maritima (CML) complexed with IAA correlated with possible role in plant development. Purified CML was dissolved in 20 mMTrisHCl pH 7.6 containing 5 mM IAA, the suitable co-crystals from CML-IAA complex grew in condition 4 of screen I (0.1 M TrisHCl pH 8.5 and 2.0 M ammonium sulfate). This crystal belong to the orthorhombic space group I222 with unit-cell parameters a = 67.1 ; b = 70.7 , c = 97.7 , The structure was refined at 2.1 of resolution to a final R factor of 20.63 % and an R free of 22.54 %. To check the relative position of the IAA molecule in relation to the biological assemble of the CML, the tetrameric structure was generate by crystallographic symmetry. IAA molecules are positioned in the central cavity. The IAA is stabilized by interacting through hydrogen bounds and Van der Waals forces with the amino acids residues Ser 108 and Asn131, and two water molecules. The hydrophilic interactions occur between IAA and side chains of Ser 108, Asn131 and water molecules 26 and 31 by H-bonds. The OG oxygen from Ser108 display H-bonds with O2 and O3 oxygen atoms from IAA, 3.1 and 2.8 respectively. The tetrameric structure of CML complexed with IAA revels which this protein can act during the seedling in plant development. (author)

  6. Interaction indole-3-acetic acid IAA with lectin Canavalia maritima seeds reveal new function of lectins in plant physiology

    International Nuclear Information System (INIS)

    Full text: Lectins are a class of proteins of non-immune origin characterized by its capability in interacts specifically and reversibly to mono and oligosaccharides. In plant several possible roles have been suggested including their function in seed maturation, cell wall assembly, defense mechanisms, or rhizobial nodulation of legume roots. Nearly all application and proposed of the plant lectins are based on their specific carbohydrate binding. However, it has been reported that lectins from legumes, might interact with other molecules, such as non proteic amino acids and hydrophobic compounds. This study show the first the crystal structure based on molecular replacement of the Canavalia maritima (CML) complexed with IAA correlated with possible role in plant development. Purified CML was dissolved in 20 mMTrisHCl pH 7.6 containing 5 mM IAA, the suitable co-crystals from CML-IAA complex grew in condition 4 of screen I (0.1 M TrisHCl pH 8.5 and 2.0 M ammonium sulfate). This crystal belong to the orthorhombic space group I222 with unit-cell parameters a = 67.1 ; b = 70.7 , c = 97.7 , The structure was refined at 2.1 of resolution to a final R factor of 20.63 % and an R free of 22.54 %. To check the relative position of the IAA molecule in relation to the biological assemble of the CML, the tetrameric structure was generate by crystallographic symmetry. IAA molecules are positioned in the central cavity. The IAA is stabilized by interacting through hydrogen bounds and Van der Waals forces with the amino acids residues Ser 108 and Asn131, and two water molecules. The hydrophilic interactions occur between IAA and side chains of Ser 108, Asn131 and water molecules 26 and 31 by H-bonds. The OG oxygen from Ser108 display H-bonds with O2 and O3 oxygen atoms from IAA, 3.1 and 2.8 respectively. The tetrameric structure of CML complexed with IAA revels which this protein can act during the seedling in plant development. (author)

  7. Differential effect of plant lectins on mast cells of different origins

    Directory of Open Access Journals (Sweden)

    F.C. Lopes

    2005-06-01

    Full Text Available Histamine release induced by plant lectins was studied with emphasis on the carbohydrate specificity, external calcium requirement, metal binding sites, and mast cell heterogeneity and on the importance of antibodies bound to the mast cell membrane to the lectin effect. Peritoneal mast cells were obtained by direct lavage of the rat peritoneal cavity and guinea pig intestine and hamster cheek pouch mast cells were obtained by dispersion with collagenase type IA. Histamine release was induced with concanavalin A (Con A, lectins from Canavalia brasiliensis, mannose-specific Cymbosema roseum, Maackia amurensis, Parkia platycephala, Triticum vulgaris (WGA, and demetallized Con A and C. brasiliensis, using 1-300 µg/ml lectin concentrations applied to Wistar rat peritoneal mast cells, peaking on 26.9, 21.0, 29.1, 24.9, 17.2, 10.7, 19.9, and 41.5%, respectively. This effect was inhibited in the absence of extracellular calcium. The lectins were also active on hamster cheek pouch mast cells (except demetallized Con A and on Rowett nude rat (animal free of immunoglobulins peritoneal mast cells (except for mannose-specific C. roseum, P. platycephala and WGA. No effect was observed in guinea pig intestine mast cells. Glucose-saturated Con A and C. brasiliensis also released histamine from Wistar rat peritoneal mast cells. These results suggest that histamine release induced by lectins is influenced by the heterogeneity of mast cells and depends on extracellular calcium. The results also suggest that this histamine release might occur by alternative mechanisms, because the usual mechanism of lectins is related to their binding properties to metals from which depend the binding to sugars, which would be their sites to bind to immunoglobulins. In the present study, we show that the histamine release by lectins was also induced by demetallized lectins and by sugar-saturated lectins (which would avoid their binding to other sugars. Additionally, the lectins

  8. Stability of Curcuma longa rhizome lectin: Role of N-linked glycosylation.

    Science.gov (United States)

    Biswas, Himadri; Chattopadhyaya, Rajagopal

    2016-04-01

    Curcuma longa rhizome lectin, a mannose-binding protein of non-seed portions of turmeric, is known to have antifungal, antibacterial and α-glucosidase inhibitory activities. We studied the role of complex-type glycans attached to asparagine (Asn) 66 and Asn 110 to elucidate the role of carbohydrates in lectin activity and stability. Apart from the native lectin, the characteristics of a deglycosylated Escherichia coli expressed lectin, high-mannose oligosaccharides at both asparagines and its glycosylation mutants N66Q and N110Q expressed in Pichia pastoris, were compared to understand the relationship between glycosylation and activity. Far UV circular dichroism (CD) spectra, fluorescence emission maximum, hemagglutination assay show no change in secondary or tertiary structures or sugar-binding properties between wild-type and aforementioned recombinant lectins under physiological pH. But reduced agglutination activity and loss of tertiary structure are observed in the acidic pH range for the deglycosylated and the N110Q protein. In thermal and guanidine hydrochloride (GdnCl)-induced unfolding, the wild-type and high-mannose lectins possess higher stability compared with the deglycosylated recombinant lectin and both mutants, as measured by a higher Tm of denaturation or a greater free energy change, respectively. Reversibility experiments after thermal denaturation reveal that deglycosylated proteins tend to aggregate during thermal inactivation but the wild type shows a much greater recovery to the native state upon refolding. These results suggest that N-glycosylation in turmeric lectin is important for the maintenance of its proper folding upon changes in pH, and that the oligosaccharides help in maintaining the active conformation and prevent aggregation in unfolded or partially folded molecules. PMID:26603318

  9. Evolutionary history and stress regulation of the lectin superfamily in higher plants

    OpenAIRE

    Ramachandran Srinivasan; Ma Zhigang; Jiang Shu-Ye

    2010-01-01

    Abstract Background Lectins are a class of carbohydrate-binding proteins. They play roles in various biological processes. However, little is known about their evolutionary history and their functions in plant stress regulation. The availability of full genome sequences from various plant species makes it possible to perform a whole-genome exploration for further understanding their biological functions. Results Higher plant genomes encode large numbers of lectin proteins. Based on their doma...

  10. Glycophenotype Evaluation in Cutaneous Tumors Using Lectins Labeled with Acridinium Ester

    OpenAIRE

    Luiza Rayanna Amorim Lima; Matheus Filgueira Bezerra; Sinara Mônica Vitalino de Almeida; Lúcia Patrícia Bezerra Gomes Silva; Eduardo Isidoro Carneiro Beltrão; Luiz Bezerra de Carvalho Júnior

    2013-01-01

    Background. Tumor cells show alterations in their glycosylation patterns when compared to normal cells. Lectins can be used to evaluate these glycocode changes. Chemiluminescence assay is an effective technique for quantitative analysis of proteins, nucleic acids, and carbohydrates due to its high sensitivity, specificity, and rapid testing. Objective. To use histochemiluminescence based on lectin conjugated to acridinium ester (AE) for the investigation of glycophenotype changes in cutaneous...

  11. Lectin characterization of gonococci from an outbreak caused by penicillin-resistant Neisseria gonorrhoeae.

    OpenAIRE

    Schalla, W O; Rice, R J; Biddle, J W; Jeanlouis, Y; Larsen, S A; Whittington, W L

    1985-01-01

    A total of 40 Neisseria gonorrhoeae isolates, representing 19 penicillin-resistant isolates (from 8 heterosexual patients and 11 homosexual patients) and 21 penicillin-susceptible isolates (from 15 heterosexual patients and 6 homosexual patients) and obtained from the same geographic area, were examined. Lectin agglutination patterns were based on the reactivity of the isolates with the following 14 lectins: concanavalin A, Lens culinaris, Trichosanthes kinlowii, Griffonia simplicifolia I, Ar...

  12. Differentiation of Helicobacter pylori isolates based on lectin binding of cell extracts in an agglutination assay

    OpenAIRE

    Hynes, Sean; Hirmo, Siiri; Wadström, Torkel; Moran, Anthony P.

    1999-01-01

    Plant and animal lectins with various carbohydrate specificities were used to type 35 Irish clinical isolates of Helicobacter pylori and the type strain NCTC 11637 in a microtiter plate assay. Initially, a panel of eight lectins with the indicated primary specificities were used: Anguilla anguilla (AAA), Lotus tetragonolobus (Lotus A), and Ulex europaeus I (UEA I), specific for α-l-fucose; Solanum tuberosum (STA) and Triticum vulgaris (WGA), specific for β-N-acetylglucosamine; Glycine max (SB...

  13. Protective effect of lectin from Synadenium carinatum on Leishmania amazonensis infection in BALB/c mice

    OpenAIRE

    Afonso-Cardoso, Sandra R.; Rodrigues, Flávio H.; Gomes, Márcio A.B.; Silva, Adriano G.; Ademir ROCHA; Guimarães, Aparecida H.B.; Candeloro, Ignês; Favoreto, Sílvio; Marcelo S. Ferreira; de Souza, Maria A

    2007-01-01

    The protective effect of the Synadenium carinatum latex lectin (ScLL), and the possibility of using it as an adjuvant in murine model of vaccination against American cutaneous leishmaniasis, were evaluated. BALB/c mice were immunized with the lectin ScLL (10, 50, 100 µg/animal) separately or in association with the soluble Leishmania amazonensis antigen (SLA). After a challenge infection with 106 promastigotes, the injury progression was monitored weekly by measuring the footpad swelling for ...

  14. Reactivities of N-acetylgalactosamine-specific lectins with human IgA1 proteins

    OpenAIRE

    Moore, Jennifer S.; Kulhavy, Rose; Tomana, Milan; Moldoveanu, Zina; Brown, Rhubell; Hall, Stacy; Kilian, Mogens; Poulsen, Knud; Mestecky, Jiri; Julian, Bruce A.; Novak, Jan

    2007-01-01

    Lectins are proteins with specificity of binding to certain monosaccharides or oligosaccharides. They can detect abnormal glycosylation patterns on immunoglobulins in patients with various chronic inflammatory diseases, including rheumatoid arthritis and IgA nephropathy (IgAN). However, lectins exhibit binding heterogeneity, depending on their source and methods of isolation. To characterize potential differences in recognition of terminal N-acetylgalactosamine (GalNAc) on IgA1, we evaluated ...

  15. The Contribution of Mannose Binding Lectin to Reperfusion Injury after Ischemic Stroke

    OpenAIRE

    Morrison, Helena; Frye, Jennifer; Davis-Gorman, Grace; Funk, Janet; McDonagh, Paul; Stahl, Gregory; Ritter, Leslie

    2011-01-01

    After complement system (CS) activation, the sequential production of complement products increases cell injury and death through opsonophagocytosis, cytolysis, adaptive, and inflammatory cell responses. These responses potentiate cerebral ischemia-reperfusion (IR) injury after ischemic stroke and reperfusion. Activation of the CS via mannose binding lectin (MBL)-initiated lectin pathway is known to increase tissue damage in response to IR in muscle, myocardium and intestine tissue. In contra...

  16. Burn Injury Reveals Altered Phenotype in Mannan-Binding Lectin-Deficient Mice

    OpenAIRE

    Møller-Kristensen, Mette; Hamblin, Michael R.; Thiel, Steffen; Jensenius, Jens Chr.; Takahashi, Kazue

    2007-01-01

    Burn injury destroys skin, the second largest innate immune organ in the body, and triggers chaotic immune and inflammatory responses. The pattern recognition molecule, mannan-binding lectin (MBL), plays an important role in the first-line host defense against infectious agents. MBL initiates the lectin complement pathway and acts as an opsonin. Recent studies suggest that MBL also modulates inflammatory responses. We report that local responses after burn in MBL null mice differ from those f...

  17. Differential lectin binding on walls of thoraco-cervical blood vessels and lymphatics in rats.

    Science.gov (United States)

    Kagami, H; Uryu, K; Okamoto, K; Sakai, H; Kaneda, T; Sakanaka, M

    1991-08-01

    Lectin binding in the walls of large to medium-sized blood vessels and lymphatics in the rat thoraco-cervical region was examined histochemically. The tunica intima of the aorta and superficial cervical artery showed positive reactions with wheat germ agglutinin (WGA) and Concanavalin A (ConA) but not with Dolichus biflorus agglutinin (DBA). The tunica media of the aorta exhibited intense WGA binding, especially on the smooth muscle cells, but the tunica media of the superficial cervical artery did not react with the lectin. Neither ConA nor DBA bound to the tunica media of the aorta and superficial cervical artery. The tunica adventitia of both arteries contained sites binding the three lectins, although DBA reactivity declined as the vascular diameter decreased. The tunica intima of the superior vena cava and azygos vein exhibited positive WGA and ConA binding, whereas DBA binding was noted on only part of the tunica intima of the superior vena cava and not on that of the azygos vein. The tunica media and tunica adventitia were reactive for all three lectins. The WGA and ConA binding sites in the tunica adventitia showed loose networks, suggesting lectin binding on connective tissue elements interlacing among smooth muscle bundles. Lectin binding sites in the walls of lymphatics exhibited an arrangement similar to those in the walls of the veins. Moreover valves protruding into the lumen showed intense WGA and ConA binding and scattered DBA binding. Three other lectins (Ulex europaeus agglutinin, peanut agglutinin, Maclura pomifera) were examined, but they showed no reactions with the vessels. Thus, the differential binding of lectins on the walls of blood vessels and lymphatics of various sizes suggests the functional complexity of monosaccharide residues in the vascular walls. PMID:1758681

  18. Mannose-binding lectin: The Dr. Jekyll and Mr. Hyde of the innate immune system

    OpenAIRE

    Bouwman, Lee Hans

    2006-01-01

    The scope of the current thesis is to obtain insight in immunological aspects of transplantation and diabetes. This thesis underscores the current concept of collaboration between the innate and adaptive immune system by showing close interactions between both immune systems. Mannose binding lectin as a major recognition molecule of the lectin pathway and as a key protein of the immune system was studied in relation to its functional characteristics. Appreciating the Jekyll-and-Hyde character...

  19. Insecticidal Activity of Plant Lectins and Potential Application in Crop Protection

    OpenAIRE

    Maria Lígia R. Macedo; Caio F. R. Oliveira; Carolina T. Oliveira

    2015-01-01

    Lectins constitute a complex group of proteins found in different organisms. These proteins constitute an important field for research, as their structural diversity and affinity for several carbohydrates makes them suitable for numerous biological applications. This review addresses the classification and insecticidal activities of plant lectins, providing an overview of the applicability of these proteins in crop protection. The likely target sites in insect tissues, the mode of action of t...

  20. Development of recombinant Aleuria aurantia lectins with altered binding specificities to fucosylated glycans

    OpenAIRE

    Romano, Patrick R.; Mackay, Andrew; Vong, Minh; deSa, Johann; Lamontagne, Anne; Comunale, Mary Ann; Hafner, Julie; Block, Timothy; Lec, Ryszard; Mehta, Anand

    2011-01-01

    Changes in glycosylation have long been associated with disease. While there are many methods to detect changes in glycosylation, plant derived lectins are often used to determine changes on specific proteins or molecules of interest. One change in glycosylation that has been observed by us and by others is a disease or antigen associated increase in fucosylation on N-linked glycans. To measure this change, the fucose binding Aleuria aurantia lectin (AAL) is often utilized in plate and soluti...

  1. Type 2 fimbrial lectin-mediated phagocytosis of oral Actinomyces spp. by polymorphonuclear leukocytes.

    OpenAIRE

    Sandberg, A L; Mudrick, L L; Cisar, J O; Brennan, M J; Mergenhagen, S. E.; Vatter, A. E.

    1986-01-01

    Phagocytosis of Actinomyces viscosus T14V and A. naeslundii WVU45 by human polymorphonuclear leukocytes in the absence of antibody or complement was mediated by the lectin associated with the type 2 fimbriae of these bacteria. This effect was markedly enhanced by exogenous sialidase, an enzyme also secreted by these actinomyces. Since sialidase treatment of the bacteria did not result in increased phagocytosis, this enzyme presumably acts by unmasking receptors for the fimbrial lectin on phag...

  2. Recognition of immunoglobulin A1 by oral actinomyces and streptococcal lectins.

    OpenAIRE

    Ruhl, S; Sandberg, A L; Cole, M. F.; Cisar, J O

    1996-01-01

    Actinomyces naeslundii and Streptococcus gordonii, oral bacteria that possess Gal/GalNAc- and sialic acid-reactive lectins, respectively, were adherent to immobilized secretory immunoglobulin A (IgA) and two IgA1 myeloma proteins but not to two IgA2 myeloma proteins. Apparently, O-linked oligosaccharides at the hinge region of the IgA1 heavy chain are receptors for lectin-mediated adhesion of these bacteria.

  3. Mannan-binding lectin serum levels in 593 healthy Iranian children and adults.

    OpenAIRE

    2014-01-01

    Mannan-binding lectin (MBL) is a vital protein of innate immune system and has two critical functions: complement activation through the lectin pathway and opsonization. MBL deficiency has been classified as the most common inherited immunodeficiency known in humans (about 30% of the population), and is associated with predisposition to infections and high risk of some autoimmune diseases. The purpose of this study was to determine the profile of MBL serum level in Iranian healthy population ...

  4. Crystal structure of Pterocarpus angolensis lectin in complex with glucose, sucrose, and turanose.

    Science.gov (United States)

    Loris, Remy; Imberty, Anne; Beeckmans, Sonia; Van Driessche, Edilbert; Read, John S; Bouckaert, Julie; De Greve, Henri; Buts, Lieven; Wyns, Lode

    2003-05-01

    The crystal structure of the Man/Glc-specific seed lectin from Pterocarpus angolensis was determined in complex with methyl-alpha-d-glucose, sucrose, and turanose. The carbohydrate binding site contains a classic Man/Glc type specificity loop. Its metal binding loop on the other hand is of the long type, different from what is observed in other Man/Glc-specific legume lectins. Glucose binding in the primary binding site is reminiscent of the glucose complexes of concanavalin A and lentil lectin. Sucrose is found to be bound in a conformation similar as seen in the binding site of lentil lectin. A direct hydrogen bond between Ser-137(OG) to Fru(O2) in Pterocarpus angolensis lectin replaces a water-mediated interaction in the equivalent complex of lentil lectin. In the turanose complex, the binding site of the first molecule in the asymmetric unit contains the alphaGlc1-3betaFruf form of furanose while the second molecule contains the alphaGlc1-3betaFrup form in its binding site. PMID:12595543

  5. Purification of melibiose-binding lectins from two cultivars of Chinese black soybeans

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A dimeric 50 kDa melibiose-binding leetin was isolated from the seeds of the cultivar of soybean (Glycine max), called the small glossy black soybean. The isolation procedure comprised ion exchange chromatography on Q Sepharose, SP Sepharose and Mono Q followed by gel filtration on Superdex 75. The lectin was adsorbed on all three ion exchangers, and it exhibited an N-terminal sequence identical to that of soybean iectin. Of all the sugars tested, melibiose most potently inhibited the hemagglutinating activity of the lectin, which was stable between pH 3-12 and 0-70℃. The lectin evoked maximal mitogenic response at about the same molar concentration as Con A. However, the response was much weaker. The soybean iectin inhibited the activity of HIV-1 reverse transcriptase as well as the proliferation of breast cancer MCF7 cells and hepatoma HepG2 cells with an IC of 2.82 μM, 2.6 μM and 4.1 μM, respectively. There was no antifungal activity. Another lectin was isolated from a different cnltivar of soybean called little black soybean. The lectin was essentially similar to small glossy black soy-bean iectin except for a larger subunit molecular mass (31 kDa), a more potent mitogenic activity and lower thermostability. The results indicate that different cultivars of soybean produce lectins that are not identical in every aspect.

