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Sample records for calpain inhibitor ak

  1. Calpain inhibitor nanocrystals prepared using Nano Spray Dryer B-90

    Science.gov (United States)

    Baba, Koichi; Nishida, Kohji

    2012-08-01

    The Nano Spray Dryer B-90 offers a new, simple, and alternative approach for the production of drug nanocrystals. Among attractive drugs, calpain inhibitor that inhibits programmed cell death `apoptosis' is a candidate for curing apoptosis-mediated intractable diseases such as Alzheimer's disease and Parkinson's disease. In this study, the preparation of calpain inhibitor nanocrystals using Nano Spray Dryer B-90 was demonstrated. The particle sizes were controlled by means of selecting mesh aperture sizes. The obtained average particle sizes were in the range of around 300 nm to submicron meter.

  2. Calpain inhibitors reduce retinal hypoxia in ischemic retinopathy by improving neovascular architecture and functional perfusion.

    Science.gov (United States)

    Hoang, Mien V; Smith, Lois E H; Senger, Donald R

    2011-04-01

    In ischemic retinopathies, underlying hypoxia drives abnormal neovascularization that damages retina and causes blindness. The abnormal neovasculature is tortuous and leaky and fails to alleviate hypoxia, resulting in more pathological neovascularization and retinal damage. With an established model of ischemic retinopathy we found that calpain inhibitors, when administered in moderation, reduced architectural abnormalities, reduced vascular leakage, and most importantly reduced retinal hypoxia. Mechanistically, these calpain inhibitors improved stability and organization of the actin cytoskeleton in retinal endothelial cells undergoing capillary morphogenesis in vitro, and they similarly improved organization of actin cables within new blood vessels in vivo. Hypoxia induced calpain activity in retinal endothelial cells and severely disrupted the actin cytoskeleton, whereas calpain inhibitors preserved actin cables under hypoxic conditions. Collectively, these findings support the hypothesis that hyper-activation of calpains by hypoxia contributes to disruption of the retinal endothelial cell cytoskeleton, resulting in formation of neovessels that are defective both architecturally and functionally. Modest suppression of calpain activity with calpain inhibitors restores cytoskeletal architecture and promotes formation of a functional neovasculature, thereby reducing underlying hypoxia. In sharp contrast to "anti-angiogenesis" strategies that cannot restore normoxia and may aggravate hypoxia, the therapeutic strategy described here does not inhibit neovascularization. Instead, by improving the function of neovascularization to reduce underlying hypoxia, moderate calpain inhibition offers a method for alleviating retinal ischemia, thereby suggesting a new treatment paradigm based on improvement rather than inhibition of new blood vessel growth. Copyright © 2010 Elsevier B.V. All rights reserved.

  3. Mechanism of Action of Thalassospiramides, A New Class of Calpain Inhibitors

    KAUST Repository

    Lu, Liang

    2015-03-05

    Thalassospiramides comprise a large family of lipopeptide natural products produced by Thalassospira and Tistrella marine bacteria. Here we provide further evidence of their nanomolar inhibitory activity against the human calpain 1 protease. Analysis of structure-activity relationship data supported our hypothesis that the rigid 12-membered ring containing an α,β-unsaturated carbonyl moiety is the pharmacologically active functional group, in contrast to classic electrophilic "warheads" in known calpain inhibitors. Using a combination of chemical modifications, mass spectrometric techniques, site-directed mutagenesis, and molecular modeling, we show the covalent binding of thalassospiramide\\'s α,β-unsaturated carbonyl moiety to the thiol group of calpain\\'s catalytic Cys115 residue by a Michael 1,4-addition reaction. As nanomolar calpain inhibitors with promising selectivity and low toxicity from natural sources are rare, we consider thalassospiramides as promising drug leads.

  4. Therapeutic Efficacy of E-64-d, a Selective Calpain Inhibitor, in Experimental Acute Spinal Cord Injury

    Directory of Open Access Journals (Sweden)

    Zifeng Zhang

    2015-01-01

    Full Text Available This study aims to investigate the therapeutic effect of calpain inhibitor E-64-d on SCI and to find a new approach to treat SCI. When an SCI rat model was established, it was immediately administered with E-64-d. RT-PCR and Western blotting were used to determine the protein and mRNA levels of calpain 1 and 68-kD NFP. TUNEL staining and NeuN labeling were performed to analyze neuronal apoptosis in the lesion. Immunohistochemistry assay was carried out to observe the expressions of calpain 1 and GFAP. Cyclooxygenase-2 activity was measured to show the immune response status. Locomotor function was evaluated by inclined plane test and Basso, Beattie, and Bresnahan locomotor rating scale. The results showed that calpain 1 was activated after SCI occurred. Treatment with E-64-d decreased expressions of calpain 1 and GFAP, alleviated neuronal apoptosis, inhibited cyclooxygenase-2 activity, and resulted in the promoted locomotor function. Furthermore, combination of E-64-d and MP had better efficacy than did E-64-d or MP alone. E-64-d is expected to be applied to treat SCI, and its alliance with MP may provide a valid strategy for SCI therapy.