  6. Density variant glycan microarray for evaluating cross-linking of mucin-like glycoconjugates by lectins.

    Science.gov (United States)

    Godula, Kamil; Bertozzi, Carolyn R

    2012-09-26

    Interactions of mucin glycoproteins with cognate receptors are dictated by the structures and spatial organization of glycans that decorate the mucin polypeptide backbone. The glycan-binding proteins, or lectins, that interact with mucins are often oligomeric receptors with multiple ligand binding domains. In this work, we employed a microarray platform comprising synthetic glycopolymers that emulate natural mucins arrayed at different surface densities to evaluate how glycan valency and spatial separation affect the preferential binding mode of a particular lectin. We evaluated a panel of four lectins (Soybean agglutinin (SBA), Wisteria floribunda lectin (WFL), Vicia villosa-B-4 agglutinin (VVA), and Helix pomatia agglutin (HPA)) with specificity for α-N-acetylgalactosamine (α-GalNAc), an epitope displayed on mucins overexpressed in many adenocarcinomas. While these lectins possess the ability to agglutinate A(1)-blood cells carrying the α-GalNAc epitope and cross-link low valency glycoconjugates, only SBA showed a tendency to form intermolecular cross-links among the arrayed polyvalent mucin mimetics. These results suggest that glycopolymer microarrays can reveal discrete higher-order binding preferences beyond the recognition of individual glycan epitopes. Our findings indicate that glycan valency can set thresholds for cross-linking by lectins. More broadly, well-defined synthetic glycopolymers enable the integration of glycoconjugate structural and spatial diversity in a single microarray screening platform. PMID:22967056

  7. A novel antiproliferative and antifungal lectin from Amaranthus viridis Linn seeds.

    Science.gov (United States)

    Kaur, Navjot; Dhuna, Vikram; Kamboj, Sukhdev Singh; Agrewala, Javed N; Singh, Jatinder

    2006-01-01

    A lectin from the seeds of Amaranthus viridis Linn has been purified by affinity chromatography on asialofetuin-linked amino activated silica. Amaranthus viridis lectin (AVL) has a native molecular mass of 67 kDa. It is a homodimer composed of two 36.6 kDa subunits. The lectin gave a single band in non-denaturing PAGE at pH 4.5 and pH 8.3 and a single peak on HPLC size exclusion and cation exchange columns. The purified lectin was specific for both T-antigen and N-acetyl-D-lactosamine, markers for various carcinomas, in addition to N-acetyl-D-galactosamine, asialofetuin and fetuin. This lectin reacted strongly with red blood cells (RBCs) from human ABO blood groups and rat. It also reacted with rabbit, sheep, goat and guinea pig RBCs. The lectin is a glycoprotein having no metal ion requirement for its activity. Denaturing agents such as urea, thiourea and guanidine-HCl had no effect on its activity when treated for 15 minutes. AVL showed significant antiproliferative activity towards HB98 and P388D1 murine cancer cell lines. It also exerted antifungal activity against phytopathogenic fungi Botrytis cincerea and Fusarium oxysporum but not against Rhizoctonia solani, Trichoderma reesei, Alternaria solani and Fusarium graminearum. PMID:17100645

  8. Molecular Characterization of a New Lectin from the Marine Alga Ulva pertusa

    Institute of Scientific and Technical Information of China (English)

    Sheng WANG; Fu-Di ZHONG; Yong-Jiang ZHANG; Zu-Jian WU; Qi-Ying LIN; Lian-Hui XIE

    2004-01-01

    A new lectin, named UPL1, was purified from a green alga Ulvapertusa by an affinitychromatography on the bovine-thyroglobulin-Sepharose 4B column. The molecular mass of the algal lectinwas about 23 kD by SDS-PAGE, and it specifically agglutinated rabbit erythrocytes. The hemagglutinatingactivity for rabbit erythrocytes could be inhibited by bovine thyroglobulin and N-acetyl-D-glucosamine. Thelectin UPL1 required divalent cations for maintenance of its biological activity, and was heat-stable, and hadhigher activity within pH 6-8. The N-terminal amino acid sequence of the purified lectin was determined(P83209) and a set of degenerate primers were designed. The full-length cDNA of the lectin was cloned byrapid amplification of cDNA ends (RACE) method (AY433960). Sequence analysis of upl1 indicated it was1084 bp long, and encoded a premature protein of 203 amino acids. The N-terminal sequence of the matureUPL1 polypeptide started at amino acid 54 of the deduced sequence from the cDNA, indicating 53 aminoacids lost due to posttranslational modification. The primary structure of the Ulvapertusa lectin did not showamino acid sequence similarity with known plant and animal lectins. Hence, this protein may be the paradigmof a novel lectin family.

  9. Molecular Characterization of a New Lectin from the Marine Alga Ulva pertusa

    Institute of Scientific and Technical Information of China (English)

    ShengWANG; Fu-DiZHONG; Yong-JiangZHANG; Zu-JianWU; Qi-YingLIN; Lian-HuiXIE

    2004-01-01

    A new lectin, named UPL1, was purified from a green alga Ulvapertusa by an affinitychromatography on the bovine-thyroglobulin-Sepharose 4B column. The molecular mass of the algal lectinwas about 23 kD by SDS-PAGE, and it specifically agglutinated rabbit erythrocytes. The hemagglutinatingactivity for rabbit erythrocytes could be inhibited by bovine thyroglobulin and N-acetyl-D-glucosamine. Thelectin UPL1 required divalent cations for maintenance of its biological activity, and was heat-stable, and hadhigher activity within pH 6-8. The N-terminal amino acid sequence of the purified lectin was determined(P83209) and a set of degenerate primers were designed. The full-length cDNA of the lectin was cloned byrapid amplification ofcDNA ends (RACE) method (AY433960). Sequence analysis of upll indicated it was! 084 bp long, and encoded a premature protein of 203 amino acids. The N-terminal sequence of the matureUPL1 polypeptide started at amino acid 54 of the deduced sequence from the cDNA, indicating 53 aminoacids lost due to posttranslational modification. The primary structure of the Ulva pertusa lectin did not showamino acid sequence similarity with known plant and animal lectins. Hence, this protein may be the paradigmof a novel lectin family.

  10. Isolation and Characterization of Midgut Lectin From Aedes aegypti (L. (Diptera: Culicidae

    Directory of Open Access Journals (Sweden)

    Tahany Hassan Ayaad

    2015-12-01

    Full Text Available ABSTRACT The present investigation deals with the isolation and characterization of a lectin from Aedes aegypti (Ae aegypti female mid gut extract that agglutinates various mammalian red blood cells (RBCs such as human three groups A, B, and O (RH+, mouse, rat, guinea-pig, sheep and goat erythrocytes. The highest activity of both crude and isolated mid gut lectins were detected against sheep RBCs. Using (NH42 SO4 fractionation, ion-exchange and mannose-CNBr-Sepharose 6B affinity chromatography techniques, Ae. aegypti midgut lectin (Aelec was purified to homogeneity.Isoelectric focusing (IEF and reducing SDS/PAGE revealed that the isolated mid gut lectin had isoelectric point (PI of 5.90, and subunits approximate molecular weights of 35.50 and 27.35 KDa. The hemagglutination (HA of lectins were Ca2+ - independent and heat-resistant. The sugar specificity of the purified Aelec was strongly inhibited by D (+-mannose and raffinose, followed by D (+ glucose. N-acetyl-D-manosamine and N-acetyl-D-glucosamine were moderate inhibitors. None of the lectins were inhibited by the disaccharides such as galactose, lactose, trehalose (IC50 up to 200 mM or fetuin up to 1% but the glycosubstances mucin and laminarin were strong inhibitors up to very low concentrations (0.030 - 0.003%.

  11. Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling.

    Science.gov (United States)

    Shang, Yuqin; Zeng, Yun; Zeng, Yong

    2016-01-01

    Protein glycosylation is one of the key processes that play essential roles in biological functions and dysfunctions. However, progress in glycomics has considerably lagged behind genomics and proteomics, due in part to the enormous challenges in analysis of glycans. Here we present a new integrated and automated microfluidic lectin barcode platform to substantially improve the performance of lectin array for focused glycomic profiling. The chip design and flow control were optimized to promote the lectin-glycan binding kinetics and speed of lectin microarray. Moreover, we established an on-chip lectin assay which employs a very simple blocking method to effectively suppress the undesired background due to lectin binding of antibodies. Using this technology, we demonstrated focused differential profiling of tissue-specific glycosylation changes of a biomarker, CA125 protein purified from ovarian cancer cell line and different tissues from ovarian cancer patients in a fast, reproducible, and high-throughput fashion. Highly sensitive CA125 detection was also demonstrated with a detection limit much lower than the clinical cutoff value for cancer diagnosis. This microfluidic platform holds the potential to integrate with sample preparation functions to construct a fully integrated "sample-to-answer" microsystem for focused differential glycomic analysis. Thus, our technology should present a powerful tool in support of rapid advance in glycobiology and glyco-biomarker development. PMID:26831207

  12. Lectin-based glycomics: how and when was the technology born?

    Science.gov (United States)

    Hirabayashi, Jun

    2014-01-01

    Lectin-based glycomics is an emerging, comprehensive technology in the post-genome sciences. The technique utilizes a panel of lectins, which is a group of biomolecules capable of deciphering "glycocodes," with a novel platform represented by a lectin microarray. The method enables multiple glycan-lectin interaction analyses to be made so that differential glycan profiling can be performed in a rapid and sensitive manner. This approach is in clear contrast to another advanced technology, mass spectrometry, which requires prior glycan liberation. Although the lectin microarray cannot provide definitive structures of carbohydrates and their attachment sites, it gives useful clues concerning the characteristic features of glycoconjugates. These include differences not only in terminal modifications (e.g., sialic acid (Sia) linkage, types of fucosylation) but also in higher ordered structures in terms of glycan density, depth, and direction composed for both N- and O-glycans. However, before this technique began to be implemented in earnest, many other low-throughput methods were utilized in the late twentieth century. In this chapter, the author describes how the current lectin microarray technique has developed based on his personal experience. PMID:25117239

  13. The lectin pathway of complement activation is a critical component of the innate immune response to pneumococcal infection

    DEFF Research Database (Denmark)

    Ali, Youssif M; Lynch, Nicholas J; Haleem, Kashif S;

    2012-01-01

    pathways of complement in fighting streptococcal infection, little is known about the role of the lectin pathway, mainly due to the lack of appropriate experimental models of lectin pathway deficiency. We have recently established a mouse strain deficient of the lectin pathway effector enzyme mannan......The complement system plays a key role in host defense against pneumococcal infection. Three different pathways, the classical, alternative and lectin pathways, mediate complement activation. While there is limited information available on the roles of the classical and the alternative activation......-binding lectin associated serine protease-2 (MASP-2) and shown that this mouse strain is unable to form the lectin pathway specific C3 and C5 convertases. Here we report that MASP-2 deficient mice (which can still activate complement via the classical pathway and the alternative pathway) are highly susceptible...

  14. Detection of Cytotoxic Activity of Lectin on Human Colon Adenocarcinoma (Sw480 and Epithelial Cervical Carcinoma (C33-A

    Directory of Open Access Journals (Sweden)

    Mirandeli Bautista

    2011-03-01

    Full Text Available Lectins comprise a heterogeneous class of proteins that recognize the carbohydrate moieties of glycoconjugates with high specificity. Numerous studies have shown that lectins are capable of recognizing specific carbohydrate moieties displayed by malignant cells or tissues. The present work was performed to investigate the effects of tepary bean (Phaseolus acutifolius lectins on proliferation, colony formation, and alteration of DNA synthesis of human malignant cells. Tepary bean lectin showed dose dependent  effects on the inhibition of viability as well as on colony formation in two human malignant cells lines (C33-A, Sw480; By contrast, tepary bean lectin only showed significant effects on DNA synthesis on Sw480 cells. Our results provide evidence of the anti- proliferative and cytotoxic effects of the tepary bean lectins on C33-A and Sw480 cells lines.

  15. [Effect of Azospirillum lectins on the Activity of Proteolytic Enzymes and Their Inhibitors in Wheat Seedling Roots].

    Science.gov (United States)

    Alen'kina, S A; Nikitina, V E

    2015-01-01

    The lectins of associative nitrogen-fixing strains Azospirillum brasilense Sp7 and Sp245 were shown to exerte a multidirectional effect on the activity of acidic (pH 3.5), neutral (6.8), and alkaline (pH 7.8) proteinases. The lectin of the epiphytic A. brasilense Sp7 decreased proteolytic activity at all pH values, whereas the lectin of the endophytic A. brasilense Sp245 activated neutral and alkaline proteinases, while not affecting the alkaline ones. Experiments with protease inhibitors made it possible to conclude that the lectins of the studied A. brasilense strains alter the ratio between the activities of different protease types in germinating seeds. The activity of trypsin inhibitors in wheat seedling roots was found to increase in the presence of the lectins. Our results indicate a broader spectrum of effects of azospirilla lectins on the host plant organism. PMID:27169244

  16. B and C types natriuretic peptides modulate norepinephrine uptake and release in the rat hypothalamus.

    Science.gov (United States)

    Vatta, M S; Presas, M; Bianciotti, L G; Zarrabeitia, V; Fernández, B E

    1996-09-16

    We previously reported that atrial natriuretic factor (ANF) regulates catecholamine metabolism in the central nervous system. ANF, B and C types natriuretic peptides (BNP and CNP) also play a regulatory role in body fluid homeostasis, cardiovascular activity and hormonal and neuro-hormonal secretions. The aim of the present work was to investigate BNP and CNP effects on the uptake and release of norepinephrine (NE) in rat hypothalamic slices incubated in vitro. Results showed that BNP (100 nM) and CNP (1, 10 and 100 nM) enhanced total and neuronal [3H]NE uptake but did not modify non-neuronal uptake. BNP (100 nM) and CNP (1 nM) caused a rapid increase in NE uptake (1 min), which was sustained for 60 min. BNP (100 nM) did not modify the intracellular distribution of NE; however, 1 nM CNP increased the granular store and decreased the cytosolic pool of NE. BNP (100 nM) and CNP (1, 10 and 100 nM), diminished spontaneous NE release. In addition, BNP (1, 10, 100 nM) and CNP (1, 10 and 100 pM, as well as 1, 10 and 100 nM) reduced NE output induced by 25 mM KCl. These results suggest that BNP and CNP may be involved in the regulation of several central as well as peripheral physiological functions through the modulation of noradrenergic neurotransmission at the presynaptic neuronal level. Present results provide evidence to consider CNP as the brain natriuretic peptide since physiological concentrations of this peptide (pM) diminished NE evoked release. PMID:8897640

  17. Dexamethasone stimulates expression of C-type Natriuretic Peptide in chondrocytes

    Directory of Open Access Journals (Sweden)

    Beier Frank

    2006-11-01

    Full Text Available Abstract Background Growth of endochondral bones is regulated through the activity of cartilaginous growth plates. Disruption of the physiological patterns of chondrocyte proliferation and differentiation – such as in endocrine disorders or in many different genetic diseases (e.g. chondrodysplasias – generally results in dwarfism and skeletal defects. For example, glucocorticoid administration in children inhibits endochondral bone growth, but the molecular targets of these hormones in chondrocytes remain largely unknown. In contrast, recent studies have shown that C-type Natriuretic Peptide (CNP is an important anabolic regulator of cartilage growth, and loss-of-function mutations in the human CNP receptor gene cause dwarfism. We asked whether glucocorticoids could exert their activities by interfering with the expression of CNP or its downstream signaling components. Methods Primary mouse chondrocytes in monolayer where incubated with the synthetic glucocorticoid Dexamethasone (DEX for 12 to 72 hours. Cell numbers were determined by counting, and real-time PCR was performed to examine regulation of genes in the CNP signaling pathway by DEX. Results We show that DEX does influence expression of key genes in the CNP pathway. Most importantly, DEX significantly increases RNA expression of the gene encoding CNP itself (Nppc. In addition, DEX stimulates expression of Prkg2 (encoding cGMP-dependent protein kinase II and Npr3 (natriuretic peptide decoy receptor genes. Conversely, DEX was found to down-regulate the expression of the gene encoding its receptor, Nr3c1 (glucocorticoid receptor, as well as the Npr2 gene (encoding the CNP receptor. Conclusion Our data suggest that the growth-suppressive activities of DEX are not due to blockade of CNP signaling. This study reveals a novel, unanticipated relationship between glucocorticoid and CNP signaling and provides the first evidence that CNP expression in chondrocytes is regulated by endocrine

  18. Absence of C-type virus production in human leukemic B cell, T cell and null cell lines.

    Directory of Open Access Journals (Sweden)

    Ogura,Hajime

    1978-06-01

    Full Text Available Electron microscope observation of cultured human leukemic B cell, T cell and null cell lines and reverse transcriptase assay of the culture supernatants were all negative for the presence of C-type virus. Bat cell line, which propagates primate C-type viruses well, was cocultivated with the human leukemic cell lines, in the hope of amplification of virus if present. Three weeks after mixed culture, the culture supernatants were again examined for reverse transcriptase activity and the cells were tested for syncytia formation by cocultivation with rat XC, human KC and RSb cell lines. All these tests, except for the positive control using a simian sarcoma virus, were negative, suggesting that no C-type was produced from these human leukemic cell lines.

  19. The Breast Cancer-Associated Glycoforms of MUC1, MUC1-Tn and sialyl-Tn, Are Expressed in COSMC Wild-Type Cells and Bind the C-Type Lectin MGL

    DEFF Research Database (Denmark)

    Beatson, Richard; Maurstad, Gjertrud; Picco, Gianfranco; Arulappu, Appitha; Coleman, Julia; Wandell, Hans H; Clausen, Henrik; Mandel, Ulla; Taylor-Papadimitriou, Joyce; Sletmoen, Marit; Burchell, Joy M

    2015-01-01

    close proximity to extended glycan T (Galβ1,3GalNAc) the addition of Gal to the GalNAc being catalysed by the T synthase. The presence of active T synthase suggests that loss of the private chaperone for T synthase, COSMC, does not explain the expression of Tn and STn in breast cancer cells. We show......, DC apoptosis and inhibition of NK activity. As engagement of MGL in the absence of TLR triggering may lead to anergy, the binding of MUC1-STn to MGL may be in part responsible for some of the characteristics of STn expressing tumours....

  20. 犬钩虫C型凝集素AcaCTL-1基因的克隆及表达%Cloning and expression of a C-type lectin(AcaCTL-1)gene from Ancylostoma caninum

    Institute of Scientific and Technical Information of China (English)

    邓莉; 金娴; 何庆丰; 许琴英; 吴亚敏; 彭礼飞

    2010-01-01

    目的 分离鉴定犬钩虫C型凝集素AcaCTL-1基因,并在大肠埃希菌中表达.方法 根据EST序列设计引物,用RT-PCR及3,(RACE技术扩增获得AcaCTL-1伞长cDNA序列,并对其进行初步生物信息学分析;将AcaCTL-l成熟肽编码序列克隆、连接到表达载体pET32a.构建重组质粒pET32a/AcaCTL-1;在大肠埃希菌BL21(DE3)中诱导表达重组蛋白,SDS-PAGE分析表达情况,Ni亲和层析纯化可溶性蛋白.结果 成功克隆了AcaCTL-1全长cDNA序列,其最大开放阅读框由525 bp组成,预测编码174个氨基酸残基组成的蛋白,含17个氨基酸组成的信号肽,为C型凝集索家族蛋白成员;构建pET32a/AcaCTL-1莺组质粒,并在大肠埃希菌中高效表达.表达产物多以包涵体形式存在,小部分为可溶性蛋白.结论 成功克隆并表达了犬钩虫C型凝集素AcaCTL-基因,为进一步了解AcaCTL-1的功能奠定了基础.

  1. Re-evaluation of the involvement of NK cells and C-type lectin-like NK receptors in modulation of immune responses by multivalent GlcNAc-terminated oligosaccharides

    Czech Academy of Sciences Publication Activity Database

    Grobárová, Valeria; Benson, Veronika; Rozbeský, Daniel; Novák, Petr; Černý, O.

    2013-01-01

    Roč. 156, 1-2 (2013), s. 110-117. ISSN 0165-2478 R&D Projects: GA MŠk ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : Anti-tumor immunity * Carbohydrate dendrimer * NK cells Subject RIV: EC - Immunology Impact factor: 2.367, year: 2013

  2. A Comparative Study of Lectin Affinity Based Plant N-Glycoproteome Profiling Using Tomato Fruit as a Model*

    OpenAIRE

    Ruiz-May, Eliel; Hucko, Simon; Kevin J. Howe; Zhang, Sheng; Sherwood, Robert W.; Thannhauser, Theodore W; Rose, Jocelyn K. C.

    2013-01-01

    Lectin affinity chromatography (LAC) can provide a valuable front-end enrichment strategy for the study of N-glycoproteins and has been used to characterize a broad range eukaryotic N-glycoproteomes. Moreover, studies with mammalian systems have suggested that the use of multiple lectins with different affinities can be particularly effective. A multi-lectin approach has also been reported to provide a significant benefit for the analysis of plant N-glycoproteins; however, it has yet to be de...

  3. The lectin pathway of complement activation is a critical component of the innate immune response to pneumococcal infection.

    Directory of Open Access Journals (Sweden)

    Youssif M Ali

    Full Text Available The complement system plays a key role in host defense against pneumococcal infection. Three different pathways, the classical, alternative and lectin pathways, mediate complement activation. While there is limited information available on the roles of the classical and the alternative activation pathways of complement in fighting streptococcal infection, little is known about the role of the lectin pathway, mainly due to the lack of appropriate experimental models of lectin pathway deficiency. We have recently established a mouse strain deficient of the lectin pathway effector enzyme mannan-binding lectin associated serine protease-2 (MASP-2 and shown that this mouse strain is unable to form the lectin pathway specific C3 and C5 convertases. Here we report that MASP-2 deficient mice (which can still activate complement via the classical pathway and the alternative pathway are highly susceptible to pneumococcal infection and fail to opsonize Streptococcus pneumoniae in the none-immune host. This defect in complement opsonisation severely compromises pathogen clearance in the lectin pathway deficient host. Using sera from mice and humans with defined complement deficiencies, we demonstrate that mouse ficolin A, human L-ficolin, and collectin 11 in both species, but not mannan-binding lectin (MBL, are the pattern recognition molecules that drive lectin pathway activation on the surface of S. pneumoniae. We further show that pneumococcal opsonisation via the lectin pathway can proceed in the absence of C4. This study corroborates the essential function of MASP-2 in the lectin pathway and highlights the importance of MBL-independent lectin pathway activation in the host defense against pneumococci.

  4. Crystallization and preliminary X-ray diffraction analysis of HML, a lectin from the red marine alga Hypnea musciformis

    International Nuclear Information System (INIS)

    The crystallization and preliminary X-ray diffraction analysis of a red marine alga lectin isolated from H. musciformis is reported. HML, a lectin from the red marine alga Hypnea musciformis, defines a novel lectin family. Orthorhombic crystals of HML belonging to space group P212121 grew within three weeks at 293 K using the hanging-drop vapour-diffusion method. A complete data set was collected at 2.4 Å resolution. HML is the first marine alga lectin to be crystallized

  5. A single-molecule force spectroscopy study of the interactions between lectins and carbohydrates on cancer and normal cells

    Science.gov (United States)

    Zhao, Weidong; Cai, Mingjun; Xu, Haijiao; Jiang, Junguang; Wang, Hongda

    2013-03-01

    The interaction forces between carbohydrates and lectins were investigated by single-molecule force spectroscopy on both cancer and normal cells. The binding kinetics was also studied, which shows that the carbohydrate-lectin complex on cancer cells is less stable than that on normal cells.The interaction forces between carbohydrates and lectins were investigated by single-molecule force spectroscopy on both cancer and normal cells. The binding kinetics was also studied, which shows that the carbohydrate-lectin complex on cancer cells is less stable than that on normal cells. Electronic supplementary information (ESI) available: Experimental details. See DOI: 10.1039/c3nr00553d

  6. Histological and lectin histochemical studies on the main and accessory olfactory bulbs in the Japanese striped snake, Elaphe quadrivirgata.