  5. In Silico Affinity Profiling of Neuroactive Polyphenols for Post-Traumatic Calpain Inactivation: A Molecular Docking and Atomistic Simulation Sensitivity Analysis

    Directory of Open Access Journals (Sweden)

    Pradeep Kumar

    2014-12-01

    Full Text Available Calcium-activated nonlysosomal neutral proteases, calpains, are believed to be early mediators of neuronal damage associated with neuron death and axonal degeneration after traumatic neural injuries. In this study, a library of biologically active small molecular weight calpain inhibitors was used for model validation and inhibition site recognition. Subsequently, two natural neuroactive polyphenols, curcumin and quercetin, were tested for their sensitivity and activity towards calpain’s proteolytic sequence and compared with the known calpain inhibitors via detailed molecular mechanics (MM, molecular dynamics (MD, and docking simulations. The MM and MD energy profiles (SJA6017 < AK275 < AK295 < PD151746 < quercetin < leupeptin < PD150606 < curcumin < ALLN < ALLM < MDL-28170 < calpeptin and the docking analysis (AK275 < AK295 < PD151746 < ALLN < PD150606 < curcumin < leupeptin < quercetin < calpeptin < SJA6017 < MDL-28170 < ALLM demonstrated that polyphenols conferred comparable calpain inhibition profiling. The modeling paradigm used in this study provides the first detailed account of corroboration of enzyme inhibition efficacy of calpain inhibitors and the respective calpain–calpain inhibitor molecular complexes’ energetic landscape and in addition stimulates the polyphenol bioactive paradigm for post-SCI intervention with implications reaching to experimental in vitro, in cyto, and in vivo studies.

  6. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 O08529|CAN2_MOUSE Calpain-2 catalytic subunit precursor (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain...) (80 kDa M-calpain subunit) (CALP80) - Mus musculus (Mouse) 0 ...

  7. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 P17655|CAN2_HUMAN Calpain-2 catalytic subunit precursor (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain...) (Calpain large polypeptide L2) - Homo sapiens (Human) 0 ...

  8. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (O08529) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (80 kDa M-calpain subunit) (CALP80) CAN2_MOUSE 5e-53 ...

  9. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (P17655) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Calpain large polypeptide L2) CAN2_HUMAN 2e-53 ...

  10. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (O08529) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (80 kDa M-calpain subunit) (CALP80) CAN2_MOUSE 1e-38 ...

  11. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (P17655) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Calpain large polypeptide L2) CAN2_HUMAN 6e-40 ...

  12. Calpain inhibitors and antioxidants act synergistically to prevent cell necrosis: effects of the novel dual inhibitors (cysteine protease inhibitor and antioxidant) BN 82204 and its pro-drug BN 82270.

    Science.gov (United States)

    Pignol, Bernadette; Auvin, Serge; Carré, Denis; Marin, Jean-Grégoire; Chabrier, Pierre-Etienne

    2006-08-01

    Cell death is a common feature observed in neurodegenerative disorders, and is often associated with calpain activation and overproduction of reactive oxygen species (ROS). This study investigated the use of calpain inhibitors and antioxidants in combination to protect cells against necrosis. Maitotoxin (MTX), which induces a massive influx of calcium, was used to provoke neuronal cell death. This toxin increased, in a concentration-dependent manner, both calpain activity and ROS formation. Calpain inhibitors or antioxidants inhibited MTX-induced necrosis only marginally (below 20%), whereas their association protected against cell death by 40-66% in a synergistic manner. BN 82204, which possesses both calpain-cathepsin L inhibitory and antioxidant properties, and its acetylated pro-drug BN 82270, totally protected cells at 100 microm. The pro-drug BN 82270, which had better cell penetration, was twice as effective as the active principle BN 82204 in protecting glioma C6 or neuroblastoma SHSY5Y cells against death. These results suggest the potential therapeutic relevance of using a single molecule with multiple activities (cysteine protease inhibitor/antioxidant), and warrant further in vivo investigations in models of neuronal disorders.

  13. Effects of a Caspase and a Calpain Inhibitor on Resting Energy Expenditures in Normal and Hypermetabolic Rats: a Pilot Study

    Science.gov (United States)

    VANA, P. G.; LAPORTE, H. M.; KENNEDY, R. H.; GAMELLI, R. L.; MAJETSCHAK, M.

    2017-01-01

    Summary Several diseases induce hypermetabolism, which is characterized by increases in resting energy expenditures (REE) and whole body protein loss. Exaggerated protein degradation is thought to be the driving force underlying this response. The effects of caspase and calpain inhibitors on REE in physiological and hypermetabolic conditions, however, are unknown. Thus, we studied whether MDL28170 (calpain inhibitor) or z-VAD-fmk (caspase inhibitor) affect REE under physiological conditions and during hypermetabolism post-burn. Rats were treated five times weekly and observed for 6 weeks. Treatment was started 2 h (early) or 48 h (late) after burn. In normal rats, MDL28170 transiently increased REE to 130 % of normal during week 2–4. z-VAD-fmk reduced REE by 20–25 % throughout the observation period. Within 14 days after burns, REE increased to 130±5 %. Whereas MDL28170/early treatment did not affect REE, MDL28170/late transiently increased REE to 180±10 % of normal by week 4 post-burn. In contrast, with z-VAD-fmk/early REE remained between 90–110 % of normal post-burn. z-VAD-fmk/late did not affect burn-induced increases in REE. These data suggest that caspase cascades contribute to the development of hypermetabolism and that burn-induced hypermetabolism can be pharmacologically modulated. Our data point towards caspase cascades as possible therapeutic targets to attenuate hypermetabolism after burns, and possibly in other catabolic disease processes. PMID:27070748

  14. Arabidopsis CDS blastp result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 At1g55350.4 calpain-type cysteine protease family identical to calpain...-like protein GI:20268660 from [Arabidopsis thaliana]; contains Pfam profiles: PF00648 Calpain family... cysteine protease, PF01067 Calpain large subunit,domain III; identical to cDNA calpain-like protein GI:20268659 1e-150 ...

  15. Arabidopsis CDS blastp result: AK121261 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121261 J023104H13 At1g55350.4 calpain-type cysteine protease family identical to calpain...-like protein GI:20268660 from [Arabidopsis thaliana]; contains Pfam profiles: PF00648 Calpain family... cysteine protease, PF01067 Calpain large subunit,domain III; identical to cDNA calpain-like protein GI:20268659 0.0 ...

  16. Susceptibility of Phytomonas serpens to calpain inhibitors in vitro: interference on the proliferation, ultrastructure, cysteine peptidase expression and interaction with the invertebrate host

    Science.gov (United States)

    de Oliveira, Simone Santiago Carvalho; Gonçalves, Diego de Souza; Garcia-Gomes, Aline dos Santos; Gonçalves, Inês Correa; Seabra, Sergio Henrique; Menna-Barreto, Rubem Figueiredo; Lopes, Angela Hampshire de Carvalho Santos; D’Avila-Levy, Claudia Masini; dos Santos, André Luis Souza; Branquinha, Marta Helena

    2016-01-01

    A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites’ physiology. PMID:27925020

  17. Neuroprotection of vestibular sensory cells from gentamicin ototoxicity obtained using nitric oxide synthase inhibitors, reactive oxygen species scavengers, brain-derived neurotrophic factors and calpain inhibitors.

    Science.gov (United States)

    Takumida, Masaya; Anniko, Matti; Shimizu, Akira; Watanabe, Hiroshi

    2003-01-01

    In order to devise a new treatment for inner ear disorders, the efficacy of a nitric oxide synthase inhibitor (L-N(G)-nitroarginine methylester [L-NAME]), a radical scavenger (D-methionine), a neurotrophin (brain-derived neurotrophic factor [BDNF]) and a calpain inhibitor (leupeptin) for protection from hair cell damage was investigated. The effects of these drugs on gentamicin-induced production of nitric oxide (NO) and reactive oxygen species (ROS) were studied by means of the fluorescence indicators 4,5-diaminofluorescein diacetate and dihydrotetramethylrosamine. The effect on gentamicin-induced vestibular hair cell damage was examined by using an in vitro LIVE/DEAD system. L-NAME inhibited the production of NO, D-methionine and BDNF restricted the production of ROS and leupeptin inhibited neither NO nor ROS. All the drugs used limited the vestibular hair cell damage caused by gentamicin. The combinations L-NAME + BDNF, L-NAME + leupeptin and D-methionine + BDNF had a significantly stronger preventive effect on hair cell damage. It is suggested that combined treatment with a radical inhibitor and either a neurotrophin or calpain inhibitor may help to treat inner ear disorders more effectively.

  18. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q07009) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_RAT 9e-52 ...

  19. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q92178) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_CHICK 3e-38 ...

  20. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 Q07009|CAN2_RAT Calpain-2 catalytic subunit precursor (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) - Rattus norvegicus (Rat) 0 ...

  1. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (P06814) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Fragment) CAN2_RABIT 6e-16 ...

  2. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 P06814|CAN2_RABIT Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Fragment) - Oryctolagus cuniculus (Rabbit) 0 ...

  3. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 Q92178|CAN2_CHICK Calpain-2 catalytic subunit precursor (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) - Gallus gallus (Chicken) 0 ...

  4. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q07009) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_RAT 4e-38 ...

  5. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 P43367|CAN2_PIG Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Fragment) - Sus scrofa (Pig) 0 ...

  6. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q9GLG1) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_MACFA 3e-53 ...

  7. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (Q92178) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_CHICK 1e-11 ...