    Science.gov (United States)

    Kondoh, Daisuke; Wada, Akimi; Endo, Daisuke; Nakamuta, Nobuaki; Taniguchi, Kazuyuki

    2013-01-01

    The main and accessory olfactory bulbs were examined by histological methods and lectin histochemistry in the Japanese striped snake. As the results, the histological properties are similar between the main olfactory bulb and the accessory olfactory bulb. In lectin histochemistry, 21 lectins used in this study showed similar binding patterns in the main olfactory bulb and the accessory olfactory bulb. In detail, 15 lectins stained these olfactory bulbs with similar manner, and 6 lectins did not stain them at all. Two lectins, Lycopersicon esculentum lectin (LEL) and Solanum tuberosum lectin (STL), stained the nerve and glomerular layers and did not stain any other layers in both olfactory bulbs. Four lectins, Soybean agglutinin (SBA), Vicia villosa agglutinin (VVA), Peanut agglutinin (PNA) and Phaseolus vulgaris agglutinin-L (PHA-L) stained the nerve and glomerular layers more intensely than other layers in both olfactory bulbs. In addition, VVA showed the dot-like stainings in the glomeruli of both olfactory bulbs. These findings suggest that the degree of development and the properties of glycoconjugates are similar between the main olfactory bulb and the accessory olfactory bulb in the Japanese striped snake. PMID:23257605

  7. PURIFICATION AND CHARACTERIZATION OF A NEW MANNOSE-SPECIFIC LECTIN FROM Hyacinthella аcutiloba K. Perss.

    Directory of Open Access Journals (Sweden)

    V. O. Antonyuk

    2013-06-01

    Full Text Available A new lectin with 6.5 mg/kg yield was purified from fresh bulb Hyacinthella аcutiloba K. Perss. by combination of affinity chromathography on yeast mannan and by ion exchange chromatography on DEAE-toyopearl. The best lectin inhibitor among tested mono- and disaccharides was D-turanose (Glcp ?1-3 Fruf. Very powerful inhibitor of lectin activity was yeast mannan. The lectin revealed weak affinity to ?-methyl-Dmannoside, D-fructose and 2-acetamido-D-galactopyranoside. The lectin interacted also with pig liver glycogen, starch and mannose-containing glycoproteins (ovoalbumin, ovomucoid and calf thyroglobulin, but don’t interacted with horsera dish peroxidase and calf intestine alkaline phosphatase. According to electrophoresis, in 20% SDS-PAAG containing SDS-Na the lectin consists with subunits of molecular weight 12 kDa. Molecular weight of the lectin is 48 kDa according to gelchromatography on Toyopearl HW-55. The lectin agglutinated best of all rabbit erythrocytes and worse agglutinated guinea-pig, very weak — rat erythrocytes and did not agglutinate human and goat erythrocytes. After dialysis against 1% EDTA sodium salt solution the lectin did not lose hemaglutinating activity and endured heating to +65 °C during one hour.

  8. Crystallization and preliminary X-ray diffraction analysis of HML, a lectin from the red marine alga Hypnea musciformis

    Energy Technology Data Exchange (ETDEWEB)

    Nagano, Celso S.; Gallego del Sol, Francisca [Instituto de Biomedicina de Valencia, CSIC, Valencia (Spain); Cavada, Benildo S.; Nascimento, Kyria Santiago Do [Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Ceará, Fortaleza, CE 60451-970 (Brazil); Nunes, Eudismar Vale; Sampaio, Alexandre H. [Laboratorio de Bioquímica Marinha, Departamento de Engenharia de Pesca, Universidade Federal do Ceará, Fortaleza, CE 60451-970 (Brazil); Calvete, Juan J., E-mail: jcalvete@ibv.csic.es [Instituto de Biomedicina de Valencia, CSIC, Valencia (Spain)

    2005-11-01

    The crystallization and preliminary X-ray diffraction analysis of a red marine alga lectin isolated from H. musciformis is reported. HML, a lectin from the red marine alga Hypnea musciformis, defines a novel lectin family. Orthorhombic crystals of HML belonging to space group P2{sub 1}2{sub 1}2{sub 1} grew within three weeks at 293 K using the hanging-drop vapour-diffusion method. A complete data set was collected at 2.4 Å resolution. HML is the first marine alga lectin to be crystallized.

  9. A lectin from Platypodium elegans with unusual specificity and affinity for asymmetric complex N-glycans.

    Science.gov (United States)

    Benevides, Raquel Guimarães; Ganne, Géraldine; Simões, Rafael da Conceição; Schubert, Volker; Niemietz, Mathäus; Unverzagt, Carlo; Chazalet, Valérie; Breton, Christelle; Varrot, Annabelle; Cavada, Benildo Sousa; Imberty, Anne

    2012-07-27

    Lectin activity with specificity for mannose and glucose has been detected in the seed of Platypodium elegans, a legume plant from the Dalbergieae tribe. The gene of Platypodium elegans lectin A has been cloned, and the resulting 261-amino acid protein belongs to the legume lectin family with similarity with Pterocarpus angolensis agglutinin from the same tribe. The recombinant lectin has been expressed in Escherichia coli and refolded from inclusion bodies. Analysis of specificity by glycan array evidenced a very unusual preference for complex type N-glycans with asymmetrical branches. A short branch consisting of one mannose residue is preferred on the 6-arm of the N-glycan, whereas extensions by GlcNAc, Gal, and NeuAc are favorable on the 3-arm. Affinities have been obtained by microcalorimetry using symmetrical and asymmetrical Asn-linked heptasaccharides prepared by the semi-synthetic method. Strong affinity with K(d) of 4.5 μm was obtained for both ligands. Crystal structures of Platypodium elegans lectin A complexed with branched trimannose and symmetrical complex-type Asn-linked heptasaccharide have been solved at 2.1 and 1.65 Å resolution, respectively. The lectin adopts the canonical dimeric organization of legume lectins. The trimannose bridges the binding sites of two neighboring dimers, resulting in the formation of infinite chains in the crystal. The Asn-linked heptasaccharide binds with the 6-arm in the primary binding site with extensive additional contacts on both arms. The GlcNAc on the 6-arm is bound in a constrained conformation that may rationalize the higher affinity observed on the glycan array for N-glycans with only a mannose on the 6-arm. PMID:22692206

  10. A Lectin from Platypodium elegans with Unusual Specificity and Affinity for Asymmetric Complex N-Glycans*

    Science.gov (United States)

    Benevides, Raquel Guimarães; Ganne, Géraldine; Simões, Rafael da Conceição; Schubert, Volker; Niemietz, Mathäus; Unverzagt, Carlo; Chazalet, Valérie; Breton, Christelle; Varrot, Annabelle; Cavada, Benildo Sousa; Imberty, Anne

    2012-01-01

    Lectin activity with specificity for mannose and glucose has been detected in the seed of Platypodium elegans, a legume plant from the Dalbergieae tribe. The gene of Platypodium elegans lectin A has been cloned, and the resulting 261-amino acid protein belongs to the legume lectin family with similarity with Pterocarpus angolensis agglutinin from the same tribe. The recombinant lectin has been expressed in Escherichia coli and refolded from inclusion bodies. Analysis of specificity by glycan array evidenced a very unusual preference for complex type N-glycans with asymmetrical branches. A short branch consisting of one mannose residue is preferred on the 6-arm of the N-glycan, whereas extensions by GlcNAc, Gal, and NeuAc are favorable on the 3-arm. Affinities have been obtained by microcalorimetry using symmetrical and asymmetrical Asn-linked heptasaccharides prepared by the semi-synthetic method. Strong affinity with Kd of 4.5 μm was obtained for both ligands. Crystal structures of Platypodium elegans lectin A complexed with branched trimannose and symmetrical complex-type Asn-linked heptasaccharide have been solved at 2.1 and 1.65 Å resolution, respectively. The lectin adopts the canonical dimeric organization of legume lectins. The trimannose bridges the binding sites of two neighboring dimers, resulting in the formation of infinite chains in the crystal. The Asn-linked heptasaccharide binds with the 6-arm in the primary binding site with extensive additional contacts on both arms. The GlcNAc on the 6-arm is bound in a constrained conformation that may rationalize the higher affinity observed on the glycan array for N-glycans with only a mannose on the 6-arm. PMID:22692206

  11. Diagnosis of myocardial infarction based on lectin-induced erythrocyte agglutination: a feasibility study

    Science.gov (United States)

    Bocsi, József; Nieschke, Kathleen; Mittag, Anja; Reichert, Thomas; Laffers, Wiebke; Marecka, Monika; Pierzchalski, Arkadiusz; Piltz, Joachim; Esche, Hans-Jürgen; Wolf, Günther; Dähnert, Ingo; Baumgartner, Adolf; Tarnok, Attila

    2014-03-01

    Myocardial infarction (MI) is an acute life-threatening disease with a high incidence worldwide. Aim of this study was to test lectin-carbohydrate binding-induced red blood cell (RBC) agglutination as an innovative tool for fast, precise and cost effective diagnosis of MI. Five lectins (Ricinus communis agglutinin (RCA), Phaseolus vulgaris erythroagglutinin (PHA), Datura stramonium agglutinin (DSA), Artocarpus agglutinin (ArA), Triticum agglutinin (TA)) were tested for ability to differentiate between agglutination characteristics in patients with MI (n = 101) or angina pectoris without MI (AP) (n = 34) and healthy volunteers (HV) as control (n =68) . RBC agglutination was analyzed by light absorbance of a stirred RBC suspension in the green to red light spectrum in an agglutimeter (amtec, Leipzig, Germany) for 15 min after lectin addition. Mean cell count in aggregates was estimated from light absorbance by a mathematical model. Each lectin induced RBC agglutination. RCA led to the strongest RBC agglutination (~500 RBCs/aggregate), while the others induced substantially slower agglutination and lead to smaller aggregate sizes (5-150 RBCs/aggregate). For all analyzed lectins the lectin-induced RBC agglutination of MI or AP patients was generally higher than for HV. However, only PHA induced agglutination that clearly distinguished MI from HV. Variance analysis showed that aggregate size after 15 min. agglutination induced by PHA was significantly higher in the MI group (143 RBCs/ aggregate) than in the HV (29 RBC-s/aggregate, p = 0.000). We hypothesize that pathological changes during MI induce modification of the carbohydrate composition on the RBC membrane and thus modify RBC agglutination. Occurrence of carbohydrate-lectin binding sites on RBC membranes provides evidence about MI. Due to significant difference in the rate of agglutination between MI > HV the differentiation between these groups is possible based on PHA-induced RBC-agglutination. This novel assay

  12. Mannose-Binding Lectin Serum Levels in Patients With Candiduria

    Science.gov (United States)

    Moslem, Maryam; Zarei Mahmoudabadi, Ali; Fatahinia, Mahnaz; Kheradmand, Alireza

    2015-01-01

    Background: Candida species are normal mycoflora of human body which are capable to cause urinary tract infection (UTI). Mannose-binding lectin (MBL) is a kind of innate immune system and decreasing plasma levels of MBL may disrupt the natural immune response and increase susceptibility to infections. Objectives: The aim of the present study was to assess MBL in the serum of patients with candiduria and compare them with control. Patients and Methods: The blood and urine samples were collected from 335 patients (hospitalized in Golestan hospital, Ahvaz) using standard methods and the growing colonies on CHROMagar were identified using routine diagnostic tests. MBL activity in the serum of 45 patients with candiduria and 45 controls was measured using Eastbiopharm enzyme-linked immunosorbent assay (ELISA) kit. Results: In this study, 45 (13.4 %) urine samples were positive for Candida species (17 males and 28 females). The most common isolated yeast was Candida albicans (34%), followed by C. glabrata (32.1%), C. tropicalis (9.4%), other Candida species (22.6%), and Rhodotorula species (1.9%). The mean serum levels of MBL were 0.85 ± 0.01 ng/mL and 1.02 ± 0.03 ng/mL among candiduric patients and controls, respectively, and there was no significant difference between the two groups (P = 0.6). Conclusions: Our results showed that there was no significant relationship between MBL serum levels and candiduria. PMID:26870314

  13. Thermoresponsive diblock glycopolymer by RAFT polymerization for lectin recognition.

    Science.gov (United States)

    Sun, Kan; Xu, Muru; Zhou, Kaichun; Nie, Huali; Quan, Jing; Zhu, Limin

    2016-11-01

    A thermoresponsive double-hydrophilic diblock glycopolymer, poly(diethyl- eneglycol methacrylate)-block-poly(6-O-vinyladipoyl-d-glucose) (PDEGMA-b-POVAG), was successfully prepared by a combination of enzymatic synthesis and reversible addition-fragment chain transfer (RAFT) polymerization protocols using poly(diethyl- eneglycol methacrylate) (PDEGMA) as macro-RAFT agent. The block glycopolymer was characterized by (1)H NMR and GPC. UV-vis, DLS and TEM studies revealed that the glycopolymer PDEGMA-b-POVAG was thermoresponsive with LCST at 31.0°C, and was able to self-assemble into spherical micelles of various sizes in aqueous solution. The glucose pendants in the glycopolymer could interact with the lectin Concanavalin A (Con A), the average hydrodynamic diameters of glycopolymer micelles increased to 170nm from 110nm after recognizing Con A. The diblock glycopolymer micelles have excellent biocompatibility with pig iliac endothelial cells, as measured using the MTT assay, but micelles loaded with Con A could be used to induce apoptosis in human hepatoma SMMC-7721 cells. PMID:27524009

  14. [Immunoactive action of mistletoe lectin-1 in relation to dose].

    Science.gov (United States)

    Beuth, J; Ko, H L; Tunggal, L; Buss, G; Jeljaszewicz, J; Steuer, M K; Pulverer, G

    1994-11-01

    Galactoside-specific mistletoe lectin-1 (ML-1) was isolated by affinity chromatography from proprietary mistletoe extract and checked in BALB/c-mice for its immunoactive potency. To investigate the optimal immunomodulating dosage, ML-1 (0.5, 1.0, 2.5, 5.0 ng/kg body weight, b.w.) was subcutaneously administered for three subsequent days followed by another injection 48 h later. These studies proved that injections of 1 ng ML-1/kg b.w. induced optimal immunomodulation, since thymocyte proliferation, maturation and emigration were significantly enhanced in this murine model as compared to non-treated control mice. Further on, counts of peripheral blood lymphocytes and monocytes as well as expression of relevant activation markers on these cells revealed significant increases after ML-1 (1 ng/kg b.w.) administration. However, increase of cell counts and activity of peritoneal macrophages were less pronounced but still statistically significant for this ML-1 concentration. Determination of immune responses after low dose ML-1 treatment (0.5 ng/kg b.w.) presented relevant (partly statistically significant) increases, too. However, high dose ML-1 treatment (2.5, 5.0 ng/kg b.w.) did not enhance (but suppress) relevant immune functions. For future clinical/therapeutical treatment strategies, ML-1 dosages ranging from 0.5-1.0 ng/kg b.w. may be supposed to be optimal. PMID:7848341

  15. Ultrasensitive impedimetric lectin based biosensor for glycoproteins containing sialic acid

    Science.gov (United States)

    Bertok, Tomas; Gemeiner, Pavol; Mikula, Milan; Gemeiner, Peter; Tkac, Jan

    2016-01-01

    We report on an ultrasensitive label-free lectin-based impedimetric biosensor for the determination of the sialylated glycoproteins fetuin and asialofetuin. A sialic acid binding agglutinin from Sambucus nigra I was covalently immobilised on a mixed self-assembled monolayer (SAM) consisting of 11-mercaptoundecanoic acid and 6-mercaptohexanol. Poly(vinyl alcohol) was used as a blocking agent. The sensor layer was characterised by atomic force microscopy, electrochemical impedance spectroscopy and X-ray photoelectron spectroscopy. The biosensor exhibits a linear range that spans 7 orders of magnitude for both glycoproteins, with a detection limit as low as 0.33 fM for fetuin and 0.54 fM for asialofetuin. We also show, by making control experiments with oxidised asialofetuin, that the biosensor is capable of quantitatively detecting changes in the fraction of sialic acid on glycoproteins. We conclude that this work lays a solid foundation for future applications of such a biosensor in terms of the diagnosis of diseases such as chronic inflammatory rheumatoid arthritis, genetic disorders and cancer, all of which are associated with aberrant glycosylation of protein biomarkers.

  16. Mannose-Binding Lectin Serum Levels in Patients With Candiduria

    Directory of Open Access Journals (Sweden)

    Moslem

    2015-12-01

    Full Text Available Background Candida species are normal mycoflora of human body which are capable to cause urinary tract infection (UTI. Mannose-binding lectin (MBL is a kind of innate immune system and decreasing plasma levels of MBL may disrupt the natural immune response and increase susceptibility to infections. Objectives The aim of the present study was to assess MBL in the serum of patients with candiduria and compare them with control. Patients and Methods The blood and urine samples were collected from 335 patients (hospitalized in Golestan hospital, Ahvaz using standard methods and the growing colonies on CHROMagar were identified using routine diagnostic tests. MBL activity in the serum of 45 patients with candiduria and 45 controls was measured using Eastbiopharm enzyme-linked immunosorbent assay (ELISA kit. Results In this study, 45 (13.4 % urine samples were positive for Candida species (17 males and 28 females. The most common isolated yeast was Candida albicans (34%, followed by C. glabrata (32.1%, C. tropicalis (9.4%, other Candida species (22.6%, and Rhodotorula species (1.9%. The mean serum levels of MBL were 0.85 ± 0.01 ng/mL and 1.02 ± 0.03 ng/mL among candiduric patients and controls, respectively, and there was no significant difference between the two groups (P = 0.6. Conclusions Our results showed that there was no significant relationship between MBL serum levels and candiduria.

  17. Scaffold Optimisation of Tetravalent Antagonists of the Mannose Binding Lectin.

    Science.gov (United States)

    Goti, Giulio; Palmioli, Alessandro; Stravalaci, Matteo; Sattin, Sara; De Simoni, Maria-Grazia; Gobbi, Marco; Bernardi, Anna

    2016-03-01

    Antagonists of mannose binding lectin (MBL) have shown a protective role against brain reperfusion damage after acute ischemic stroke. Here we describe the design and streamlined synthesis of glycomimetic MBL antagonists based on a new tetravalent dendron scaffold. The dendron was developed by optimisation of a known polyester structure previously demonstrated to be very efficient for ligand presentation to MBL. Replacement of a labile succinyl ester bond with a more robust amide functionality, use of a longer and more hydrophilic linker, fast modular synthesis and orthogonal functionalisation at the focal point are the main features of the new scaffold. The glycoconjugate constructs become stable to silica gel chromatography and to water solutions at physiological pH, while preserving water solubility and activity in an SPR assay against the murine MBL-C isoform. Higher-order constructs were easily assembled, as demonstrated by the synthesis of a 16-valent dendrimer, which leads to two orders of magnitude increase in activity over the tetravalent version against MBL-C. PMID:26696414

  18. Entamoeba histolytica: Adhesins and Lectins in the Trophozoite Surface

    Directory of Open Access Journals (Sweden)

    Magdalena Aguirre García

    2015-02-01

    Full Text Available Entamoeba histolytica is the causative agent of amebiasis in humans and is responsible for 100,000 deaths annually, making it the third leading cause of death due to a protozoan parasite. Pathogenesis appears to result from the potent cytotoxic activity of the parasite, which kills host cells within minutes. Although the mechanism is unknown, it is well established to be contact-dependent. The life cycle of the parasite alternates with two forms: the resistant cyst and the invasive trophozoite. The adhesive interactions between the parasite and surface glycoconjugates of host cells, as well as those lining the epithelia, are determinants for invasion of human tissues, for its cytotoxic activity, and finally for the outcome of the disease. In this review we present an overview of the information available on the amebic lectins and adhesins that are responsible of those adhesive interactions and we also refer to their effect on the host immune response. Finally, we present some concluding remarks and perspectives in the field.

  19. Destabilization of pea lectin by substitution of a single amino acid in a surface loop.

    Science.gov (United States)

    Hoedemaeker, F J; van Eijsden, R R; Díaz, C L; de Pater, B S; Kijne, J W

    1993-09-01

    Legume lectins are considered to be antinutritional factors (ANF) in the animal feeding industry. Inactivation of ANF is an important element in processing of food. In our study on the stability of Pisum sativum L. lectin (PSL), a conserved hydrophobic amino acid (Val103) in a surface loop was replaced with alanine. The mutant lectin, PSL V103A, showed a decrease in unfolding temperature (Tm) by some 10 degrees C in comparison with wild-type (wt) PSL, and the denaturation energy (delta H) is only about 55% of that of wt PSL. Replacement of an adjacent amino acid (Phe104) with alanine did not result in a significant difference in stability in comparison with wt PSL. Both mutations did not change the sugar-binding properties of the lectin, as compared with wt PSL and with PSL from pea seeds, at ambient temperatures. The double mutant, PSL V103A/F104A, was produced in Escherichia coli, but could not be isolated in an active (i.e. sugar-binding) form. Interestingly, the mutation in PSL V103A reversibly affected sugar-binding at 37 degrees C, as judged from haemagglutination assays. These results open the possibility of production of lectins that are active in planta at ambient temperatures, but are inactive and possibly non-toxic at 37 degrees C in the intestines of mammals. PMID:8400124

  20. Toward a structure-based comprehension of the lectin pathway of complement

    DEFF Research Database (Denmark)

    Kjaer, Troels R; Thiel, Steffen; Andersen, Gregers R

    2013-01-01

    To initiate the lectin pathway of complement pattern recognition molecules bind to surface-linked carbohydrates or acetyl groups on pathogens or damaged self-tissue. This leads to activation of the serine proteases MASP-1 and MASP-2 resulting in deposition of C4 on the activator and assembly of t...... concerning the lectin pathway proteins and derive overall models for their glycan bound complexes. These models are used to discuss cis- versus trans-activation of MASP proteases and the geometry of C4 deposition occurring on glycans in the lectin pathway......To initiate the lectin pathway of complement pattern recognition molecules bind to surface-linked carbohydrates or acetyl groups on pathogens or damaged self-tissue. This leads to activation of the serine proteases MASP-1 and MASP-2 resulting in deposition of C4 on the activator and assembly of the...... activation progresses from pattern recognition to convertase assembly. Furthermore, atomic structures derived by crystallography or solution scattering of most proteins acting in the lectin pathway and two key complexes have become available. Here we integrate the current functional and structural knowledge...

  1. Effect of Chum Salmon Egg Lectin on Tight Junctions in Caco-2 Cell Monolayers

    Directory of Open Access Journals (Sweden)

    Ryo Nemoto

    2015-05-01

    Full Text Available The effect of a chum salmon egg lectin (CSL3 on tight junction (TJ of Caco-2 cell monolayers was investigated. The lectin opened TJ as indicated by the decrease of the transepithelial electrical resistance (TER value and the increase of the permeation of lucifer yellow, which is transported via the TJ-mediated paracellular pathway. The effects of CSL3 were inhibited by the addition of 10 mM L-rhamnose or D-galactose which were specific sugars for CSL3. The lectin increased the intracellular Ca2+ of Caco-2 cell monolayers, that could be inhibited by the addition of L-rhamnose. The fluorescence immunostaining of β-actin in Caco-2 cell monolayers revealed that the cytoskeleton was changed by the CSL3 treatment, suggesting that CSL3 depolymerized β-actin to cause reversible TJ structural and functional disruption. Although Japanese jack bean lectin and wheat germ lectin showed similar effects in the decrease of the TER values and the increase of the intracellular Ca2+, they could not be inhibited by the same concentrations of simple sugars, such as D-glucose and N-acetyl-D-glucosamine.

  2. Effect of chum salmon egg lectin on tight junctions in Caco-2 cell monolayers.

    Science.gov (United States)

    Nemoto, Ryo; Yamamoto, Shintaro; Ogawa, Tomohisa; Naude, Ryno; Muramoto, Koji

    2015-01-01

    The effect of a chum salmon egg lectin (CSL3) on tight junction (TJ) of Caco-2 cell monolayers was investigated. The lectin opened TJ as indicated by the decrease of the transepithelial electrical resistance (TER) value and the increase of the permeation of lucifer yellow, which is transported via the TJ-mediated paracellular pathway. The effects of CSL3 were inhibited by the addition of 10 mM L-rhamnose or D-galactose which were specific sugars for CSL3. The lectin increased the intracellular Ca2+ of Caco-2 cell monolayers, that could be inhibited by the addition of L-rhamnose. The fluorescence immunostaining of β-actin in Caco-2 cell monolayers revealed that the cytoskeleton was changed by the CSL3 treatment, suggesting that CSL3 depolymerized β-actin to cause reversible TJ structural and functional disruption. Although Japanese jack bean lectin and wheat germ lectin showed similar effects in the decrease of the TER values and the increase of the intracellular Ca2+, they could not be inhibited by the same concentrations of simple sugars, such as D-glucose and N-acetyl-D-glucosamine. PMID:25951005

  3. Structural-functional insights and studies on saccharide binding of Sophora japonica seed lectin.

    Science.gov (United States)

    Yadav, Priya; Shahane, Ganesh; Ramasamy, Sureshkumar; Sengupta, Durba; Gaikwad, Sushama

    2016-10-01

    Functional and conformational transitions of the Sophora japonica seed lectin (SJL) were studied in detail using bioinformatics and biophysical tools. Homology model of the lectin displayed all the characteristics of the legume lectin monomer and the experimental observations correlated well with the structural information. In silico studies were performed by protein-ligand docking, calculating the respective binding energies and the residues involved in the interactions were derived from LigPlot(+) analysis. Fluorescence titrations showed three times higher affinity of T-antigen disaccharide than N-acetyl galactosamine (GalNAc) towards SJL indicating extended sugar binding site of the lectin. Thermodynamic parameters of T-antigen binding to SJL indicated the process to be endothermic and entropically driven while those of GalNAc showed biphasic process. SDS-PAGE showed post-translationally modified homotetrameric species of the lectin under native conditions. In presence of guanidine hydrochloride (0.5-5.0M), the tetramer first dissociated into dimers followed by unfolding of the protein as indicated by size exclusion chromatography, fluorescence and CD spectroscopy. Different structural rearrangements were observed during thermal denaturation of SJL at physiological pH 7.2, native pH 8.5 and molten globule inducing pH 1.0. Topological information revealed by solute quenching studies at respective pH indicated differential hydrophobic environment and charge density around tryptophan residues. PMID:27185070

  4. Glycan heterogeneity on gold nanoparticles increases lectin discrimination capacity in label-free multiplexed bioassays.

    Science.gov (United States)

    Otten, Lucienne; Vlachou, Denise; Richards, Sarah-Jane; Gibson, Matthew I

    2016-07-21

    The development of new analytical tools as point-of-care biosensors is crucial to combat the spread of infectious diseases, especially in the context of drug-resistant organisms, or to detect biological warfare agents. Glycan/lectin interactions drive a wide range of recognition and signal transduction processes within nature and are often the first site of adhesion/recognition during infection making them appealing targets for biosensors. Glycosylated gold nanoparticles have been developed that change colour from red to blue upon interaction with carbohydrate-binding proteins and may find use as biosensors, but are limited by the inherent promiscuity of some of these interactions. Here we mimic the natural heterogeneity of cell-surface glycans by displaying mixed monolayers of glycans on the surface of gold nanoparticles. These are then used in a multiplexed, label-free bioassay to create 'barcodes' which describe the lectin based on its binding profile. The increased information content encoded by using complex mixtures of a few sugars, rather than increased numbers of different sugars makes this approach both scalable and accessible. These nanoparticles show increased lectin identification power at a range of lectin concentrations, relative to single-channel sensors. It was also found that some information about the concentration of the lectins can be extracted, all from just a simple colour change, taking this technology closer to being a realistic biosensor. PMID:27181289

  5. Glycan heterogeneity on gold nanoparticles increases lectin discrimination capacity in label-free multiplexed bioassays†

    Science.gov (United States)

    Otten, Lucienne; Vlachou, Denise; Richards, Sarah-Jane; Gibson, Matthew I.

    2016-01-01

    The development of new analytical tools as point-of-care biosensors is crucial to combat the spread of infectious diseases, especially in the context of drug-resistant organisms, or to detect biological warfare agents. Glycan/lectin interactions drive a wide range of recognition and signal transduction processes within nature and are often the first site of adhesion/recognition during infection making them appealing targets for biosensors. Glycosylated gold nanoparticles have been developed that change colour from red to blue upon interaction with carbohydrate-binding proteins and may find use as biosensors, but are limited by the inherent promiscuity of some of these interactions. Here we mimic the natural heterogeneity of cell-surface glycans by displaying mixed monolayers of glycans on the surface of gold nanoparticles. These are then used in a multiplexed, label-free bioassay to create ‘barcodes’ which describe the lectin based on its binding profile. The increased information content encoded by using complex mixtures of a few sugars, rather than increased numbers of different sugars makes this approach both scalable and accessible. These nanoparticles show increased lectin identification power at a range of lectin concentrations, relative to single-channel sensors. It was also found that some information about the concentration of the lectins can be extracted, all from just a simple colour change, taking this technology closer to being a realistic biosensor. PMID:27181289

  6. A computational approach for exploring carbohydrate recognition by lectins in innate immunity

    Directory of Open Access Journals (Sweden)

    Mark eAgostino

    2011-06-01

    Full Text Available Recognition of pathogen-associated carbohydrates by a broad range of carbohydrate binding proteins is central to both adaptive and innate immunity. A large functionally diverse group of mammalian carbohydrate binding proteins are lectins, which often display calcium-dependent carbohydrate interactions mediated by one or more carbohydrate recognition domains. We report here the application of molecular docking and site mapping to study carbohydrate recognition by several lectins involved in innate immunity or in modulating adaptive immune responses. It was found that molecular docking programs can identify the correct carbohydrate binding mode, but often have difficulty in ranking it as the best pose. This is largely attributed to the broad and shallow nature of lectin binding sites, and the high flexibility of carbohydrates. Site mapping is very effective at identifying lectin residues involved in carbohydrate recognition, especially with cases that were found to be particularly difficult to characterize via molecular docking. This study highlights the need for alternative strategies to examine carbohydrate-lectin interactions, and specifically demonstrates the potential for mapping methods to extract additional and relevant information from the ensembles of binding poses generated by molecular docking.

  7. The biofilm matrix of Campylobacter jejuni determined by fluorescence lectin-binding analysis.

    Science.gov (United States)

    Turonova, Hana; Neu, Thomas R; Ulbrich, Pavel; Pazlarova, Jarmila; Tresse, Odile

    2016-05-01

    Campylobacter jejuni is responsible for the most common bacterial foodborne gastroenteritis. Despite its fastidious growth, it can survive harsh conditions through biofilm formation. In this work, fluorescence lectin-binding analysis was used to determine the glycoconjugates present in the biofilm matrix of two well-described strains. Screening of 72 lectins revealed strain-specific patterns with six lectins interacting with the biofilm matrix of both strains. The most common sugar moiety contained galactose and N-acetylgalactosamine. Several lectins interacted with N-acetylglucosamine and sialic acid, probably originated from the capsular polysaccharides, lipooligosaccharides and N-glycans of C. jejuni. In addition, glycoconjugates containing mannose and fucose were detected within the biofilm, which have not previously been found in the C. jejuni envelope. Detection of thioflavin T and curcumin highlighted the presence of amyloids in the cell envelope without association with specific cell appendages. The lectins ECA, GS-I, HMA and LEA constitute a reliable cocktail to detect the biofilm matrix of C. jejuni. PMID:27097059

  8. Purification and molecular cloning of a new galactose-specific lectin from Bauhinia variegata seeds

    Indian Academy of Sciences (India)

    Luciano S Pinto; Celso S Nagano; Taianá M Oliveira; Tales R Moura; Alexandre H Sampaio; Henri Debray; Vicente P Pinto; Odir A Dellagostin; Benildo S Cavada

    2008-09-01

    A new galactose-specific lectin was purified from seeds of a Caesalpinoideae plant, Bauhinia variegata, by affinity chromatography on lactose–agarose. Protein extracts haemagglutinated rabbit and human erythrocytes (native and treated with proteolytic enzymes), showing preference for rabbit blood treated with papain and trypsin. Among various carbohydrates tested, the lectin was best inhibited by D-galactose and its derivatives, especially lactose. SDS-PAGE showed that the lectin, named BVL, has a pattern similar to other lectins isolated from the same genus, Bauhinia purpurea agglutinin (BPA). The molecular mass of BVL subunit is 32 871 Da, determined by MALDI-TOF spectrometry. DNA extracted from B. variegata young leaves and primers designed according to the B. purpurea lectin were used to generate specific fragments which were cloned and sequenced, revealing two distinct isoforms. The bvl gene sequence comprised an open reading frame of 876 base pairs which encodes a protein of 291 amino acids. The protein carried a putative signal peptide. The mature protein was predicted to have 263 amino acid residues and 28 963 Da in size.

  9. A comparative study of lectin affinity based plant n-glycoproteome profiling using tomato fruit as a model

    Science.gov (United States)

    Lectin affinity chromatography (LAC) can provide a valuable front-end enrichment strategy for the study of N-glycoproteins and has been used to characterize a broad range eukaryotic N-glycoproteomes. Moreover, studies with mammalian systems have suggested that the use of multiple lectins with differ...

  10. Differential lectin labelling of circulating hemocytes from Biomphalaria glabrata and Biomphalaria tenagophila resistant or susceptible to Schistosoma mansoni infection

    Directory of Open Access Journals (Sweden)

    RL Martins-Souza

    2006-10-01

    Full Text Available Lectins/carbohydrate binding can be involved in the Schistosoma mansoni recognition and activation of the Biomphalaria hemocytes. Therefore, expression of lectin ligands on Biomphalaria hemocytes would be associated with snail resistance against S. mansoni infection. To test this hypothesis, circulating hemocytes were isolated from B. glabrata BH (snail strain highy susceptible to S. mansoni, B. tenagophila Cabo Frio (moderate susceptibility, and B. tenagophila Taim (completely resistant strains, labelled with FITC conjugated lectins (ConA, PNA, SBA, and WGA and analyzed under fluorescence microscopy. The results demonstrated that although lectin-labelled hemocytes were detected in hemolymph of all snail species tested, circulating hemocytes from both strains of B. tenagophila showed a larger number of lectin-labelled cells than B. glabrata. Moreover, most of circulating hemocytes of B. tenagophila were intensively labelled by lectins PNA-FITC and WGA-FITC, while in B. glabrata small hemocytes were labeled mainly by ConA. Upon S. mansoni infection, lectin-labelled hemocytes almost disappeared from the hemolymph of Taim and accumulated in B. glabrata BH. The role of lectins/carbohydrate binding in resistance of B. tengophila infection to S. mansoni is still not fully understood, but the data suggest that there may be a correlation to its presence with susceptibility or resistance to the parasite.

  11. Immunotoxicological studies of genetically modified rice expressing PHA-E lectin or Bt toxin in Wistar rats

    DEFF Research Database (Denmark)

    Kroghsbo, Stine; Madsen, Charlotte Bernhard; Poulsen, Morten;

    2008-01-01

    As part of the SAFOTEST project the immunmodulating effect of Cry1Ab protein from Bacillus thuringiensis (Bt) and PHA-E lectin from kidney bean (Phaseolus vulgaris erythroagglutinin) was examined in 28- and 90-day feeding studies in Wistar rats. PHA-E lectin was chosen as positive control. Rats...

  12. Mannan-binding lectin (MBL) and MBL-associated serine protease 2 (MASP-2) genotypes in colorectal cancer

    DEFF Research Database (Denmark)

    Ytting, H; Christensen, I J; Steffensen, Rudi Nora;

    2011-01-01

    Mannan-binding lectin (MBL) and MBL-associated serine protease 2 (MASP-2) are key factors of the lectin pathway of complement activation. Polymorphisms of the MBL2 and MASP-2 genes affect serum levels of MBL and MASP-2. In patients with colorectal cancer (CRC), the MBL and MASP-2 serum levels are...

  13. Glycomic profiling of developmental changes in bovine testis by lectin histochemistry and further analysis of the most prominent alteration on the level of the glycoproteome by lectin blotting and lectin affinity chromatography

    OpenAIRE

    J. C. Manning; Seyrek, K.; Kaltner, H.; André, S; Sinowatz, F; Gabius, H.J.

    2004-01-01

    The emerging concept of the sugar code attributes functional significance to oligosaccharides of cellular glycoconjugates by protein (lectin)-carbohydrate interactions. Hence it follows that monitoring of glycan expression (glycomic profiling) is not only valuable to delineate characteristic (phenomenological) changes in the cell’s glycosylation but will also come up with the localization of epitopes with potential in biorecognition. It is for this purpose that...

  14. Lectin binding studies on a glycopolymer brush flow-through biosensor by localized surface plasmon resonance.

    Science.gov (United States)

    Rosencrantz, Ruben R; Nguyen, Vu Hoa; Park, Hyunji; Schulte, Christine; Böker, Alexander; Schnakenberg, Uwe; Elling, Lothar

    2016-08-01

    A localized surface plasmon resonance biosensor in a flow-through configuration was applied for investigating kinetics of lectin binding to surface-grafted glycopolymer brushes. Polycarbonate filter membranes with pore sizes of 400 nm were coated with a 114-nm thick gold layer and used as substrate for surface-initiated atom-transfer radical polymerization of a glycomonomer. These grafted from glycopolymer brushes were further modified with two subsequent enzymatic reactions on the surface to yield an immobilized trisaccharide presenting brush. Specific binding of lectins including Clostridium difficile toxin A receptor domain to the glycopolymer brush surface could be investigated in a microfluidic setup with flow-through of the analytes and transmission surface plasmon resonance spectroscopy. Graphical abstract Glycopolymer brushes serve as high affinity ligands for lectin and toxin interactions in a sensitive, disposable flow-through LSPR biosensor. PMID:27277814

  15. The pattern-recognition molecule mannan-binding lectin (MBL) in the pathophysiology of diabetic nephropathy

    DEFF Research Database (Denmark)

    Axelgaard, Esben; Thiel, Steffen; Hansen, Troels Krarup;

    The pattern-recognition molecule mannan-binding lectin (MBL) in the pathophysiology of diabetic nephropathy Esben Axelgaard*; Steffen Thiel*; Jakob Appel Østergaard† and Troels Krarup Hansen† *Department of Biomedicine, Aarhus University, Wilhelm Meyer´s Allé 4, 8000 Aarhus C, Denmark. †Department...... pattern-recognition molecules that may initiate the lectin pathway of complement activation. As opposed to obvious evolutionary beneficial effects of the complement system, MBL is associated with adverse effects in several diseases including diabetes. There is evidence that link MBL to poor prognosis for...... poorly elucidated. Two putative mechanisms are proposed, 1) the formation of neoepitopes for MBL pattern recognition on host cells would enable lectin pathway activation and 2) inactivation of complement regulatory proteins by glycation that may exaggerate complement attack on host cells. MBL initiates...

  16. Soybean Lectin Enhances Biofilm Formation by Bradyrhizobium japonicum in the Absence of Plants

    Directory of Open Access Journals (Sweden)

    Julieta Pérez-Giménez

    2009-01-01

    Full Text Available Soybean lectin (SBL purified from soybean seeds by affinity chromatography strongly bound to Bradyrhizobium japonicum USDA 110 cell surface. This lectin enhanced biofilm formation by B. japonicum in a concentration-dependent manner. Presence of galactose during biofilm formation had different effects in the presence or absence of SBL. Biofilms were completely inhibited in the presence of both SBL and galactose, while in the absence of SBL, galactose was less inhibitory. SBL was very stable, since its agglutinating activity of B. japonicum cells as well as of human group A+ erythrocytes was resistant to preincubation for one week at 60°C. Hence, we propose that plant remnants might constitute a source of this lectin, which might remain active in soil and thus favor B. japonicum biofilm formation in the interval between soybean crop seasons.

  17. The typing of Staphylococcus epidermidis by a lectin-binding assay

    DEFF Research Database (Denmark)

    Jarløv, J O; Hansen, J E; Rosdahl, V T;

    1992-01-01

    A new typing method for Staphylococcus epidermidis was developed. Four biotinylated lectins--wheat germ agglutinin (WGA), soy bean agglutinin (SBA), lentil agglutinin (LCA) and Concanavalin A (ConA)--were added to immobilised whole cells of coagulase-negative staphylococci (CNS) in microtitration...... plates. The amount of bound lectin was measured by peroxidase-conjugated avidin followed by a peroxidase reaction. The method was compared to antibiotic-resistance analysis, phage typing, plasmid DNA profiles and slime production. A total of 113 isolates of CNS from 21 patients was investigated and 71...... strains of CNS, including 64 strains of S. epidermidis, were detected if all typing methods were taken into consideration. If only one typing method was used the highest discriminatory power among the S. epidermidis isolates was obtained with the lectin-binding assay which allowed 49 different strains to...

  18. Evaluation of Parkia pendula lectin mRNA differentially expressed in seedlings.

    Science.gov (United States)

    Rêgo, M J B M; Santos, P B; Carvalho-Junior, L B; Stirling, J; Beltrão, E I C

    2014-05-01

    Parkia pendula (Willd.) Walp. (Fabaceae) is a neotropical species of the genus Parkia more abundantly distributed in Central to South America. From the seeds of P. pendula a glucose/mannose specific lectin (PpeL) was isolated that has been characterised and used as a biotechnological tool but until now this is the first manuscript to analyse P. pendula mRNA expression in seedlings. For this porpoise a Differential display reverse transcription polimerase chain reaction (DDRT-PCR) was used to evaluate the expression of P. pendula lectin mRNAs in non-rooted seedlings. No bands were observed in the agarose gel, indicating the absence of mRNA of PpeL seedlings. our findings confirm that lectins mRNAs are differently regulated among species even if they are grouped in the same class. PMID:25166336

  19. Morphological Specifications of the Bird Schistosome Cercariae and Surface Carbohydrates as Receptors for Lectins

    Directory of Open Access Journals (Sweden)

    I Moebedi

    2007-04-01

    Full Text Available Background: To determine the morphological specifications of the bird schistosomes cercaria from Lymnaea gedrosiana and to detect the surface carbohydrates as receptors for host lectins in the host-parasite relationship systems such as avian schistosomiasis and human cercarial dermatitis. Methods: One hundred ninety two snails collected from Dezful areas in Khuzestan Province, in the south west of Iran, during 2005-2006 were examined for cercariae using shedding and crushing methods. In addition, surface carbohydrates on the cercariae were detected by lentil (Lens culinaris lectins. Results: From the total number of Lymnaea gedrosiana, which examined for bird schistosomes cercaria, 9(4% snails were found to be infected with furcocercus cercaria of the bird schistosomes (probably Gigantobilharzia sp.. Mannose monosaccharide CH2OH (CHOH4CHO as surface carbohydrate was also detected on the cercariae. Conclusion: Mannose carbohydrate on these cercariae may be used as receptor by lectins.