  8. UniProt search blastx result: AK287903 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287903 J065211G19 Q9GLG1|CAN2_MACFA Calpain-2 catalytic subunit precursor (EC 3.4.22.53) (Calpain...-2 large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain

  9. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q92178) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2... large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_CHICK 1e-51 ...

  10. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q9GLG1) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_MACFA 8e-40 ...

  11. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (Q92178) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) CAN2_CHICK 3e-11 ...

  12. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (P06814) Calpain-2 catalytic subunit (EC 3.4.22.53) (Calpain-2 ...large subunit) (Calcium-activated neutral proteinase 2) (CANP 2) (Calpain M-type) (M-calpain) (Millimolar-calpain) (Fragment) CAN2_RABIT 2e-16 ...

  13. A cardioprotective agent of a novel calpain inhibitor, SNJ-1945, exerts beta1 actions on left ventricular mechanical work and energetics.

    Science.gov (United States)

    Yoshikawa, Yoshiro; Zhang, Guo-Xing; Obata, Koji; Matsuyoshi, Hiroko; Asada, Keiji; Taniguchi, Shigeki; Takaki, Miyako

    2010-08-01

    We have previously shown that a newly developed calpain inhibitor, SNJ-1945 (SNJ), with good aqueous solubility prevents the heart from KCl arrest-reperfusion injury associated with the impairment of total Ca(2+) handling by inhibiting the proteolysis of alpha-fodrin as a cardioplegia. The aim of the present study was to investigate certain actions of this calpain inhibitor, SNJ, on left ventricular (LV) mechanical work and energetics in cross-circulated excised rat hearts undergoing blood perfusion with 40 microM SNJ. Mean end-systolic pressure at midrange LV volume and systolic pressure-volume area (PVA) at mLVV (a total mechanical energy/beat) were significantly increased by SNJ perfusion (P work and energetics mediated via beta(1)-adrenergic receptors associated with the enhancement of total Ca(2+) handling in excitation-contraction coupling and with unchanged contractile efficiency. In clinical settings, this pharmacological action of SNJ is beneficial as an additive agent for cardioplegia.

  14. [Analgesic effect of calpain inhibitor ALLN on the zymosan-induced paw inflammatory pain and its effect on the expression of cyclooxygenase-2 in the spinal dorsal horn].

    Science.gov (United States)

    Wang, Jing-Jie; Chen, Guang-Jun; Chen, Wen; Du, Jin; Luo, Ai-Lun; Huang, Yu-Guang

    2012-02-01

    To examine the analgesic effect of calpain inhibitor ALLN on the zymosan-induced paw inflammatory pain and its effect on the expression of cyclooxygenase-2 (COX-2) in the spinal dorsal horn. Forty-eight Sprague-Dawley rats were equally divided into three groups: control group, sham-operated group, and zymosan group. According to Meller's method, zymosan (1.25 mg) was injected intraplantarly to induce paw inflammation in zymosan group; an equal volume of PBS was administered in the sham-operated group. Mechanical withdrawal threshold (MWT) and maximum thickness of paw were tested or measured before and 0.5, 1, 2, 4, 8, and 24 hours after injection. All rats were killed at different occasions following surgery to examine calpain activity in the spinal dorsal horn with Western blot analysis. Another sixty-four Sprague-Dawley rats were divided into three groups: sham-operated group, zymosan-induced paw inflammation with intraperitoneal dimethyl sulphoxide (DMSO) treatment group, and zymosan-induced paw inflammation with intraperitoneal calpain inhibitor ALLN treatment group. MWT and maximum thickness of paw were tested or measured before and 0.5, 1, 2, 4, 8, and 24 hours after injection. All rats were killed at different occasions following surgery to examine the COX-2 expression in the spinal dorsal horn with Western blot analysis. MWT significantly decreased in the rats with zymosan-induced paw inflammation, while the maximum thickness of paw significantly increased, compared with control and sham-operated rats (P horn was dramatically activated after zymosan injection (P horn compared with DMSO treatment (P effective to attenuate zymosan-induced paw inflammatory pain. Calpain activation may be one aspect of the signaling cascade that increases the COX-2 expression in the spinal cord and contributes to mechanical hyperalgesia after peripheral inflammatory injury.

  15. Inhibitors of cysteine cathepsin and calpain do not prevent ultraviolet-B-induced apoptosis in human keratinocytes and HeLa cells

    DEFF Research Database (Denmark)

    Bang, Bo; Baadsgaard, Ole; Skov, Lone

    2004-01-01

    been demonstrated to play a role in the execution of programmed cell death induced by other stimuli, e.g. TNF-alpha. The purpose of the present study was therefore to investigate whether inhibitors of cysteine cathepsins and calpains could prevent UVB-induced apoptosis in HeLa cells and keratinocytes....... This was done by investigating the effect of the irreversible cysteine protease inhibitor zFA-fmk, the cathepsin B inhibitor CA-074-Me and the calpain inhibitor ALLN on the viability of UVB-irradiated human keratinocytes and HeLa cells. At concentrations of 10 microM and above zVAD-fmk conferred partial dose......-dependent protection against UVB-induced apoptosis in HeLa cells and keratinocytes. Moreover, caspase-3 activity was completely blocked at zVAD-fmk concentrations of 1 microM in HeLa cells. This indicates that caspase-independent mechanisms could be involved in UVB-induced apoptosis. However, the protease inhibitors z...