  20. Crystallization and preliminary X-ray diffraction analysis of the lectin from Dioclea rostrata Benth seeds

    International Nuclear Information System (INIS)

    D. rostrata lectin was crystallized by hanging-drop vapor diffusion. The crystal belongs to the orthorhombic space group I222 and diffracted to 1.87 Å resolution. Lectins from the Diocleinae subtribe (Leguminosae) are highly similar proteins that promote various biological activities with distinctly differing potencies. The structural basis for this experimental data is not yet fully understood. Dioclea rostrata lectin was purified and crystallized by hanging-drop vapour diffusion at 293 K. The crystal belongs to the orthorhombic space group I222, with unit-cell parameters a = 61.51, b = 88.22, c = 87.76 Å. Assuming the presence of one monomer per asymmetric unit, the solvent content was estimated to be about 47.9%. A complete data set was collected at 1.87 Å resolution

  1. Lectin-microarray technique for glycomic profiling of fungal cell surfaces.

    Science.gov (United States)

    Shibazaki, Azusa; Gonoi, Tohru

    2014-01-01

    Lectin microarrays are rows of lectins with different carbohydrate-binding specificities spotted on surfaces of glass slides. Lectin microarray technique enables glycomic analyses of carbohydrate composition of fungal cell walls. We will describe an application of the technique in analyzing cell surface glycome of yeast-form fungal cells in the living state. The analysis reveals genus- and species-dependent complex cell surface carbohydrate structures of fungi, and enabled us, therefore, to suggest that cell walls of yeast cells, which have been considered to have relatively simple structures, actually have a more complex structure containing galactose and fucose. This shows that the technique can be used to find new insights into the study of phylogenetic relations and into the classification of cells in the fungal kingdom based on cell wall glycome. PMID:25117243

  2. Experimental study of the occurence and properties of C-type retroviruses in radiation-induced osteosarcomas in mice

    International Nuclear Information System (INIS)

    In the radiation induced osteosarcomas of the C 3 Hx101/F1-mouse C-type virus particles had been found regularly with a density of 1.16 g/cm3, with high molecular RNA, a reverse transcriptase and the murine group-specific antigen p 30. Osteosarcomas of the NMRI-mouse, however, had only p 30 protein and so-called intra-cisternal A-type particles. After 'in vitro' cultivation retroviruses had been liberated from the osteosarcoma cells of the C 3 Hx101/F1-mice as well as from the NMRI-mice type C. During the tumour latency period a virus expression of the C-type retroviruses had been found for a certain period in the first month after irradiation of the bone tissue had begun; then followed an antibody-reaction which continued to persist until the 8th month. Another virus expression was observed in the skeleton during the period when the osteosarcomas appeared. This virus expression was accompanied by a decrease in antibodies and a temporary increase of the viral p 30 protein in the serum. The viruses which had been isolated from the radiation induced osteosarcomas showed the properties which are typical for ecotropic C-type retroviruses of mice. After infection of new-born mice these viruses produced fibrosarcomas (C 3 Hx 101/F1-mice) or lymphomas and osteomas (NMRI-mice). The results make it obvious that the endogenetic C-type retroviruses participate in the formation of radiation-induced sarcomas in mice. (orig./MG)

  3. Four functionally distinct C-type natriuretic peptides found in fish reveal evolutionary history of the natriuretic peptide system

    OpenAIRE

    Inoue, Koji; Naruse, Kiyoshi; Yamagami, Sayaka; Mitani, Hiroshi; Suzuki, Norio; Takei, Yoshio

    2003-01-01

    Natriuretic peptides (NPs) are major cardiovascular and osmoregulatory hormones in vertebrates. Although tetrapods generally have three subtypes, atrial NP (ANP), B-type NP (BNP), and C-type NP (CNP), some teleosts lack BNP, and sharks and hagfish have only one NP. Thus, NPs have diverged during fish evolution, possibly reflecting changes in osmoregulatory systems. In this study, we found, by cDNA cloning, four distinct CNPs (1 through 4) in the medaka (Oryzias latipes...

  4. Flow cytometric analysis of lectin binding to in vitro-cultured Perkinsus marinus surface carbohydrates

    Science.gov (United States)

    Gauthier, J.D.; Jenkins, J.A.; La Peyre, Jerome F.

    2004-01-01

    Parasite surface glycoconjugates are frequently involved in cellular recognition and colonization of the host. This study reports on the identification of Perkinsus marinus surface carbohydrates by flow cytometric analyses of fluorescein isothiocyanate-conjugated lectin binding. Lectin-binding specificity was confirmed by sugar inhibition and Kolmogorov-Smirnov statistics. Clear, measurable fluorescence peaks were discriminated, and no parasite autofluorescence was observed. Parasites (GTLA-5 and Perkinsus-1 strains) harvested during log and stationary phases of growth in a protein-free medium reacted strongly with concanavalin A and wheat germ agglutinin, which bind to glucose-mannose and N-acetyl-D-glucosamine (GlcNAc) moieties, respectively. Both P. marinus strains bound with lower intensity to Maclura pomifera agglutinin, Bauhinia purpurea agglutinin, soybean agglutinin (N-acetyl-D-galactosamine-specific lectins), peanut agglutinin (PNA) (terminal galactose specific), and Griffonia simplicifolia II (GlcNAc specific). Only background fluorescence levels were detected with Ulex europaeus agglutinin I (L-fucose specific) and Limulus polyphemus agglutinin (sialic acid specific). The lectin-binding profiles were similar for the 2 strains except for a greater relative binding intensity of PNA for Perkinsus-1 and an overall greater lectin-binding capacity of Perkinsus-1 compared with GTLA-5. Growth stage comparisons revealed increased lectin-binding intensities during stationary phase compared with log phase of growth. This is the first report of the identification of surface glycoconjugates on a Perkinsus spp. by flow cytometry and the first to demonstrate that differential surface sugar expression is growth phase and strain dependent. ?? American Society of Parasitologists 2004.

  5. 植物凝集素的功能%Plant lectins: multifunctions and applications

    Institute of Scientific and Technical Information of China (English)

    鲍锦库

    2011-01-01

    Plant lectins are carbohydrate-binding proteins with nonimmune origin, and they could bind carbohydrates reversibly, agglutinate cells or precipitate polysaccharides and glycoconjugates. Due to the specific binding activities towards polysaccharides and glycoconjugates of plant lectins, they play important roles in signaling pathways, immunoregulation and plant defence. In addition, plant lectins also possess hemagglutinating and carbohydrate-binding activities as well as the anti-viral and apoptosis-inducing properties. Therefore, plant lectins have been drawn a rising attention for researchers in bioscience, medicine and agriculture. In this review, we summarize the history and multifunctions of plant lectins, and also present the applications of plant lectins.%植物凝集素是来源于植物的一类能凝集细胞和沉淀单糖或多糖复合物的非免疫来源的非酶蛋白质.由于其对于单糖或糖复合物特异性结合的能力,使得其在如信号转导、免疫反应、植物防御等诸多信号过程中均具有重要作用.同时植物凝集素具有细胞凝集、抗病毒、抗真菌及诱导细胞凋亡或自噬等多种能力,因此在生命科学、医学及农业方面均有较好的研究价值和应用前景.综述了植物凝集素的研究历史和凝集素的主要功能,并对现阶段凝集素的重点应用做简要介绍.

  6. Reactivities of N-acetylgalactosamine-specific lectins with human IgA1 proteins.

    Science.gov (United States)

    Moore, Jennifer S; Kulhavy, Rose; Tomana, Milan; Moldoveanu, Zina; Suzuki, Hitoshi; Brown, Rhubell; Hall, Stacy; Kilian, Mogens; Poulsen, Knud; Mestecky, Jiri; Julian, Bruce A; Novak, Jan

    2007-04-01

    Lectins are proteins with specificity of binding to certain monosaccharides or oligosaccharides. They can detect abnormal glycosylation patterns on immunoglobulins in patients with various chronic inflammatory diseases, including rheumatoid arthritis and IgA nephropathy (IgAN). However, lectins exhibit binding heterogeneity, depending on their source and methods of isolation. To characterize potential differences in recognition of terminal N-acetylgalactosamine (GalNAc) on IgA1, we evaluated the binding characteristics of several commercial preparations of GalNAc-specific lectins using a panel of IgA1 and, as controls, IgA2 and IgG myeloma proteins. These lectins originated from snails Helix aspersa (HAA) and Helix pomatia (HPA), and the plant Vicia villosa (VV). Only HAA and HPA bound exclusively to IgA1, with its O-linked glycans composed of GalNAc, galactose, and sialic acid. In contrast, VV reacted with sugars of both IgA subclasses and IgG, indicating that it also recognized N-linked glycans without GalNAc. Furthermore, HAA and HPA from several manufacturers differed in their ability to bind various IgA1 myeloma proteins and other GalNAc-containing glycoproteins in ELISA and Western blot. For serum samples from IgAN patients, HAA was the optimal lectin to study IgA1 glycosylation in ELISA and Western blot assays, including identification of the sites of attachment of the aberrant glycans. The galactose-deficient glycans were site-specific, localized mostly at Thr228 and/or Ser230. Because of the heterogeneity of GalNAc-specific lectins, they should be carefully characterized with appropriate substrates before undertaking any study. PMID:17275907

  7. Lectin histochemistry of intestinal carbohydrate determinants in representatives of different classes of vertebrates

    Directory of Open Access Journals (Sweden)

    Antonyuk R.V.

    2015-12-01

    Full Text Available Background. Glycoproteins (including mucin of vertebrate’s intestine play an important role in its protection against chemical and mechanical damage and bacterial attacks. Their diversity was described by many authors, but understanding of their chemical structure remains far from complete. These data can be extended by methods of lectin histochemistry. Objective. To investigate the rearrangement of intestinal carbohydrate determinants in the context of vertebrate evolution. Methods. Distal and proximal segments of small and large intestines of humans (Homo sapiens, laboratory (Wistar rat (Rattus norvegicus f. Domesticus, rock pigeon (Columba livia, smooth snake (Coronella austriaca, common frog (Rana temporaria, common carp (Cyprinus carpio that belong to different classes of vertebrates were taken for the experiment. Nine lectins with different carbohydrate specificities: wheat germ (WGA, potato (STA, elderberry bark (SNA, golden rain bark (LABA, locust bark (RPBA, roe carp (CCRA, Phaseolus vulgaris erytroagglutinin (PHA-E, peanut (PNA and jack fruit (AIA – were included into the panel. Results. Differences in lectin staining between small and large intestine were more pronounced in higher (human, rat than in lower (frog, carp vertebrates. Lectin receptors were more diverse in frog intestine in comparison with carp. Lectin interaction with mucin secretory granules of smooth snake revealed lack of N-acetyl-D-glucosamine residues and abundance of N-acetyl-D-galactosamine determinants. Conclusion. Intestines of all studied vertebrate species demonstrate high content of secretory mucins that exposed terminal acidic carbohydrates including sialic acid. The diversity and differences in the structure of glycans of the digestive tract of vertebrates is apparently determined by several factors – diet, environmental and living conditions, intestinal microbiota interactions etc. Citation: Antonyuk RV, Lutsyk AD. [Lectin histochemistry of intestinal

  8. New crystal forms of Diocleinae lectins in the presence of different dimannosides

    International Nuclear Information System (INIS)

    The crystallization and preliminary X-ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(α1-2)Man(α1)OMe, Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe in two crystal forms [the complexes with Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe crystallized in space group P32 and those with Man(α1-2)Man(α1)OMe crystallized in space group I222], which differed from those of the native proteins (P21212 for CML and C222 for CGL), are reported. Studying the interactions between lectins and sugars is important in order to explain the differences observed in the biological activities presented by the highly similar proteins of the Diocleinae subtribe. Here, the crystallization and preliminary X-ray data of Canavalia gladiata lectin (CGL) and C. maritima lectin (CML) complexed with Man(α1-2)Man(α1)OMe, Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe in two crystal forms [the complexes with Man(α1-3)Man(α1)OMe and Man(α1-4)Man(α1)OMe crystallized in space group P32 and those with Man(α1-2)Man(α1)OMe crystallized in space group I222], which differed from those of the native proteins (P21212 for CML and C222 for CGL), are reported. The crystal complexes of ConA-like lectins with Man(α1-4)Man(α1)OMe are reported here for the first time

  9. Carbohydrate recognition by the antiviral lectin cyanovirin-N.

    Science.gov (United States)

    Fujimoto, Yukiji K; Green, David F

    2012-12-01

    Cyanovirin-N (CVN) is a cyanobacterial lectin with potent antiviral activity and has been the focus of extensive preclinical investigation as a potential prophylactic for the prevention of the sexual transmission of the human immunodeficiency virus (HIV). Here we present a detailed analysis of carbohydrate recognition by this important protein, using a combination of computational methods, including extensive molecular dynamics simulations and molecular mechanics/Poisson-Boltzmann surface area (MM/PBSA) energetic analysis. The simulation results strongly suggest that the observed tendency of wild-type CVN to form domain-swapped dimers is the result of a previously unidentified cis-peptide bond present in the monomeric state. The energetic analysis additionally indicates that the highest-affinity ligand for CVN characterized to date (α-Man-(1,2)-α-Man-(1,2)-α-Man) is recognized asymmetrically by the two binding sites. Finally, we are able to provide a detailed map of the role of all binding site functional groups (both backbone and side chain) to various aspects of molecular recognition: general affinity for cognate ligands, specificity for distinct oligosaccharide targets, and the asymmetric recognition of α-Man-(1,2)-α-Man-(1,2)-α-Man. Taken as a whole, these results complement past experimental characterization (both structural and thermodynamic) to provide the most complete understanding of carbohydrate recognition by CVN to date. The results also provide strong support for the application of similar approaches to the understanding of other protein-carbohydrate complexes. PMID:23057413

  10. Evaluation of ocular surface glycocalyx using lectin-conjugated fluorescein

    Directory of Open Access Journals (Sweden)

    Hiroshi Mochizuki

    2010-08-01

    Full Text Available Hiroshi Mochizuki1, Masaki Fukui1,2, Shin Hatou1,2, Masakazu Yamada1, Kazuo Tsubota21Division for Vision Research, National Institute of Sensory Organs, National Tokyo Medical Center, Tokyo, Japan; 2Department of Ophthalmology, Keio University School of Medicine, Tokyo, JapanPurpose: A glycocalyx plays important roles in the ocular surface epithelium, but there is no direct simple method to evaluate ocular surface glycocalyx. We tested a wheat germ agglutinin conjugate of fluorescein (F-WGA as a marker to demonstrate ocular surface glycocalyx in vivo.Methods: After a 5% F-WGA solution was applied to the eyes of eight healthy volunteers, fluorescent intensities of the central cornea and bulbar conjunctiva were measured by fluorophotometry. A 10% fluorescein-conjugated dextran solution served as the control. Changes in fluorescent intensities of the ocular surface following a challenge with 5% N-acetyl cysteine, a conventional mucolytic agent, were tested in the second experiment. Saline instillation served as a control.Results: The ocular surface was diffusely stained by F-WGA. Breakup of the precorneal tear film was not apparent, possibly because F-WGA was bound to the glycocalyx of the ocular surface epithelium. F-WGA fluorescent intensities were high in the superior, nasal, and inferior regions of the bulbar conjunctiva and low in the temporal conjunctiva and cornea. No such regional differences were observed with fluorescein-conjugated dextran. F-WGA fluorescent intensities decreased significantly with N-acetyl cysteine instillation, whereas they were not affected by saline instillation.Conclusion: The fluorophotometric method may be used to evaluate the glycocalyx quantitatively in the ocular surface in vivo.Keywords: cornea, fluorescein, glycocalyx, lectin, mucin

  11. The utility of three dimensional DSA with bi-C-typed arms in the embolization of intracranial aneurysms

    International Nuclear Information System (INIS)

    Objective: To discuss the value of three dimensional DSA(3D-DSA) with bi-C-typed arms on the embolization of intracranial aneurysms. Methods: 3D-DSA was performed in eight patients (aged 43-73 years) with spontaneous subarachnoid hemorrhage (SAH) by frontal, lateral and various oblique projections including rotational angiography and three dimensional reconstruction. The perfect vision angle of aneurysms could be found on the 3D-workstation by the auto positioning function of 3D-DSA which could send the parameters to the main machine and make the bi-C-typed arms turning to the proper angle to have aneurysms embolized totally. Results: Eight SAH cases were found with aneurysms including one micro-aneurysm and four with bleb (6 blebs). Six cases were under-taken selective embolization with 5 of total occlusion showing no bleb in follow up angiography and the other one of partial occlusion. The patient with micro-aneurysm didn't receive embolization and another one received microneurosurgery operation. No blebs were found in the follow up angiography. Conclusions: 3D-DSA with bi-C-typed arms can facilitate in finding intracranial aneurysms, micro-aneurysms and blebs' and provide the best procedural angulation for embolization with more safe. (authors)

  12. Lectin binding profiles of SSEA-4 enriched, pluripotent human embryonic stem cell surfaces

    Directory of Open Access Journals (Sweden)

    Shin Soojung

    2005-07-01

    Full Text Available Abstract Background Pluripotent human embryonic stem cells (hESCs have the potential to form every cell type in the body. These cells must be appropriately characterized prior to differentiation studies or when defining characteristics of the pluripotent state. Some developmentally regulated cell surface antigens identified by monoclonal antibodies in a variety of species and stem cell types have proven to be side chains of membrane glycolipids and glycoproteins. Therefore, to examine hESC surfaces for other potential pluripotent markers, we used a panel of 14 lectins, which were chosen based on their specificity for a variety of carbohydrates and carbohydrate linkages, along with stage specific embryonic antigen-4 (SSEA-4, to determine binding quantitation by flow cytometry and binding localization in adherent colonies by immunocytochemistry. Results Enriching cells for SSEA-4 expression increased the percentage of SSEA-4 positive cells to 98–99%. Using enriched high SSEA-4-expressing hESCs, we then analyzed the binding percentages of selected lectins and found a large variation in binding percentages ranging from 4% to 99% binding. Lycopersicon (tomatoesculetum lectin (TL, Ricinus communis agglutinin (RCA, and Concanavalin A (Con A bound to SSEA-4 positive regions of hESCs and with similar binding percentages as SSEA-4. In contrast, we found Dolichos biflorus agglutinin (DBA and Lotus tetragonolobus lectin (LTL did not bind to hESCs while Phaseolus vulgaris leuco-agglutinin (PHA-L, Vicia villosa agglutinin (VVA, Ulex europaeus agglutinin (UEA, Phaseolus vulgaris erythro-agglutinin (PHA-E, and Maackia amurensis agglutinin (MAA bound partially to hESCs. These binding percentages correlated well with immunocytochemistry results. Conclusion Our results provide information about types of carbohydrates and carbohydrate linkages found on pluripotent hESC surfaces. We propose that TL, RCA and Con A may be used as markers that are associated with the

  13. Inactivation and fragmentation of lectin from Bothrops leucurus snake venom by gamma irradiation

    Science.gov (United States)

    Nunes, E. S.; Souza, M. A. A.; Vaz, A. F. M.; Coelho, L. C. B. B.; Aguiar, J. S.; Silva, T. G.; Guarnieri, M. C.; Melo, A. M. M. A.; Oliva, M. L. V.; Correia, M. T. S.

    2012-04-01

    Gamma radiation alters the molecular structure of biomolecules and is able to mitigate the action of snake venoms and their isolated toxins. The effect of γ-radiation on the folding of Bothrops lecurus venom lectin was measured by a hemagglutinating assay, intrinsic and bis-ANS fluorescence. Intrinsic and bis-ANS fluorescence analyses indicated that irradiation caused unfolding followed by aggregation of the lectin. Our results suggest that irradiation can lead to significant changes in the protein structure, which may promote the loss of its binding property and toxic action.

  14. Sequence of a cysteine-rich galactose-specific lectin of Entamoeba histolytica.

    OpenAIRE

    Mann, B J; Torian, B E; Vedvick, T S; Petri, W A

    1991-01-01

    Entamoeba histolytica trophozoites adhere to human colonic mucins and epithelial cells by a cell surface galactose-specific lectin. This lectin, which is composed of two subunits linked by disulfide bonds, has been shown to be a protective antigen in an animal model of amebiasis. We have determined the sequence of the mature form of the 170-kDa heavy subunit from cDNA clones and PCR-amplified fragments. The heavy subunit sequence consisted of a putative extracellular domain containing 1209 am...

  15. Evaluation of Fluorescently Labeled Lectins for Noninvasive Localization of Extracellular Polymeric Substances in Sphingomonas Biofilms

    OpenAIRE

    Johnsen, Anders R.; Hausner, Martina; Schnell, Annette; Wuertz, Stefan

    2000-01-01

    Three strains of Sphingomonas were grown as biofilms and tested for binding of five fluorescently labeled lectins (Con A-type IV-TRITC or -Cy5, Pha-E-TRITC, PNA-TRITC, UEA 1-TRITC, and WGA-Texas red). Only ConA and WGA were significantly bound by the biofilms. Binding of the five lectins to artificial biofilms made of the commercially available Sphingomonas extracellular polysaccharides was similar to binding to living biofilms. Staining of the living and artificial biofilms by ConA might be ...

  16. Identification of endogenous binding proteins for the lectin discoidin-I in Dictyostelium discoideum.

    OpenAIRE

    W. Breuer; Siu, C H

    1981-01-01

    Recent biochemical and genetic evidence has shown that the endogenous lectin discoidin-I is involved in intercellular adhesion during development of the cellular slime mold Dictyostelium discoideum. We have prepared discoidin-I affinity columns and used them to isolate the lectin receptors. By using the cell surface radioiodination method, 11 discoidin-I binding proteins were identified in wild-type NC4 cells by gel electrophoresis, with apparent molecular weights of 95,000, 85,000, 78,000, 7...

  17. Epidemiology of chronic wound patients and relation to serum levels of mannan-binding lectin

    DEFF Research Database (Denmark)

    Bitsch, Mikael; Laursen, Inga; Engel, Anne-Mari;

    2009-01-01

    consecutive patients with chronic foot and leg ulcers. A clinical classification of wound- aetiology was performed, and mannan-binding lectin was measured in the sera of patients and healthy controls. The patients presented with 639 wounds altogether; diabetic foot ulcers (309), venous leg ulcers (188......), arterial ulcers (109), and vasculitis (33). The mannan-binding lectin levels of patients with venous leg ulcer, alone or in combination with other types of wounds, differed significantly from the control group, and the frequency of values < 100 ng/ml was significantly higher. In diabetic and arterial ulcer...

  18. Inhibition of the adenylylation of liver plasma membrane-bound proteins by plant and mammalian lectins

    OpenAIRE

    San José, Esteban; Villalobo, Eduarde; Gabius, Hans-J.; Villalobo, Antonio

    1993-01-01

    Liver plasma membrane contains four major (130-kDa, 120-kDa, 110-kDa and 100-kDa) sialic acid-containing glycopolypeptides that are able to undergo adenylylation, as well as phosphorylation (San José et al. (1990) J. Biol. Chem. 265; 20653-20661). To gain insight into the regulation of these processes, lectins are employed to probe the extent of influence of their interaction with membrane fractions for these reactions. We demonstrate that the beta-galactoside-specific lectins from bovine hea...

  19. Plumieribetin, a Fish Lectin Homologous to Mannose-binding B-type Lectins, Inhibits the Collagen-binding α1β1 Integrin*

    OpenAIRE

    de Santana Evangelista, Karla; Andrich, Filipe; Figueiredo de Rezende, Flávia; Niland, Stephan; Cordeiro, Marta N.; Horlacher, Tim; Castelli, Riccardo; Schmidt-Hederich, Alletta; Peter H. Seeberger; Sanchez, Eladio F.; Richardson, Michael; Gomes de Figueiredo, Suely; Eble, Johannes A.

    2009-01-01

    Recently, a few fish proteins have been described with a high homology to B-type lectins of monocotyledonous plants. Because of their mannose binding activity, they have been ascribed a role in innate immunity. By screening various fish venoms for their integrin inhibitory activity, we isolated a homologous protein from the fin stings and skin mucus of the scorpionfish (Scorpaena plumieri). This protein inhibits α1β1 integrin binding to basement membrane collagen IV. By protein chemical and s...

  20. Mouse Ficolin B Has an Ability to Form Complexes with Mannose-Binding Lectin-Associated Serine Proteases and Activate Complement through the Lectin Pathway

    Directory of Open Access Journals (Sweden)

    Yuichi Endo

    2012-01-01

    Full Text Available Ficolins are thought to be pathogen-associated-molecular-pattern-(PAMP- recognition molecules that function to support innate immunity. Like mannose-binding lectins (MBLs, most mammalian ficolins form complexes with MBL-associated serine proteases (MASPs, leading to complement activation via the lectin pathway. However, the ability of murine ficolin B, a homologue of human M-ficolin, to perform this function is still controversial. The results of the present study show that ficolin B in mouse bone marrow is an oligomeric protein. Ficolin B, pulled down using GlcNAc-agarose, contained very low, but detectable, amounts of MASP-2 and small MBL-associated protein (sMAP and showed detectable C4-deposition activity on immobilized N-acetylglucosamine. These biochemical features of ficolin B were confirmed using recombinant mouse ficolin B produced in CHO cells. Taken together, these results suggest that like other mammalian homologues, murine ficolin B has an ability to exert its function via the lectin pathway.

  1. Use of Aleuria alantia Lectin Affinity Chromatography to Enrich Candidate Biomarkers from the Urine of Patients with Bladder Cancer

    Directory of Open Access Journals (Sweden)

    Sarah R. Ambrose

    2015-09-01

    Full Text Available Developing a urine test to detect bladder tumours with high sensitivity and specificity is a key goal in bladder cancer research. We hypothesised that bladder cancer-specific glycoproteins might fulfill this role. Lectin-ELISAs were used to study the binding of 25 lectins to 10 bladder cell lines and serum and urine from bladder cancer patients and non-cancer controls. Selected lectins were then used to enrich glycoproteins from the urine of bladder cancer patients and control subjects for analysis by shotgun proteomics. None of the lectins showed a strong preference for bladder cancer cell lines over normal urothlelial cell lines or for urinary glycans from bladder cancer patients over those from non-cancer controls. However, several lectins showed a strong preference for bladder cell line glycans over serum glycans and are potentially useful for enriching glycoproteins originating from the urothelium in urine. Aleuria alantia lectin affinity chromatography and shotgun proteomics identified mucin-1 and golgi apparatus protein 1 as proteins warranting further investigation as urinary biomarkers for low-grade bladder cancer. Glycosylation changes in bladder cancer are not reliably detected by measuring lectin binding to unfractionated proteomes, but it is possible that more specific reagents and/or a focus on individual proteins may produce clinically useful biomarkers.

  2. Red kidney bean (Phaseolus vulgaris lectin stimulation increases the number of enterochromaffin cells in the small intestine of suckling piglets

    Directory of Open Access Journals (Sweden)

    Zacharko-Siembida Anna

    2014-06-01

    Full Text Available The quantities and distribution patterns of serotonin-immunoreactive (serotonin-IR enterochromaffin cells (EC were studied immunohistochemically in the small intestine of suckling piglets stimulated with red kidney bean lectin, and in nonstimulated, control animals. The co-expression patterns of serotonin with somatostatin (SOM or corticotropin releasing-factor (CRF were also studied. After the lectin treatment, the increased numbers of EC were noted in the duodenum of experimental animals. Lectin stimulation did not change the proportions of EC in the jejunum and ileum. In the duodenal epithelium of the lectin-stimulated piglets, the vast majority of serotonin-IR EC were distributed at the basis of crypts. After the lectin administration, the proportions of serotonin-IR/SOM-IR EC were statistically similar in all sections of the small intestine. No upregulation of CRF was found in duodenal, jejunal, and ileal EC of lectin-treated animals. The findings demonstrated that red kidney bean lectin increased the serotonin reservoir in the duodenum, and thus may be an effective stimulant of the gut maturation in suckling mammals.

  3. Glycophenotype Evaluation in Cutaneous Tumors Using Lectins Labeled with Acridinium Ester

    Directory of Open Access Journals (Sweden)

    Luiza Rayanna Amorim Lima

    2013-01-01

    Full Text Available Background. Tumor cells show alterations in their glycosylation patterns when compared to normal cells. Lectins can be used to evaluate these glycocode changes. Chemiluminescence assay is an effective technique for quantitative analysis of proteins, nucleic acids, and carbohydrates due to its high sensitivity, specificity, and rapid testing. Objective. To use histochemiluminescence based on lectin conjugated to acridinium ester (AE for the investigation of glycophenotype changes in cutaneous tumors. Methods. Concanavalin A (Con A, Peanut agglutinin (PNA, Ulex europaeus agglutinin-I (UEA-I, and Maackia amurensis agglutinin (MAA were conjugated to acridinium ester. Biopsies of cutaneous tumors and normal skin were incubated with the lectins-AE, and chemiluminescence was quantified and expressed as Relative Light Units (RLU. Results. Actinic keratosis (AK, keratoacanthoma (KA, squamous cell carcinoma (SCC, and basal cell carcinoma (BCC showed lower expression of α-D-glucose/mannose and α-L-fucose residues compared to normal tissue. Cutaneous tumors displayed higher expression of Gal-β(1-3-GalNAc residues than normal tissue. AK and SCC exhibited higher expression of Neu5Ac-α(2,3Gal residues than normal epidermis. KA and BCC showed equivalent RLU values compared to normal tissue. Conclusions. Lectin histochemiluminescence allowed quantitative assessment of the carbohydrate expression in cutaneous tissues, contributing to eliminate the subjectivity of conventional techniques used in the histopathological diagnosis.

  4. The mannan-binding lectin pathway of complement activation: biology and disease association

    DEFF Research Database (Denmark)

    Petersen, Steen Vang; Thiel, S; Jensenius, J C

    2001-01-01

    Mannan-binding lectin (MBL) is a plasma protein found in association with several serine proteases (MASPs) forming the MBL complex. MBL recognises carbohydrate structures arranged in a particular geometry, such as those found on the surface of micro-organisms. When bound to e.g. bacteria the MBL...

  5. The use of lectins as markers for differentiated secretory cells in planarians.

    Science.gov (United States)

    Zayas, Ricardo M; Cebrià, Francesc; Guo, Tingxia; Feng, Junjie; Newmark, Phillip A

    2010-11-01

    Freshwater planarians have reemerged as excellent models to investigate mechanisms underlying regeneration. The introduction of molecular tools has facilitated the study of planarians, but cell- and tissue-specific markers are still needed to examine differentiation of most cell types. Here we report the utility of fluorescent lectin-conjugates to label tissues in the planarian Schmidtea mediterranea. We show that 16 lectin-conjugates stain planarian cells or tissues; 13 primarily label the secretory cells, their cytoplasmic projections, and terminal pores. Thus, we examined regeneration of the secretory system using lectin markers and functionally characterized two genes expressed in the secretory cells: marginal adhesive gland-1 (mag-1) and Smed-reticulocalbin1 (Smed-rcn1). RNAi knockdown of these genes caused a dramatic reduction of secretory cell lectin staining, suggesting a role for mag-1 and Smed-rcn1 in secretory cell differentiation. Our results provide new insights into planarian secretory system regeneration and add new markers for labeling several planarian tissues. PMID:20865784

  6. Preoperative mannan-binding lectin pathway and prognosis in colorectal cancer

    DEFF Research Database (Denmark)

    Ytting, Henriette; Christensen, Ib Jarle; Jensenius, Jens Christian;

    2005-01-01

    PURPOSE: Deficiency of the mannan-binding lectin (MBL) pathway of innate immunity is associated with increased susceptibility to infections. In patients with colorectal cancer (CRC), postoperative infection is associated with poor prognosis. The aim of the present study was to evaluate (1) the...

  7. A `Clicked' Tetrameric Hydroxamic Acid Glycopeptidomimetic Antagonizes Sugar-Lectin Interactions On The Cellular Level

    Science.gov (United States)

    Zhang, Hai-Lin; Zang, Yi; Xie, Juan; Li, Jia; Chen, Guo-Rong; He, Xiao-Peng; Tian, He

    2014-07-01

    A tetrameric N-acetyl galactosaminyl (GalNAc) peptidomimetic was constructed by N-acetylation of repeating proline-based hydroxamic acid units, followed by a convergent `click chemistry' coupling. This novel glycopeptidomimetic was determined to effectively antagonize the interaction between a transmembrane hepatic lectin and GalNAc on the cellular level.

  8. A second form of collagenous lectin from the tunicate, Styela plicata.

    NARCIS (Netherlands)

    Green, P.; Luty, J.F.; Nair, S.; Radford, J.; Raftos, D.

    2006-01-01

    This study characterised a 90 kDa lectin from an invertebrate chordate, the tunicate Styela plicata. One- and two-dimensional electrophoresis showed that the apparent molecular weight of this protein is maintained under both reducing and non-reducing conditions, suggesting that its native form is a

  9. Detection of sugar-lectin interactions by multivalent dendritic sugar functionalized single-walled carbon nanotubes

    Science.gov (United States)

    Vasu, K. S.; Naresh, K.; Bagul, R. S.; Jayaraman, N.; Sood, A. K.

    2012-07-01

    We show that single walled carbon nanotubes (SWNTs) decorated with sugar functionalized poly (propyl ether imine) (PETIM) dendrimer is a very sensitive platform to quantitatively detect carbohydrate recognizing proteins, namely, lectins. The changes in electrical conductivity of SWNT in field effect transistor device due to carbohydrate-protein interactions form the basis of present study. The mannose sugar attached PETIM dendrimers undergo charge-transfer interactions with the SWNTs. The changes in the conductance of the dendritic sugar functionalized SWNT after addition of lectins in varying concentrations were found to follow the Langmuir type isotherm, giving the concanavalin A (Con A)-mannose affinity constant to be 8.5 × 106 M-1. The increase in the device conductance observed after adding 10 nM of Con A is same as after adding 20 μM of a non-specific lectin peanut agglutinin, showing the high specificity of the Con A-mannose interactions. The specificity of sugar-lectin interactions was characterized further by observing significant shifts in Raman modes of the SWNTs.

  10. Mannan-binding lectin and MBL-associated serine protease-2

    DEFF Research Database (Denmark)

    Jorgensen, J.; Ytting, H.; Steffensen, R.M.;

    2008-01-01

    used for detection, evaluation of prognosis, therapy selection and monitoring. The serum proteins of the innate immune system mannan-binding lectin (MBL) and MBL-associated serine protease-2 (MASP-2) are novel biomarkers under validation in CRC. Low preoperative MBL levels are predictive of pneumonia...

  11. Ternary supramolecular quantum-dot network flocculation for selective lectin detection

    NARCIS (Netherlands)

    Oikonomou, Maria; Wang, Junyou; Carvalho, Rui Rijo; Velders, Aldrik H.

    2016-01-01

    We present a versatile, tuneable, and selective nanoparticle-based lectin biosensor, based on flocculation of ternary supramolecular nanoparticle networks (NPN), formed through the sequential binding of three building blocks. The three building blocks are β-cyclodextrin-capped CdTe quantum dots,

  12. Isolation and characterization of a novel lectin with antifungal and antiproliferative activities from Sophora alopecuroides seeds

    Institute of Scientific and Technical Information of China (English)

    Tinging Li; Xiaoli Yin; Dongliang Liu; Xiaojin Ma; Hui Lv; Surong Sun

    2012-01-01

    Sophora alopecuroides lectin (SAL),a novel lectin from the seeds of Sophora alopecuroides,was purified by ionexchange chromatography on diethylaminoethyl (DEAE)-and carboxymethyl (CM)-Sepharose columns,followed by gel filtration on a Sephadex 75 10/300 GL column.SAL was found to be a monomer of 39916.3 Da,as determined by tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance liquid chromato-graphy (HPLC).The N-terminal 10-amino acid sequence of SAL,KPWALSFSFG,resembles those of other legume lectins.SAL exhibits hemagglutinating activity against rabbit erythrocytes at 11.9 μg/ml.Its hemagglutinating activity is stable in the pH range 7-11 and in the temperature range 30-90℃,and is stimulated by Mn2+.The hemagglutinating activity of SAL is most potently inhibited by 50-mM D-galactose.SAL suppresses mycelial growth in Penicillium digitatum and Alternaria alternata;the IC50 of the antifungal activity toward P.digitatum and A.alternata were found to be 3.125 and 3.338 μM,respectively.SAL suppresses the proliferation of human cervical cancer cells (HeLa) at an ICso of 6.25 μM (P< 0.05).But it has no inhibiting effect on bacteria.This is the first report of a lectin from seeds of S.alopecuroides.

  13. Mannose-binding lectin-2 genotypes and recurrent late pregnancy losses

    DEFF Research Database (Denmark)

    Christiansen, Ole B; Nielsen, Henriette S; Lund, Marie;

    2008-01-01

    BACKGROUND: Low levels of mannose-binding lectin (MBL) predispose to various infectious and inflammatory disorders and have been reported to be associated with recurrent early miscarriages. Recurrent late pregnancy losses (RLPL) in the second trimester is a rare but devastating syndrome where...

  14. Isolation and characterization of a novel lectin with antifungal and antiproliferative activities from Sophora alopecuroides seeds.

    Science.gov (United States)

    Li, Tingting; Yin, Xiaoli; Liu, Dongliang; Ma, Xiaojin; Lv, Hui; Sun, Surong

    2012-07-01

    Sophora alopecuroides lectin (SAL), a novel lectin from the seeds of Sophora alopecuroides, was purified by ion-exchange chromatography on diethylaminoethyl (DEAE)- and carboxymethyl (CM)-Sepharose columns, followed by gel filtration on a Sephadex 75 10/300 GL column. SAL was found to be a monomer of 39916.3 Da, as determined by tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance liquid chromatography (HPLC). The N-terminal 10-amino acid sequence of SAL, KPWALSFSFG, resembles those of other legume lectins. SAL exhibits hemagglutinating activity against rabbit erythrocytes at 11.9 μg/ml. Its hemagglutinating activity is stable in the pH range 7-11 and in the temperature range 30-90°C, and is stimulated by Mn(2+). The hemagglutinating activity of SAL is most potently inhibited by 50-mM d-galactose. SAL suppresses mycelial growth in Penicillium digitatum and Alternaria alternata; the IC(50) of the antifungal activity toward P. digitatum and A. alternata were found to be 3.125 and 3.338 μM, respectively. SAL suppresses the proliferation of human cervical cancer cells (HeLa) at an IC(50) of 6.25 μM (P< 0.05). But it has no inhibiting effect on bacteria. This is the first report of a lectin from seeds of S. alopecuroides. PMID:22634632

  15. Lectin-Glycan Interaction Network-Based Identification of Host Receptors of Microbial Pathogenic Adhesins

    Science.gov (United States)

    Ielasi, Francesco S.; Alioscha-Perez, Mitchel; Donohue, Dagmara; Claes, Sandra; Sahli, Hichem; Schols, Dominique

    2016-01-01

    ABSTRACT The first step in the infection of humans by microbial pathogens is their adherence to host tissue cells, which is frequently based on the binding of carbohydrate-binding proteins (lectin-like adhesins) to human cell receptors that expose glycans. In only a few cases have the human receptors of pathogenic adhesins been described. A novel strategy—based on the construction of a lectin-glycan interaction (LGI) network—to identify the potential human binding receptors for pathogenic adhesins with lectin activity was developed. The new approach is based on linking glycan array screening results of these adhesins to a human glycoprotein database via the construction of an LGI network. This strategy was used to detect human receptors for virulent Escherichia coli (FimH adhesin), and the fungal pathogens Candida albicans (Als1p and Als3p adhesins) and C. glabrata (Epa1, Epa6, and Epa7 adhesins), which cause candidiasis. This LGI network strategy allows the profiling of potential adhesin binding receptors in the host with prioritization, based on experimental binding data, of the most relevant interactions. New potential targets for the selected adhesins were predicted and experimentally confirmed. This methodology was also used to predict lectin interactions with envelope glycoproteins of human-pathogenic viruses. It was shown that this strategy was successful in revealing that the FimH adhesin has anti-HIV activity. PMID:27406561

  16. Impact of Mannose-Binding Lectin Deficiency on Radiocontrast-Induced Renal Dysfunction

    Directory of Open Access Journals (Sweden)

    Michael Osthoff

    2013-01-01

    Full Text Available Contrast-induced nephropathy (CIN is the third leading cause of acute renal failure in hospitalized patients. Endothelial dysfunction, renal medullary ischemia, and tubular toxicity are regarded as the most important factors in the pathogenesis of CIN. Mannose-binding lectin (MBL, a pattern recognition protein of the lectin pathway of complement, has been found to aggravate and mediate tissue damage during experimental renal ischemia/reperfusion (I/R injury which was alleviated by inhibition with C1 inhibitor, a potent MBL, and lectin pathway inhibitor. In this paper, we highlight the potential role of MBL in the pathogenesis of human CIN. In experimental I/R models, MBL was previously found to induce tubular cell death independent of the complement system. In addition, after binding to vascular endothelial cells, MBL and its associated serine proteases were able to trigger a proinflammatory reaction and contribute to endothelial dysfunction. In humans, urinary MBL was increased after administration of contrast media and in individuals with CIN. Moreover, individuals with normal/high MBL levels were at increased risk to develop radiocontrast-induced renal dysfunction. Hence, MBL and the lectin pathway seem to be a promising target given that a licensed, powerful, human recombinant inhibitor exits to be added to the scarce armamentarium currently available for prophylaxis of CIN.

  17. Lectin-binding characteristics of a Lyme borreliosis spirochete Borrelia burgdorferi sensu stricto

    Czech Academy of Sciences Publication Activity Database

    Vancová, M.; Nebesářová, J.; Grubhoffer, Libor

    2005-01-01

    Roč. 50, č. 3 (2005), s. 229-238. ISSN 0015-5632 R&D Projects: GA ČR GA206/03/1323; GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia burgdorferi * electron microscopy * lectin binding Subject RIV: EE - Microbiology, Virology Impact factor: 0.918, year: 2005

  18. Ficolin-3-mediated lectin complement pathway activation in patients with subarachnoid hemorrhage

    DEFF Research Database (Denmark)

    Zanier, Elisa R; Zangari, Rosalia; Munthe-Fog, Lea;

    2014-01-01

    OBJECTIVES: To assess the involvement of ficolin-3, the main initiator of the lectin complement pathway (LCP), in subarachnoid hemorrhage (SAH) pathology and outcome. METHODS: In this preliminary exploratory study, plasma concentration of ficolin-3 and of ficolin-3-mediated functional LCP activit...

  19. Carbohydrate Detection and Lectin Isolation from Tegumental Tissue of Fasciola hepatica

    Directory of Open Access Journals (Sweden)

    MB Molaei Rad

    2010-02-01

    Full Text Available "nBackground: Fascioliasis is a chronic hepatic disease and may be resulted from mechani­cal/molecular parasite adhesion to host liver tissue. The aim of this study was to detect surface car­bohydrate and lectin, carbohydrate-binding protein isolation that might be responsible of this molecular binding."nMethods: The present experimental work was conducted in the Department of Medical Parasitol­ogy and Mycology, School of Public Health, Tehran University of Medical Sciences, Te­hran, Iran.  Fasciola hepatica parasites were collected from abattoir (Saman, Tehran, Iran and surface mannose-carbohydrate was detected by fluorescein isothiocyanate (FITC conju­gated lectin (Lentil. Lectin of tegumental tissue from F. hepatica was isolated by affinity chroma­tography and detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE."nResults: Mannose carbohydrate was observed on the surface of tegumental tissue from para­site under fluorescence microscope. Carbohydrate-binding protein or lectin with MW of 50 kDa also was isolated from homogenized tegument of helminth."nConclusion: These results are important for understanding of molecular pathogenesis of F. hepat­ica at the chronic phase of fascioliasis

  20. Two lectin-like receptors for alpha 1-acid glycoprotein in mouse testis

    DEFF Research Database (Denmark)

    Andersen, U O; Kirkeby, S; Bøg-Hansen, T C

    Sertoli cells and, at the last stages in the spermatogenic cycle, a very strong reaction in the late elongated spermatids and the apical extensions of Sertoli cells. The interactions are lectin-like as confirmed by inhibition with simple sugars. In addition, the bindings were inhibited by steroid hormones...

  1. Lectin histochemistry of Kudoa septempunctata genotype ST3-infected muscle of olive flounder (Paralichthys olivaceus).

    Science.gov (United States)

    Kang, Jaeyoun; Park, Changnam; Jang, Yeounghwan; Ahn, Meejung; Shin, Taekyun

    2016-01-01

    The localization of carbohydrate terminals in Kudoa septempunctata ST3-infected muscle of olive flounder (Paralichthys olivaceus) was investigated using lectin histochemistry to determine the types of carbohydrate sugar residues expressed in Kudoa spores. Twenty-one lectins were examined, i.e., N-acetylglucosamine (s-WGA, WGA, DSL-II, DSL, LEL, STL), mannose (Con A, LCA, PSA), galactose/N-acetylgalactosamine (RCA12, BSL-I, VVA, DBA, SBA, SJA, Jacalin, PNA, ECL), complex type N-glycans (PHA-E and PHA-L), and fucose (UEA-I). Spores encased by a plasmodial membrane were labeled for the majority of these lectins, with the exception of LCA, PSA, PNA, and PHA-L. Four lectins (RCA 120, BSL-I, DBA, and SJA) belonging to the galactose/N-acetylgalactosamine group, only labeled spores, but not the plasmodial membrane. This is the first confirmation that various sugar residues are present in spores and plasmodial membranes of K. septempunctata ST3. PMID:27169676

  2. Lectin histochemistry of Kudoa septempunctata genotype ST3-infected muscle of olive flounder (Paralichthys olivaceus

    Directory of Open Access Journals (Sweden)

    Kang Jaeyoun

    2016-01-01

    Full Text Available The localization of carbohydrate terminals in Kudoa septempunctata ST3-infected muscle of olive flounder (Paralichthys olivaceus was investigated using lectin histochemistry to determine the types of carbohydrate sugar residues expressed in Kudoa spores. Twenty-one lectins were examined, i.e., N-acetylglucosamine (s-WGA, WGA, DSL-II, DSL, LEL, STL, mannose (Con A, LCA, PSA, galactose/N-acetylgalactosamine (RCA12, BSL-I, VVA, DBA, SBA, SJA, Jacalin, PNA, ECL, complex type N-glycans (PHA-E and PHA-L, and fucose (UEA-I. Spores encased by a plasmodial membrane were labeled for the majority of these lectins, with the exception of LCA, PSA, PNA, and PHA-L. Four lectins (RCA 120, BSL-I, DBA, and SJA belonging to the galactose/N-acetylgalactosamine group, only labeled spores, but not the plasmodial membrane. This is the first confirmation that various sugar residues are present in spores and plasmodial membranes of K. septempunctata ST3.