  16. Arabidopsis CDS blastp result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 At1g55350.4 calpain-type cysteine protease family identical to calpain...-like protein GI:20268660 from [Arabidopsis thaliana]; contains Pfam profiles: PF00648 Calpain famil...y cysteine protease, PF01067 Calpain large subunit,domain III; identical to cDNA calpain-like protein GI:20268659 0.0 ...

  17. Chronic administration of a leupeptin-derived calpain inhibitor fails to ameliorate severe muscle pathology in a canine model of Duchenne muscular dystrophy

    Directory of Open Access Journals (Sweden)

    Martin K Childers

    2012-01-01

    Full Text Available Calpains likely play a role in the pathogenesis of Duchenne muscular dystrophy (DMD. Accordingly, calpain inhibition may provide therapeutic benefit to DMD patients. In the present study, we sought to measure benefit from administration of a novel calpain inhibitor, C101, in a canine muscular dystrophy model. Specifically, we tested the hypothesis that treatment with C101 mitigates progressive weakness and severe muscle pathology observed in young dogs with golden retriever muscular dystrophy (GRMD. Young (6 week-old GRMD dogs were treated daily with either C101 (17mg/kg twice daily oral dose, n=9 or placebo (vehicle only, n=7 for 8 weeks. A battery of functional tests, including tibiotarsal joint angle, muscle/fat composition, and pelvic limb muscle strength were performed at baseline and every two weeks during the 8-week study. Results indicate that C101-treated GRMD dogs maintained strength in their cranial pelvic limb muscles (tibiotarsal flexors while placebo-treated dogs progressively lost strength. However, concomitant improvement was not observed in posterior pelvic limb muscles (tibiotarsal extensors. C101 treatment did not mitigate force drop following repeated eccentric contractions and no improvement was seen in the development of joint contractures, lean muscle mass or muscle histopathology. Taken together, these data do not support the hypothesis that treatment with C101 mitigates progressive weakness or ameliorates severe muscle pathology observed in young dogs with GRMD.

  18. SwissProt search result: AK099458 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099458 J013022O12 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 4e-11 ...

  19. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (Q27970) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_BOVIN 2e-11 ...

  20. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q9GLG2) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MACFA 1e-39 ...

  1. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (O35350) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MOUSE 9e-41 ...

  2. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (O35350) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MOUSE 2e-53 ...

  3. SwissProt search result: AK099458 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099458 J013022O12 (P06815) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) (Fragment) CAN1_RABIT 2e-11 ...

  4. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 2e-12 ...

  5. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (P97571) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_RAT 2e-11 ...

  6. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q27970) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_BOVIN 5e-52 ...

  7. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q27970) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_BOVIN 2e-39 ...

  8. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (P97571) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_RAT 1e-53 ...

  9. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (P07384) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_HUMAN 2e-12 ...

  10. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 2e-12 ...

  11. SwissProt search result: AK065151 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065151 J013002B09 (P06815) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) (Fragment) CAN1_RABIT 2e-11 ...

  12. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (P97571) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_RAT 2e-11 ...

  13. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (O35350) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MOUSE 2e-11 ...

  14. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 7e-52 ...

  15. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (O35350) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MOUSE 2e-11 ...

  16. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q9GLG2) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MACFA 1e-52 ...

  17. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (Q9GLG2) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MACFA 2e-12 ...

  18. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (Q9GLG2) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_MACFA 2e-12 ...

  19. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (P06815) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) (Fragment) CAN1_RABIT 8e-12 ...

  20. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (P97571) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_RAT 7e-41 ...

  1. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (Q27970) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_BOVIN 2e-11 ...

  2. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (P07384) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_HUMAN 1e-39 ...

  3. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (P07384) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_HUMAN 2e-12 ...

  4. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (P07384) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_HUMAN 2e-52 ...

  5. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 4e-39 ...

  6. SwissProt search result: AK065151 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065151 J013002B09 (P35750) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1 ...large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) CAN1_PIG 4e-11 ...

  7. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (P06815) Calpain-1 catalytic subunit (EC 3.4.22.52) (Calpain-1... large subunit) (Calcium-activated neutral proteinase 1) (CANP 1) (Calpain mu-type) (muCANP) (Micromolar-calpain) (Fragment) CAN1_RABIT 6e-12 ...

  8. Identification of active Plasmodium falciparum calpain to establish screening system for Pf-calpain-based drug development

    Directory of Open Access Journals (Sweden)

    Soh Byoung

    2013-02-01

    Full Text Available Abstract Background With the increasing resistance of malaria parasites to available drugs, there is an urgent demand to develop new anti-malarial drugs. Calpain inhibitor, ALLN, is proposed to inhibit parasite proliferation by suppressing haemoglobin degradation. This provides Plasmodium calpain as a potential target for drug development. Pf-calpain, a cysteine protease of Plasmodium falciparum, belongs to calpain-7 family, which is an atypical calpain not harboring Ca2+-binding regulatory motifs. In this present study, in order to establish the screening system for Pf-calpain specific inhibitors, the active form of Pf-calpain was first identified. Methods Recombinant Pf-calpain including catalytic subdomain IIa (rPfcal-IIa was heterologously expressed and purified. Enzymatic activity was determined by both fluorogenic substrate assay and gelatin zymography. Molecular homology modeling was carried out to address the activation mode of Pf-calpain in the aspect of structural moiety. Results Based on the measurement of enzymatic activity and protease inhibitor assay, it was found that the active form of Pf-calpain only contains the catalytic subdomain IIa, suggesting that Pf-calpain may function as a monomeric form. The sequence prediction indicates that the catalytic subdomain IIa contains all amino acid residues necessary for catalytic triad (Cys-His-Asn formation. Molecular modeling suggests that the Pf-calpain subdomain IIa makes an active site, holding the catalytic triad residues in their appropriate orientation for catalysis. The mutation analysis further supports that those amino acid residues are functional and have enzymatic activity. Conclusion The identified active form of Pf-calpain could be utilized to establish high-throughput screening system for Pf-calpain inhibitors. Due to its unique monomeric structural property, Pf-calpain could be served as a novel anti-malarial drug target, which has a high specificity for malaria parasite

  9. SwissProt search result: AK072218 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072218 J013167O21 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pro...teinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 5e-45 ...

  10. SwissProt search result: AK103409 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103409 J033128E16 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pro...teinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 1e-12 ...

  11. SwissProt search result: AK065151 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065151 J013002B09 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pro...teinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 7e-12 ...

  12. SwissProt search result: AK064381 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064381 002-108-E01 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pr...oteinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 9e-60 ...

  13. SwissProt search result: AK099458 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099458 J013022O12 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pro...teinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 6e-12 ...

  14. SwissProt search result: AK059278 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059278 001-025-C08 (Q9VT65) Calpain B (EC 3.4.22.-) (Calcium-activated neutral pr...oteinase B) (CANP B) [Contains: Calpain B catalytic subunit 1; Calpain B catalytic subunit 2] CANB_DROME 4e-13 ...

  15. Calpain Activator Dibucaine Induces Platelet Apoptosis

    Directory of Open Access Journals (Sweden)

    Jun Liu

    2011-03-01

    Full Text Available Calcium-dependent calpains are a family of cysteine proteases that have been demonstrated to play key roles in both platelet glycoprotein Ibα shedding and platelet activation and altered calpain activity is associated with thrombotic thrombocytopenic purpura. Calpain activators induce apoptosis in several types of nucleated cells. However, it is not clear whether calpain activators induce platelet apoptosis. Here we show that the calpain activator dibucaine induced several platelet apoptotic events including depolarization of the mitochondrial inner transmembrane potential, up-regulation of Bax and Bak, down-regulation of Bcl-2 and Bcl-XL, caspase-3 activation and phosphatidylserine exposure. Platelet apoptosis elicited by dibucaine was not affected by the broad spectrum metalloproteinase inhibitor GM6001. Furthermore, dibucaine did not induce platelet activation as detected by P-selectin expression and PAC-1 binding. However, platelet aggregation induced by ristocetin or α-thrombin, platelet adhesion and spreading on von Willebrand factor were significantly inhibited in platelets treated with dibucaine. Taken together, these data indicate that dibucaine induces platelet apoptosis and platelet dysfunction.

  16. Inhibition of calpain blocks the phagosomal escape of Listeria monocytogenes.

    Directory of Open Access Journals (Sweden)

    Gloria Lopez-Castejon

    Full Text Available Listeria monocytogenes is a gram-positive facultative intracellular bacterium responsible for the food borne infection listeriosis, affecting principally the immunocompromised, the old, neonates and pregnant women. Following invasion L. monocytogenes escapes the phagosome and replicates in the cytoplasm. Phagosome escape is central to L. monocytogenes virulence and is required for initiating innate host-defence responses such as the secretion of the cytokine interleukin-1. Phagosome escape of L. monocytogenes is reported to depend upon host proteins such as γ-interferon-inducible lysosomal thiol reductase and the cystic fibrosis transmembrane conductance regulator. The host cytosolic cysteine protease calpain is required in the life cycle of numerous pathogens, and previous research reports an activation of calpain by L. monocytogenes infection. Thus we sought to determine whether host calpain was required for the virulence of L. monocytogenes. Treatment of macrophages with calpain inhibitors blocked escape of L. monocytogenes from the phagosome and consequently its proliferation within the cytosol. This was independent of any direct effect on the production of bacterial virulence factors or of a bactericidal effect. Furthermore, the secretion of interleukin-1β, a host cytokine whose secretion induced by L. monocytogenes depends upon phagosome escape, was also blocked by calpain inhibition. These data indicate that L. monocytogenes co-opts host calpain to facilitate its escape from the phagosome, and more generally, that calpain may represent a cellular Achilles heel exploited by pathogens.