  3. Crystallization and preliminary X-ray characterization of a lectin from Cicer arietinum (chickpea)

    International Nuclear Information System (INIS)

    The crystallization and characterization of a lectin isolated and purified from C. arietinum and possessing complex sugar specificity is reported. The lectin isolated from mature seeds of Cicer arietinum (CAL) agglutinates pronase-treated rabbit and human erythrocytes and its haemagglutination activity is inhibited by fetuin and desialated fetuin but not by simple monosaccharides or oligosaccharides. The purified lectin is a dimer of molecular weight 43 000 Da composed of two identical subunits (MW 21 500), as confirmed by SDS–PAGE. The lectin has been crystallized using the hanging-drop vapour-diffusion method at 295 K over a well solution containing 0.2 M sodium acetate, 0.1 M sodium phosphate buffer pH 6.5 and 14%(w/v) polyethylene glycol 8000. The triangular prism-shaped crystals belong to space group R3 and have unit-cell parameters a = b = 81.2, c = 69.4 Å. The diffraction data are 93.8% complete to 2.3 Å Bragg spacing with an Rmerge of 0.103

  4. Crystallization and preliminary X-ray characterization of a lectin from Cicer arietinum (chickpea)

    Energy Technology Data Exchange (ETDEWEB)

    Katre, Uma V.; Gaikwad, S. M. [Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008 (India); Bhagyawant, S. S.; Deshpande, U. D. [School of Life Sciences, S. R. T. M. University, Nanded 431606 (India); Khan, M. I.; Suresh, C. G., E-mail: suresh@ems.ncl.res.in [Division of Biochemical Sciences, National Chemical Laboratory, Pune 411008 (India)

    2005-01-01

    The crystallization and characterization of a lectin isolated and purified from C. arietinum and possessing complex sugar specificity is reported. The lectin isolated from mature seeds of Cicer arietinum (CAL) agglutinates pronase-treated rabbit and human erythrocytes and its haemagglutination activity is inhibited by fetuin and desialated fetuin but not by simple monosaccharides or oligosaccharides. The purified lectin is a dimer of molecular weight 43 000 Da composed of two identical subunits (MW 21 500), as confirmed by SDS–PAGE. The lectin has been crystallized using the hanging-drop vapour-diffusion method at 295 K over a well solution containing 0.2 M sodium acetate, 0.1 M sodium phosphate buffer pH 6.5 and 14%(w/v) polyethylene glycol 8000. The triangular prism-shaped crystals belong to space group R3 and have unit-cell parameters a = b = 81.2, c = 69.4 Å. The diffraction data are 93.8% complete to 2.3 Å Bragg spacing with an R{sub merge} of 0.103.

  5. Mannose-binding lectin and infection risk in newborns: a systematic review

    NARCIS (Netherlands)

    J. Israëls; F.N.J. Frakking; L.C.M. Kremer; M. Offringa; T.W. Kuijpers; M.D. van de Wetering

    2010-01-01

    The authors systematically reviewed the literature on mannose-binding lectin (MBL) and infections in newborns to determine whether infection risk is increased in MBL-deficient newborns. All original reports on MBL and infections in newborns were retrieved from Embase, Medline and CENTRAL from 1966 t

  6. A survey of domestic species of Basidiomycetes fungi for the presence of lectins inn their carpophores

    Directory of Open Access Journals (Sweden)

    Grażyna Końska

    2014-02-01

    Full Text Available Preliminary investigations were conducted to determine the presence of active lectins in carpophores of fungi from the class Basidiomycetes, collected from natural localities in southern and south-eastern Poland. The degree of agglutination activity (expressed as the titre of agglutination of aqueous extracts was determined at room temperature (18-20°C and at +4°C in respect to human and animal erythrocytes suspended in physiological saline, part of which were additionally treated with proteolytic enzymes. From among the 104 tested species, extracts from 41 of them showed agglutination activity, among which 18 were high. In six cases, specific activity against human ABH group antigens was found. Extracts from 5 species agglutinated only animal erythrocytes, with pigeon erythrocytes being exceptionally sensitive to the lectins. Extracts from two species had distinctly higher agglutination activity at 4°C, which suggests that lectins of the "cold" agglutinin type are present in these species. Analysis of extracts from caps and stems showed that caps had a higher lectin content.

  7. Magnetic ovalbumin and egg white aggregates as affinity adsorbents for lectins separation

    Czech Academy of Sciences Publication Activity Database

    Maděrová, Zdeňka; Šafaříková, Miroslava; Šafařík, Ivo

    2008-01-01

    Roč. 40, - (2008), s. 542-545. ISSN 1369-703X R&D Projects: GA MŠk OC 157; GA MPO 2A-1TP1/094 Institutional research plan: CEZ:AV0Z60870520 Keywords : egg white * lectins * magnetic protein aggregates Subject RIV: EI - Biotechnology ; Bionics Impact factor: 1.889, year: 2008

  8. "Role of Lectins in Interaction Between Parasites and the Important Insect Vectors"

    Directory of Open Access Journals (Sweden)

    HR Basseri

    2002-08-01

    Full Text Available There is growing evidence that lectin-carbohydrate interactions can mediate the infection of parasites to their insect vector. Many insect species are host or vectors of protozoan or metazoan parasites that cause socially and economically important disease such as malaria and leishmaniasis. However, relatively little work has been undertaken concerning the interaction of insect immunity against parasite invasion with respect to lectins activities. Both immune defences (cellular and noncellular of insect haemolymph react in order to combat the diverse array of natural pathogens and other microorganisms. The most of immune substances are innate, naturally-occurring and nonspecific molecules present in haemolymph. When the physical defences of the insect gut or integument are breached by an invading organism an innate response begins, characterized by immune system's agents such as coagulation, melanization, phagocytosis, encapsulation and nodule formation. Nevertheless, in many cell types such as insect haemocytes, carbohydrates are known to be crucially involved in cell-cell interactions and many studies have addressed the role of carbohydrates and carbohydratebinding molecules in the adhesion of parasites to their host. As mentioned above, one candidate for attachment and invasion may be lectins or lectinlike molecules that are known to mediate cell-to-cell interaction. In order to the basic understanding of pathogens transmission by vectors, in this article, the interaction between parasites and insect vectors has been reviewed with respect to role of lectins molecules.

  9. Lectin histochemistry of Kudoa septempunctata genotype ST3-infected muscle of olive flounder (Paralichthys olivaceus)

    Science.gov (United States)

    Kang, Jaeyoun; Park, Changnam; Jang, Yeounghwan; Ahn, Meejung; Shin, Taekyun

    2016-01-01

    The localization of carbohydrate terminals in Kudoa septempunctata ST3-infected muscle of olive flounder (Paralichthys olivaceus) was investigated using lectin histochemistry to determine the types of carbohydrate sugar residues expressed in Kudoa spores. Twenty-one lectins were examined, i.e., N-acetylglucosamine (s-WGA, WGA, DSL-II, DSL, LEL, STL), mannose (Con A, LCA, PSA), galactose/N-acetylgalactosamine (RCA12, BSL-I, VVA, DBA, SBA, SJA, Jacalin, PNA, ECL), complex type N-glycans (PHA-E and PHA-L), and fucose (UEA-I). Spores encased by a plasmodial membrane were labeled for the majority of these lectins, with the exception of LCA, PSA, PNA, and PHA-L. Four lectins (RCA 120, BSL-I, DBA, and SJA) belonging to the galactose/N-acetylgalactosamine group, only labeled spores, but not the plasmodial membrane. This is the first confirmation that various sugar residues are present in spores and plasmodial membranes of K. septempunctata ST3. PMID:27169676

  10. Analysis of trypsin inhibitors and lectins in white kidney beans (Phaseolus vulgaris, var. Processor) in a combined method.

    Science.gov (United States)

    Roozen, J P; de Groot, J

    1991-01-01

    Buffered saline extraction, affinity chromatography, and Folin-BSA protein assay were used consecutively to provide a combined method for analysis of trypsin inhibitors and lectins in white kidney beans (Phaseolus vulgaris, var. Processor). The method was tested by following the decrease of both antinutritional factors by germination of the beans for 7 days at 20 degrees C. Repeatability coefficients of variation were 2-7.4% for the trypsin inhibitors and 2.2-10% for the lectins. After 7 days of germination, trypsin inhibitors and lectins were reduced by 72 and 92%, respectively. PMID:1757418

  11. Technical Report on the Development of Novel Technology for Reducing the Toxicity of Mistletoe Lectin by using Radiation Fusion Technology

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Ju Woon; Kim, Jae Hun; Choi, Jong Il; Song, Beom Seok; Yoon, Yo Han; Jung, Pil Mun; Sung, Nak Yun

    2009-10-15

    The aim of this study was conducted to investigate the effect of irradiation on detoxification, structural change, and physiological change of Mistletoe lectin. Optimal irradiation dose was determined from the result of having maximum detoxification and remaining the immunological activity Irradiation technology could be effective method for detoxification of Mistletoe lectin containing the immunological activity. The results indicate the feasibility of novel technology for reduction of the toxicity of Mistletoe lectin by using radiation technology. Practical state though clinical test is needed to extend biomedicine field using radiation technology and improve of public health by the control of the disease that gradually increase every year.

  12. Technical Report on the Development of Novel Technology for Reducing the Toxicity of Mistletoe Lectin by using Radiation Fusion Technology

    International Nuclear Information System (INIS)

    The aim of this study was conducted to investigate the effect of irradiation on detoxification, structural change, and physiological change of Mistletoe lectin. Optimal irradiation dose was determined from the result of having maximum detoxification and remaining the immunological activity Irradiation technology could be effective method for detoxification of Mistletoe lectin containing the immunological activity. The results indicate the feasibility of novel technology for reduction of the toxicity of Mistletoe lectin by using radiation technology. Practical state though clinical test is needed to extend biomedicine field using radiation technology and improve of public health by the control of the disease that gradually increase every year

  13. Activation of the lectin pathway by natural IgM in a model of ischemia/reperfusion injury

    DEFF Research Database (Denmark)

    Zhang, M.; Takahashi, K.; Alicot, E.M.;

    2006-01-01

    Reperfusion of ischemic tissues elicits an acute inflammatory response involving serum complement, which is activated by circulating natural IgM specific to self-Ags exposed by ischemia. Recent reports demonstrating a role for the lectin pathway raise a question regarding the initial events in...... complement activation. To dissect the individual roles of natural IgM and lectin in activation of complement, mice bearing genetic deficiency in early complement, IgM, or mannan-binding lectin were characterized in a mesenteric model of ischemia reperfusion injury. The results reveal that IgM binds initially...

  14. Characterization of a lectin from the craysfish Cherax quadricarinatus hemolymph and its effect on hemocytes.

    Science.gov (United States)

    Sánchez-Salgado, J L; Pereyra, M A; Vivanco-Rojas, O; Sierra-Castillo, C; Alpuche-Osorno, J J; Zenteno, E; Agundis, C

    2014-08-01

    Lectins participate in the immune mechanisms of crustaceans. They have been considered as humoral receptors for pathogen-associated molecular patterns; however, some reports suggest that lectins could regulate crustacean cellular functions. In the present study, we purified and characterized a serum lectin (CqL) from the hemolymph of Cherax quadricarinatus by affinity chromatography and determined its participation in the regulation of hemocytes' oxidative burst. CqL is a 290-kDa lectin in native form, constituted by 108, 80, and 29-kDa subunits. It is mainly composed of glycine, alanine, and a minor proportion of methionine and histidine. It showed no carbohydrates in its structure. CqL is composed of several isoforms, as determined by 2D-electrophoresis, and shows no homology with any crustacean protein as determined by Lc/Ms mass spectrometry. CqL agglutinated mainly rat and rabbit erythrocytes and showed a broad specificity for monosaccharides such as galactose, glucose, and sialic acid, as well as for glycoproteins, such as porcine stomach and bovine submaxillary mucin and fetuin. It is a Mn(2+)-dependent lectin. CqL recognized 8% of crayfish granular hemocytes and increased 4.2-fold the production of hemocytes' superoxide anion in vitro assays when compared with non-treated hemocytes. This effect showed the same specificity for carbohydrates as hemagglutination; moreover, superoxide dismutase and diphenyleneiodonium chloride were effective inhibitors of CqL oxidative-activation. The CqL homoreceptor is a 120-kDa glycoprotein identified in the hemocytes lysate. Our results suggest that CqL participates actively in the regulation of the generation of superoxide anions in hemocytes using NADPH-dependent mechanisms. PMID:24929243

  15. Molecular Characterization and Global Expression Analysis of Lectin Receptor Kinases in Bread Wheat (Triticum aestivum)

    Science.gov (United States)

    Pandey, Ajay K.; Singh, Kashmir; Upadhyay, Santosh Kumar

    2016-01-01

    Lectin receptor kinases (LRKs) play a critical role in plants during development and stress conditions, but a comprehensive analysis at genome level is still not carried out in Triticum aestivum. Herein, we performed the genome wide identification, characterization and expression analysis of these genes in T. aestivum (TaLRK). In-total 263 TaLRK genes were identified, which were further classified into three groups based on the nature of lectin domain. We identified, two TaLRKs consisted of calcium-dependent lectin (C-LRK), while 84 legume-lectin (L-LRK) and 177 bulb-lectin (B-LRK) domains. The L-LRK and B-LRK genes were distributed throughout the genome of T. aestivum. Most of the TaLRKs were clustered as homologs, which were distributed either in proximity on same chromosome or on homoeologous chromosomes of A, B and D sub-genomes. A total of 9 and 58 duplication events were also predicted in L-LRK and B-LRK, respectively. Phylogenetic analysis indicated conserved evolutionary relationship of homologous and orthologous genes from multiple plant species. Gene ontology analysis indicated TaLRKs role in binding, signaling and receptor activities. Most of the TaLRKs consisted of a trans-membrane domain and predicted to be localized in the plasma-membrane. A diverse expression pattern of TaLRK genes was found in various developmental stages and stress conditions. Some TaLRKs were found to be highly affected during a particular stress, which indicated a specialized role of each LRK gene in a specific stress condition. These results described various characteristic feature and expression pattern of TaLRK genes, which will pave the way for functional characterization in wheat. PMID:27111449

  16. Three-dimensional structure of lectin from Dioclea violacea and comparative vasorelaxant effects with Dioclea rostrata

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, B.A.M.; Bezerra, M.J.B.; Bezerra, G.A.; Alencar, K.L.L.; Nascimento, K.S.; Naganao, C.S.; Sampaio, A.H.; Cavada, B.S. [Universidade Federal do Ceara (UFC), Fortaleza, CE (Brazil); Delatorre, P. [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil); Rodrigues, N.V.; Pires, A.F.; Assreuy, A.M.S. [Universidade Estadual do Ceara (UECE), Fortaleza, CE (Brazil); Marins, J.L. [Universidade Federal de Pelotas (UFPel), Pelotas, RS (Brazil)

    2012-07-01

    Full text: Lectins are a structural heterogeneous group of proteins possessing at least one non-catalytic domain that binds reversibly to a specific mono or oligosaccharide. Diocleinae lectins exhibit glucose/mannose monosaccharide binding specificity and studies of their chemical and physicochemical properties revealed a high degree of identity in their amino acid sequences and three dimensional structures. This study investigated structural/functional relationships between lectins obtained from Dioclea violacea (DVL) and Dioclea rostrata (DRL). The purified lectin (DVL) was solubilized in 20 mM Tris-HCl pH 7.6 with 5 mM CaCl{sub 2} and MnCl{sub 2} buffer and incubated during one hour before the crystallization experiments with the ligand X-Man (5-bromo-4-chloro-3-indolyl-{alpha}-D-mannose) at 3 mM. Crystals of DVL grew in condition 33 of Crystal Screen I (4M Sodium formate) and belong to the orthorhombic space group I222. The structure of DVL at 2.6 resolution was obtained by molecular replacement using the coordinates of DRL (PDB code 2ZBJ), after the last refinement the structure presented R factor of 0.23 and R free of 0.27. The crystal structures reveal differences between them and could be related to relaxant activity. The conformation of residues HIS51, HIS131 and GLU205 and others positioned at CRD lead to different lectin binding activities. In fact, the pocket in DVL is small and deep and promotes weak interaction with carbohydrates, while DRL pocket is large and shallow, allowing strong interaction between CRD and sugars. This can explain why DVL and DRL elicited different degrees of aorta relaxation showing maximal effects of 43 % and 96 %, respectively. (author)

  17. Distribution of lectin-bindings in the testis of the lesser mouse deer, Tragulus javanicus.

    Science.gov (United States)

    Agungpriyono, S; Kurohmaru, M; Kimura, J; Wahid, A H; Sasaki, M; Kitamura, N; Yamada, J; Fukuta, K; Zuki, A B

    2009-06-01

    The distribution of lectin bindings in the testis of the smallest ruminant, lesser mouse deer (Tragulus javanicus), was studied using 12 biotinylated lectins specific for d-galactose (peanut agglutinin PNA, Ricinus communis agglutinin RCA I), N-acetyl-d-galactosamine (Dolichos biflorus agglutinin DBA, Vicia villosa agglutinin VVA, Soybean agglutinin SBA), N-acetyl-D-glucosamine and sialic acid (wheat germ agglutinin WGA, s-WGA), D-mannose and d-glucose (Lens culinaris agglutinin LCA, Pisum sativum agglutinin PSA, Concanavalin A Con A), L-fucose (Ulex europaeus agglutinin UEA I), and oligosaccharide (Phaseolus vulgaris agglutinin PHA-E) sugar residues. In Golgi-, cap-, and acrosome-phase spermatids, lectin-bindings were found in the acrosome (PNA, RCA I, VVA, SBA, WGA and s-WGA), and in the cytoplasm (PNA, RCA I, VVA, SBA, WGA, LCA, PSA, Con A and PHA-E). s-WGA binding was confined to the spermatid acrosome, but other lectins were also observed in spermatocytes. In spermatogonia, VVA, WGA, Con A, and PHA-E bindings were observed. Sertoli cells were intensely stained with DBA and Con A, and weakly with PHA-E. In interstitial Leydig cells, RCA I, DBA, VVA, Con A, PSA, LCA, WGA and PHA-E were positive. UEA I was negative in all cell types including spermatogenic cells. Unusual distribution of lectin-bindings noted in the testis of lesser mouse deer included the limited distribution of s-WGA only in the spermatid acrosome, the distribution of DBA in Sertoli cells, Leydig cells and lamina propria, and the absence of UEA I in all type cells. The present results were discussed in comparison with those of other animals and their possible functional implications. PMID:19245668

  18. First isolation and characterization of a novel lectin with potent antitumor activity from a Russula mushroom.

    Science.gov (United States)

    Zhang, G; Sun, J; Wang, H; Ng, T B

    2010-08-01

    To date only a ribonuclease and a protein with anti-HIV-1 reverse transcriptase activity have been isolated from mushrooms of the genus Russula. In this study a novel lectin, with a molecular weight of 32 kDa, and a unique N-terminal sequence different from other lectins, was isolated from the mushroom Russula lepida. It represents the first lectin isolated from Russula mushrooms. The purification scheme involved (NH4)2SO4 precipitation, ion exchange chromatography on diethylaminoethyl DEAE-cellulose and SP-Sepharose, and fast protein liquid chromatography-gel filtration on Superdex 75. The hemagglutinating activity of the lectin (RLL) was inhibited by inulin and O-nitrophenyl-beta-D-galacto-pyranoside. The lectin was stable at temperatures up to 70 degrees C (half of the activity was preserved at 80 degrees C), and in the presence of NaOH or HCl solutions up to a concentration of 12.5 mM. Its hemagglutinating activity was reduced in the presence of Mn2+, Co2+, and Hg2+ ions, and enhanced by Cu2+ ions. It exhibited antiproliferative activity toward hepatoma Hep G2 cells and human breast cancer MCF-7 cells with an IC(50) of 1.6 microM and 0.9 microM, respectively. Daily intraperitoneal injections of RLL (5.0 mg/kg body weight/day for 20 days) brought about 67.6% reduction in the weight of S-180 tumor. RLL was devoid of antifungal, ribonuclease, and HIV-1 reverse transcriptase inhibitory activities. PMID:20378319

  19. Carbohydrate/glycan-binding specificity of legume lectins in respect to their proposed biological functions

    Directory of Open Access Journals (Sweden)

    Márcio Viana Ramos

    2000-01-01

    Full Text Available The lectins, proteins which specifically recognize carbohydrate moieties, have been extensively studied in many biochemical and structural aspects in order to establish the molecular basis of this non-catalytic event. On the other hand, their clinical and agricultural potentials have been growing fast. Although lectins, mainly those from legume plants, had been investigated for biological properties, studies about the physiological functions of lectins are scarce in literature. Therefore, despite the accumulated data on lectins (as proteins, the role played by these signalizing molecules is poorly discussed. In the light of our accumulated results on legume lectins, specially those obtained from plants belonging to the Diocleinae sub-tribe and available data in literature, we discuss here the main hypothesis of their functions according to their carbohydrate/glycan-binding specificity.As lectinas, proteinas que especificamente reconhecem estruturas que contém carboidratos, têm sido extensivamente estudadas em muitos aspectos bioquímicos e estruturais, objetivando estabelecer as bases moleculares deste evento não-catalítico. Por outro lado, os potenciais clínicos e agriculturais destas proteínas têm crescido rapidamente. Embora as lectinas, principalmente aquelas de legumes tenham sido bastante investigadas em suas propriedades biológicas, estudos sobre as funcões fisiológicas de lectinas são escassos na literatura. Além disto, a despeito da quantidade de dados acumulados sobre lectinas (como proteínas, o papel desempenhado por estas moléculas de sinalização é pobremente discutido. Valendo-se de nossos estudos sobre lectinas de leguminosas, principalmente da sub-tribo Diocleinae, e outros dados presentes na literatura, discutimos aqui, as principais hipóteses de suas funções com base na especificidade por carboidratos e glicanos complexos.