  17. Inhibition of calpain blocks the phagosomal escape of Listeria monocytogenes.

    Science.gov (United States)

    Lopez-Castejon, Gloria; Corbett, David; Goldrick, Marie; Roberts, Ian S; Brough, David

    2012-01-01

    Listeria monocytogenes is a gram-positive facultative intracellular bacterium responsible for the food borne infection listeriosis, affecting principally the immunocompromised, the old, neonates and pregnant women. Following invasion L. monocytogenes escapes the phagosome and replicates in the cytoplasm. Phagosome escape is central to L. monocytogenes virulence and is required for initiating innate host-defence responses such as the secretion of the cytokine interleukin-1. Phagosome escape of L. monocytogenes is reported to depend upon host proteins such as γ-interferon-inducible lysosomal thiol reductase and the cystic fibrosis transmembrane conductance regulator. The host cytosolic cysteine protease calpain is required in the life cycle of numerous pathogens, and previous research reports an activation of calpain by L. monocytogenes infection. Thus we sought to determine whether host calpain was required for the virulence of L. monocytogenes. Treatment of macrophages with calpain inhibitors blocked escape of L. monocytogenes from the phagosome and consequently its proliferation within the cytosol. This was independent of any direct effect on the production of bacterial virulence factors or of a bactericidal effect. Furthermore, the secretion of interleukin-1β, a host cytokine whose secretion induced by L. monocytogenes depends upon phagosome escape, was also blocked by calpain inhibition. These data indicate that L. monocytogenes co-opts host calpain to facilitate its escape from the phagosome, and more generally, that calpain may represent a cellular Achilles heel exploited by pathogens.

  18. Contribution of Calpain and Caspases to Cell Death in Cultured Monkey RPE Cells.

    Science.gov (United States)

    Nakajima, Emi; Hammond, Katherine B; Hirata, Masayuki; Shearer, Thomas R; Azuma, Mitsuyoshi

    2017-10-01

    AMD is the leading cause of human vision loss after 65 years of age. Several mechanisms have been proposed: (1) age-related failure of the choroidal vasculature leads to loss of RPE; (2) RPE dysfunctions due to accumulation of phagocytized, but unreleased A2E (N-retinylidene-N-retinylethanolamine); (3) zinc deficiency activation of calpain and caspase proteases, leading to cell death. The purpose of the present study is to compare activation of calpain and caspase in monkey RPE cells cultured under hypoxia or with A2E. Monkey primary RPE cells were cultured under hypoxic conditions in a Gaspak pouch or cultured with synthetic A2E. Immunoblotting was used to detect activation of calpain and caspase. Calpain inhibitor, SNJ-1945, and pan-caspase inhibitor, z-VAD-fmk, were used to confirm activation of the proteases. (1) Hypoxia and A2E each decreased viability of RPE cells in a time-dependent manner. (2) Incubation under hypoxia alone induced activation of calpain, but not caspases. SNJ-1945 inhibited calpain activation, but z-VAD-fmk did not. (3) Incubation with A2E alone induced activation of calpain, caspase-9, and caspase-3. SNJ-1945 inhibited calpain activation. z-VAD-fmk inhibited caspase activation, suggesting no interaction between calpain and caspases. Hypoxia activated the calpain pathway, while A2E activated both calpain and caspase pathways in monkey RPE cells. Such knowledge may be utilized in the treatment of AMD if inhibitor drugs against calpain and/or caspase are used to prevent RPE dysfunction caused by hypoxia or A2E.

  19. Nitric oxide inhibits calpain-mediated proteolysis of talin in skeletal muscle cells

    Science.gov (United States)

    Koh, T. J.; Tidball, J. G.

    2000-01-01

    We tested the hypothesis that nitric oxide can inhibit cytoskeletal breakdown in skeletal muscle cells by inhibiting calpain cleavage of talin. The nitric oxide donor sodium nitroprusside prevented many of the effects of calcium ionophore on C(2)C(12) muscle cells, including preventing talin proteolysis and release into the cytosol and reducing loss of vinculin, cell detachment, and loss of cellular protein. These results indicate that nitric oxide inhibition of calpain protected the cells from ionophore-induced proteolysis. Calpain inhibitor I and a cell-permeable calpastatin peptide also protected the cells from proteolysis, confirming that ionophore-induced proteolysis was primarily calpain mediated. The activity of m-calpain in a casein zymogram was inhibited by sodium nitroprusside, and this inhibition was reversed by dithiothreitol. Previous incubation with the active site-targeted calpain inhibitor I prevented most of the sodium nitroprusside-induced inhibition of m-calpain activity. These data suggest that nitric oxide inhibited m-calpain activity via S-nitrosylation of the active site cysteine. The results of this study indicate that nitric oxide produced endogenously by skeletal muscle and other cell types has the potential to inhibit m-calpain activity and cytoskeletal proteolysis.