  20. Three-dimensional structure of lectin from Dioclea violacea and comparative vasorelaxant effects with Dioclea rostrata

    International Nuclear Information System (INIS)

    Full text: Lectins are a structural heterogeneous group of proteins possessing at least one non-catalytic domain that binds reversibly to a specific mono or oligosaccharide. Diocleinae lectins exhibit glucose/mannose monosaccharide binding specificity and studies of their chemical and physicochemical properties revealed a high degree of identity in their amino acid sequences and three dimensional structures. This study investigated structural/functional relationships between lectins obtained from Dioclea violacea (DVL) and Dioclea rostrata (DRL). The purified lectin (DVL) was solubilized in 20 mM Tris-HCl pH 7.6 with 5 mM CaCl2 and MnCl2 buffer and incubated during one hour before the crystallization experiments with the ligand X-Man (5-bromo-4-chloro-3-indolyl-α-D-mannose) at 3 mM. Crystals of DVL grew in condition 33 of Crystal Screen I (4M Sodium formate) and belong to the orthorhombic space group I222. The structure of DVL at 2.6 resolution was obtained by molecular replacement using the coordinates of DRL (PDB code 2ZBJ), after the last refinement the structure presented R factor of 0.23 and R free of 0.27. The crystal structures reveal differences between them and could be related to relaxant activity. The conformation of residues HIS51, HIS131 and GLU205 and others positioned at CRD lead to different lectin binding activities. In fact, the pocket in DVL is small and deep and promotes weak interaction with carbohydrates, while DRL pocket is large and shallow, allowing strong interaction between CRD and sugars. This can explain why DVL and DRL elicited different degrees of aorta relaxation showing maximal effects of 43 % and 96 %, respectively. (author)

  1. [L-fucose-specific lectin from pike perch (Lucioperca lucioperca L.) roe. Purification and studies of carbohydrate specificity].

    Science.gov (United States)

    Antoniuk, V O

    2004-01-01

    L-fucose-specific lectin was purified from pike perch (Lucioperca lucioperca L.) roe by affinity chromatography on ovariomucine H-sepharose from a human ovary cyst. Three bands were detected after disk-electrophoresis in PAAG in alkaline (pH 8.9) and five bands--in acidic system (pH 4.3). According to electrophoresis data in 15% SDS-PAGE the lectin contains two components with molecular weight 13-14 kDa. Molecular weight of the lectin is 50 kDa according to gel-chromatography on tojopearl HW-55. The immunochemical properties of the lectins from perch (Persa fluviatilis L.) roe and pike perch (Lucioperca lucioperca L.) roe are similar. PMID:15915715

  2. Cellular carbohydrate components in human, rabbit and rat lacrimal gland. Studies using fluorescein and peroxidase labelled lectins.

    Science.gov (United States)

    Ahmed, A; Grierson, I

    1989-01-01

    Orbital lacrimal glands from adult male and female rabbits, rats and humans were examined for the presence of intracellular receptors of four lectins: concanavalin-A agglutinin, lutus tetragonolobus agglutinin, ricinus comunis-60 agglutinin and wheat-germ agglutinin using fluorescein-conjugated lectin and peroxidase labelling methods for fluorescence and electron microscopy, respectively. Lectins were used as specific probes to detect carbohydrate moiety of the lacrimal gland. The pattern of labelling with the lectins suggests that N-acetyl-glucosamine, N-acetyl-D-galactosamine, D-galactose, D-mannose, sialic acid and L-fucose are contained in the lacrimal gland of the three species. The significance of these findings is discussed. PMID:2920911

  3. Microarray-based identification of lectins for the purification of human urinary extracellular vesicles directly from urine samples.

    Science.gov (United States)

    Echevarria, Juan; Royo, Felix; Pazos, Raquel; Salazar, Lorena; Falcon-Perez, Juan Manuel; Reichardt, Niels-Christian

    2014-07-21

    As cellular-derived vesicles largely maintain the biomolecule composition of their original tissue, exosomes, which are found in nearly all body fluids, have enormous potential as clinical disease markers. A major bottleneck in the development of exosome-based diagnostic assays is the challenging purification of these vesicles; this requires time-consuming and instrument-based procedures. We employed lectin arrays to identify potential lectins as probes for affinity-based isolation of exosomes from the urinary matrix. We found three lectins that showed specific interactions to vesicles and no (or only residual) interaction with matrix proteins. Based on these findings a bead-based method for lectin-based isolation of exosomes from urine was developed as a sample preparation step for exosome-based biomarker research. PMID:25044519

  4. [Comparative assessment of inductive effects of Azospirillum lectins with different antigenic properties on the signal systems of wheat seedling roots].

    Science.gov (United States)

    Alen'kina, S A; Petrova, L P; Sokolova, M K; Chernyshova, M P; Trutneva, K A; Bogatyrev, V A; Nikitina, V E

    2014-01-01

    The lectins of associative nitrogen-fixing bacteria Azospirillum brasilense Sp7 and its mutant A. brasilense Sp7.2.3 were shown to have different effects on the components of the wheat seedling root signal system, namely to regulate the levels of cAMP, nitric oxide, diacylglycerol, and salicylic acid, as well as to induce the activities of superoxide dismutase and lipoxygenase. Our results make it possible to consider azospirilla lectins as inducers of the signal systems in wheat seedling roots, since they cause development of several flows of primary signals. These data are of general biological importance, since lectins are present in all living organisms and most ot the functions of lectins remain insufficiently understood. PMID:25844444

  5. Hayabusa's follow-on mission for surface and sub-surface sample return from a C-type NEO

    Science.gov (United States)

    Yano, Hajime; Yoshikawa, M.; Yano, H.; Tsuda, Y.; Nakazawa, S.; Mimamino, H.; Terui, F.; Saiki, T.; Nishiyama, K.; Kubota, T.; Okada, T.; Morimoto, M. Y.; Ogawa, N.; Okamoto, C.; Takagi, Y.; Tachibana, S.; Nakamura, R.; Hirata, N.; Demura, H.

    n JAXA's Long-term Vision 2005-2025, acquiring the capability of deep space round trip be-yond the Earth-Moon system is one of key elements for the future space exploration and that has been Hayabusa's primary engineering goal. According to the solar system exploration sci-ence roadmap set by ISAS and JSPEC in 2007, a programmatic approach to small body sample returns from S-type, C-type and then P/D-type asteroids as well as dormant comets, i.e., 'the further, the smaller, the more primitive strategy', is recommended for strengthening Japan's unique position in the field of space exploration. In a more recent international context, NEOs and Martian satellites have been identified as critical targets for the future human space explo-ration en route to Mars; thus their robotic precursor missions with the round trip capability have become more important than ever. Thus, Hayabusa's immediate follow-on mission, nicknamed so far as 'Hayabusa-2', is to aim establishing round trip exploration capability with both technical and operational heritage and lessons leaerned from the original Haybusa mission. It will also conduct in-situ observation and surface and sub-surface sample returns of a C-type NEO after Hayabusa's investigation and sampling attempt at Itokawa, a sub-km, S-type NEO. Important to be reminded is that C-type asteroid exploration is not just matching with carbona-ceous chondrites and interplanetary dust but also enhancing chances to discover new extrater-restrial materials unknown to us today that may become clues to decode interactions among organic, inorganic compounds and "water" kept in various forms inside the object. These three groups of asteroidal materials are basic constituents of the planet Earth, its ocean and its life. Also physical probing inside solid planetary bodies has been recognized as an effective tool to open new scientific insights. By excavating sub-surface materials with artificial physical in-teractions such as an impactor

  6. Targeting the Cryptococcus neoformans var. grubii Cell Wall Using Lectins: Study of the Carbohydrate-Binding Domain

    OpenAIRE

    Pamella de Brito Ximenes; Eduardo Isidoro Carneiro Beltrão; Danielle Patrícia Cerqueira Macêdo; Maria Daniela Silva Buonafina; Reginaldo Gonçalves de Lima-Neto; Rejane Pereira Neves

    2015-01-01

    Cryptococcus neoformans var. grubii is considered to be the major cause of cryptococcosis in immunosuppressed patients. Understanding cell wall glycoproteins using lectins is of medical interest and can contribute to specific therapy. The aim of this study was to evaluate the carbohydrates on the cell wall of Cryptococcus neoformans var. grubii clinical isolates, using a fluorescein isothiocyanate-lectin binding protocol. Thirty yeast strains stocked in the culture collection were cultivated ...

  7. Effect of bacterial lectin on acceleration of fat cell lipolysis at in vitro diode laser treatment using encapsulated ICG

    Science.gov (United States)

    Yanina, Irina Yu.; Kochubey, Vyacheslav I.; Tuchin, Valery V.; Portnov, Sergey A.; Svenskaya, Yuliya I.; Gorin, Dmitry A.; Ponomareva, Elena G.; Nikitina, Valentina E.

    2012-03-01

    The influence of bacterial lectin on photochemically induced fat cell lipolysis was studied. Resulting capsules were tested for ICG absorption by optical spectra measurements. To separate released and encapsulated ICG supernatant was removed and capsules were redispered in pure deionized water. Supernatant and capsule suspension spectra were measured separately. It was also found that pretreatment of tissue by lectin leads to acceleration of lipolysis at photochemical treatment. The data obtained can be used to enhance efficiency of photochemical therapy.

  8. Human Mannose-Binding Lectin Inhibitor Prevents Myocardial Injury and Arterial Thrombogenesis in a Novel Animal Model

    OpenAIRE

    Pavlov, Vasile I.; Tan, Ying S.; Erin E McClure; La Bonte, Laura R.; Zou, Chenhui; Gorsuch, William B.; Stahl, Gregory L.

    2015-01-01

    Myocardial infarction and coagulation disorders are leading causes of disability and death in the world. An important role of the lectin complement pathway in myocardial infarction and coagulation has been demonstrated in mice genetically deficient in lectin complement pathway proteins. However, these studies are limited to comparisons between wild-type and deficient mice and lack the ability to examine reversal/inhibition of injury after disease establishment. We developed a novel mouse that...

  9. Protection of gerbils from amebic liver abscess by immunization with the galactose-specific adherence lectin of Entamoeba histolytica.

    OpenAIRE

    Petri, W A; Ravdin, J I

    1991-01-01

    No protective antigens from Entamoeba histolytica have been previously defined. We tested the ability of the galactose-specific adherence lectin of E. histolytica to elicit a protective immune response in conjunction with Freund's incomplete and complete adjuvants. The gerbil (Meriones unguiculatus) model of an experimental amebic liver abscess was used. Gerbils were immunized intraperitoneally or subcutaneously with 10 micrograms of the affinity-purified lectin in complete Freund's adjuvant ...

  10. Specificity of lectin-immobilized fluorescent nanospheres for colorectal tumors in a mouse model which more resembles the clinical disease

    OpenAIRE

    Kitamura, Tokio; Sakuma, Shinji; Shimosato, Moe; Higashino, Haruki; Masaoka, Yoshie; Kataoka, Makoto; Yamashita, Shinji; Hiwatari, Ken-Ichiro; Kumagai, Hironori; Morimoto, Naoki; Koike, Seiji; Tobita, Etsuo; Hoffman, Robert M.; John C Gore; Pham, Wellington

    2014-01-01

    We are investigating an imaging agent that enables real-time and accurate diagnosis of early colorectal cancer at the intestinal mucosa by colonoscopy. The imaging agent is peanut agglutinin-immobilized polystyrene nanospheres with surface poly(N-vinylacetamide) chains encapsulating coumarin 6. Intracolonically-administered lectin-immobilized fluorescent nanospheres detect tumor-derived changes through molecular recognition of lectin for the terminal sugar of cancer-specific antigens on the m...

  11. Practical class for Biochemistry discipline addressing the influence of thermal processes in the activity of lectins from leguminous plants

    OpenAIRE

    Aline Matte; Patrícia Idalina de Lemos Rodrigues; Daniela Pereira Stocher; Renata Wadenphul de Moraes; Ludymila Schulz Barroso; Priscila Silva Corrêa; Cristiane Matte

    2014-01-01

    Lectins are proteins highly expressed in leguminous plants, which are capable of specifically recognize and bind carbohydrates, including those found in the erythrocyte membrane. The process of cooking food rich in lectins, such as beans, peas and lentils; promote protein denaturation, losing the native structure, and consequently, the ability of carbohydrate binding. This study presents the development of a protocol for practice classes of Biochemistry, permitting an easy way to verify meani...

  12. A 160-kilodalton epithelial cell surface glycoprotein recognized by plant lectins that inhibit the adherence of Actinomyces naeslundii.

    OpenAIRE

    Brennan, M J; Cisar, J O; Sandberg, A L

    1986-01-01

    The adherence of Actinomyces naeslundii to human epithelial (KB) cells is mediated by the interaction of a fimbrial lectin on this oral bacterium with epithelial cell receptors exposed by sialidase. The D-galactose- and N-acetyl-D-galactosamine-reactive plant lectins from peanut and from Bauhinia purpurea inhibit this interaction. This report describes the partial purification and characterization of a 160-kilodalton (kDa) cell surface glycoprotein which is the principal receptor for these le...

  13. High levels of serum mannose-binding lectin are associated with the severity of clinical signs of leptospirosis

    OpenAIRE

    K.A. Miranda; L.R.S. Vasconcelos; L.C.B.B. Coelho; J.L. Lima Filho; M. S. M. Cavalcanti; Moura, P.

    2009-01-01

    The clinical heterogeneity observed in leptospirosis may be associated with host factors or bacteria virulence. Human serum mannose-binding lectin (MBL) recognizes many pathogens, and low levels of this lectin are associated with susceptibility to infection. MBL is also implicated in the modulation of the inflammatory process. We determined the levels of serum MBL during leptospirosis infection. A double-antibody sandwich ELISA was used to detect the immunoreactive serum MBL. The ELISA plates...

  14. Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysis

    Institute of Scientific and Technical Information of China (English)

    Yu-Chieh Wang; Trevor R Leonardo; Ying Liu; Suzanne E Peterson; Louise C Laurent; Shinya Yamanaka; Jeanne F Loring; Masato Nakagawa; Ibon Garitaonandia; Ileana Slavin; Gulsah Altun; Robert M Lacharite; Kristopher L Nazor; Ha T Tran; Candace L Lynch

    2011-01-01

    Rapid and dependable methods for isolating human pluripotent stem cell (hPSC) populations are urgently needed for quality control in basic research and in cell-based therapy applications.Using lectin arrays,we analyzed glycoproteins extracted from 26 hPSC samples and 22 differentiated cell samples,and identified a small group of iectins with distinctive binding signatures that were sufficient to distinguish hPSCs from a variety of non-pluripotent cell types.These specific biomarkers were shared by all the 12 human embryonic stem cell and the 14 human induced pluripotent stem cell samples examined,regardless of the laboratory of origin,the culture conditions,the somatic cell type reprogrammed,or the reprogramming method used.We demonstrated a practical application of specific lectin binding by detecting hPSCs within a differentiated cell population with lectin-mediated staining followed by fluorescence microscopy and flow cytometry,and by enriching and purging viable hPSCs from mixed cell populations using lectin-mediated cell separation.Global gene expression analysis showed pluripotency-associated differential expression of specific fucosyltransferases and sialyltransferases,which may underlie these differences in protein glycosylation and lectin binding.Taken together,our results show that protein glycosylation differs considerably between pluripotent and non-pluripotent cells,and demonstrate that lectins may be used as biomarkers to monitor pluripotency in stem cell populations and for removal of viable hPSCs from mixed cell populations.

  15. Molecular Cloning of Lectin Gene from Lycoris radiata%石蒜凝集素基因的克隆

    Institute of Scientific and Technical Information of China (English)

    姚剑虹; 孙小芬; 唐克轩

    2002-01-01

    用石蒜科植物凝集素基因的保守序列为引物,利用RACE-PCR技术从石蒜幼嫩叶片中克隆出石蒜凝集素的全长cDNA.通过比较石蒜同其他石蒜科植物凝集素基因序列和推测的氨基酸序列,发现石蒜凝集素基因编码一具有信号肽的前体蛋白.石蒜凝集素同其他石蒜科植物凝集素一样为具有3个甘露糖专一结合盒的凝集素.%By the use of RACE-PCR technique with RNA extracted from Lycoris radiata young leaves as template andprimers specifically designed according to conservative sequences of Amaryllidaceae lectins, the full-length cDNA of Ly-coris radiata lectin was cloned. Through comparative analysis of Lycoris radiata lectin gene and its deduced amino acidsequence with those of other Amaryllidaceae species,it was found that Lycoris radiata lectin gene encoded a precursorlectin with signal peptide. Lycoris radiata lectin was a mannose-binding lectin with three mannose-binding boxes likelectins from other Amaryllidaceae species.

  16. Lectin-functionalized poly(glycidyl methacrylate)-block-poly(vinyldimethyl azlactone) surface supports for high avidity microbial capture

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, Ryan R [ORNL; Hinestrosa Salazar, Juan P [ORNL; Shubert, Katherine R [ORNL; Morrell, Jennifer L. [University of Tennessee, Knoxville (UTK); Pelletier, Dale A [ORNL; Messman, Jamie M [ORNL; Kilbey, II, S Michael [ORNL; Lokitz, Bradley S [ORNL; Retterer, Scott T [ORNL

    2013-01-01

    Microbial exopolysaccharides (EPS) play a critical and dynamic role in shaping the interactions between microbial community members and their local environment. The capture of targeted microbes using surface immobilized lectins that recognize specific extracellular oligosaccharide moieties offers a non-destructive method for functional characterization based on EPS content. In this report, we evaluate the use of the block co-polymer, poly(glycidyl methacrylate)-block-4,4-dimethyl-2-vinylazlactone (PGMA-b-PVDMA), as a surface support for lectin-specific microbial capture. Arrays of circular polymer supports ten micron in diameter were generated on silicon substrates to provide discrete, covalent coupling sites for Triticum vulgare and Lens culinaris lectins. These supports promoted microbe adhesion and colony formation in a lectin-specific manner. Silicon posts with similar topography containing only physisorbed lectins showed significantly less activity. These results demonstrate that micropatterned PGMA-b-PVDMA supports provide a unique platform for microbial capture and screening based on EPS content by combining high avidity lectin surfaces with three-dimensional topography.

  17. The effect of a lectin from the venom of the snake, Bothrops jararacussu, on tumor cell proliferation.

    Science.gov (United States)

    Pereira-Bittencourt, M; Carvalho, D D; Gagliardi, A R; Collins, D C

    1999-01-01

    Lectins have been used extensively as histochemical probes to describe changes in tumor cell surface and are known to influence the growth of cancer cells. In this study, we determined the effect of a lectin from the venom of Bothrops jararacussu (BJcuL) on the proliferation of a number of established human cancer cell lines. The growth of eight cancer cell lines was inhibited in a dose-related manner in the presence of BJcuL lectin. This lectin was most potent as an inhibitor of growth in renal (Caki-1 and A-498) and pancreatic (CFPAC-1) cancer cell lines with 50% inhibitory concentrations (IC50) of 1-2 mM. Melanoma (Wm115) and prostate (PC-3) cancer cells showed IC50 values of 7.9 and 8.5 mM, respectively, in the presence of BjcuL lectin whereas colon (Caco-2) and breast (MCF7) cancer cell lines showed no effect. Our results suggest that BJcuL lectin is an effective inhibitor of cell growth in some cancer cell lines. PMID:10628348

  18. Activation of endogenous C-type retroviral genomes by internal alpha-irradiation of mice with 224Radium

    International Nuclear Information System (INIS)

    Sensitive cocultivation techniques were applied to study the radiation-induced activation of endogenous retroviral genomes in different mouse strains by the alpha-emitting radionuclide 224Radium. Activated infectious C-type retroviruses were detected in spleen, bone marrow and bone tissues of C57BL/6-, BALB/c- and NMRI mice. The titres of high-dose-irradiated animals were higher than those found in low-dose-irradiated animals. Infectious retrovirus could be detected with a dose of 13.2 rad (maximum dose rate 0.9 rad/day) in the skeleton, and a dose of 4.2 rad (maximum dose rate 0.3 rad/day) in the spleen. The virus activation pattern was different in the three mouse strains. These data indicate that activation of endogenous retroviral genomes by alpha-irradiation shows a dose-effect relationship and a dependence on the genetic background of the mouse. (orig.)

  19. Molecular Modeling of Lectin-Like Protein from Acacia farnesiana Reveals a Possible Anti-Inflammatory Mechanism in Carrageenan-Induced Inflammation

    OpenAIRE

    Vanessa Erika Ferreira Abrantes; Bruno Anderson Matias da Rocha; Raphael Batista da Nóbrega; José Caetano Silva-Filho; Claudener Souza Teixeira; Benildo Sousa Cavada; Carlos Alberto de Almeida Gadelha; Sergio Henrique Ferreira; Jozi Godoy Figueiredo; Tatiane Santi-Gadelha; Plinio Delatorre

    2013-01-01

    Acacia farnesiana lectin-like protein (AFAL) is a chitin-binding protein and has been classified as phytohaemagglutinin from Phaseolus vulgaris (PHA). Legume lectins are examples for structural studies, and this family of proteins shows a remarkable conservation in primary, secondary, and tertiary structures. Lectins have ability to reduce the effects of inflammation caused by phlogistic agents, such as carrageenan (CGN). This paper explains the anti-inflammatory activity of AFAL through stru...

  20. Mannose-binding lectin and maladies of the bowel and liver

    Institute of Scientific and Technical Information of China (English)

    Daniel L Worthley; Peter G Bardy; David L Gordon; Charles G Mullighan

    2006-01-01

    Mannose-binding lectin (MBL) is a pattern-recognition molecule that binds to characteristic carbohydrate motifs present on the surface of many different pathogens.MBL binding stimulates the immune system via the lectin pathway of complement activation. In certain clinical situations, often characterized by pre-existing immune compromise, MBL deficiency increases the risk of infectious and other disease-specific complications. Many of the key pathogenic processes inherent to common gastroenterological diseases, such as infection, immunological damage, and carcinogenesis, have been linked to MBL. This editorial reviews the biology of MBL, outlines key disease associations to document the breadth of influence of MBL, and finally, highlights the relevance of MBL to both gastroenterological health and disease.