  20. Moderation of calpain activity promotes neovascular integration and lumen formation during VEGF-induced pathological angiogenesis.

    Directory of Open Access Journals (Sweden)

    Mien V Hoang

    2010-10-01

    Full Text Available Successful neovascularization requires that sprouting endothelial cells (ECs integrate to form new vascular networks. However, architecturally defective, poorly integrated vessels with blind ends are typical of pathological angiogenesis induced by vascular endothelial growth factor-A (VEGF, thereby limiting the utility of VEGF for therapeutic angiogenesis and aggravating ischemia-related pathologies. Here we investigated the possibility that over-exuberant calpain activity is responsible for aberrant VEGF neovessel architecture and integration. Calpains are a family of intracellular calcium-dependent, non-lysosomal cysteine proteases that regulate cellular functions through proteolysis of numerous substrates.In a mouse skin model of VEGF-driven angiogenesis, retroviral transduction with dominant-negative (DN calpain-I promoted neovessel integration and lumen formation, reduced blind ends, and improved vascular perfusion. Moderate doses of calpain inhibitor-I improved VEGF-driven angiogenesis similarly to DN calpain-I. Conversely, retroviral transduction with wild-type (WT calpain-I abolished neovessel integration and lumen formation. In vitro, moderate suppression of calpain activity with DN calpain-I or calpain inhibitor-I increased the microtubule-stabilizing protein tau in endothelial cells (ECs, increased the average length of microtubules, increased actin cable length, and increased the interconnectivity of vascular cords. Conversely, WT calpain-I diminished tau, collapsed microtubules, disrupted actin cables, and inhibited integration of cord networks. Consistent with the critical importance of microtubules for vascular network integration, the microtubule-stabilizing agent taxol supported vascular cord integration whereas microtubule dissolution with nocodazole collapsed cord networks.These findings implicate VEGF-induction of calpain activity and impairment of cytoskeletal dynamics in the failure of VEGF-induced neovessels to form and

  1. Calpain Inhibition Reduces Axolemmal Leakage in Traumatic Axonal Injury

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    János Sándor

    2009-12-01

    Full Text Available Calcium-induced, calpain-mediated proteolysis (CMSP has recently been implicated to the pathogenesis of diffuse (traumatic axonal injury (TAI. Some studies suggested that subaxolemmal CMSP may contribute to axolemmal permeability (AP alterations observed in TAI. Seeking direct evidence for this premise we investigated whether subaxolemmal CMSP may contribute to axolemmal permeability alterations (APA and pre-injury calpain-inhibition could reduce AP in a rat model of TAI. Horseradish peroxidase (HRP, a tracer that accumulates in axons with APA was administered one hour prior to injury into the lateral ventricle; 30 min preinjury a single tail vein bolus injection of 30 mg/kg MDL-28170 (a calpain inhibitor or its vehicle was applied in Wistar rats exposed to impact acceleration brain injury. Histological detection of traumatically injured axonal segments accumulating HRP and statistical analysis revealed that pre-injury administration of the calpain inhibitor MDL-28170 significantly reduced the average length of HRP-labeled axonal segments. The axono-protective effect of pre-injury calpain inhibition recently demonstrated with classical immunohistochemical markers of TAI was further corroborated in this experiment; significant reduction of the length of labeled axons in the drug-treated rats implicate CMSP in the progression of altered AP in TAI.

  2. Vascular smooth muscle cell spreading onto fibrinogen is regulated by calpains and phospholipase C.

    Science.gov (United States)

    Paulhe, F; Bogyo, A; Chap, H; Perret, B; Racaud-Sultan, C

    2001-11-09

    Fibrinogen deposition and smooth muscle cell migration are important causes of atherosclerosis and angiogenesis. Involvement of calpains in vascular smooth muscle cell adhesion onto fibrinogen was investigated. Using calpain inhibitors, we showed that activation of calpains was required for smooth muscle cell spreading. An increase of (32)P-labeled phosphatidic acid and phosphatidylinositol-3,4-bisphosphate, respective products of phospholipase C and phosphoinositide 3-kinase activities, was measured in adherent cells. Addition of the calpain inhibitor calpeptin strongly decreased phosphatidic acid and phosphatidylinositol-3,4-bisphosphate. However, smooth muscle cell spreading was prevented by the phospholipase C inhibitor U-73122, but poorly modified by phosphoinositide 3-kinase inhibitors wortmannin and LY-294002. Moreover, PLC was found to act upstream of the PI 3-kinase IA isoform. Thus, our data provide the first evidence that calpains are required for smooth muscle cell spreading. Further, phospholipase C activation is pointed as a key step of cell-spreading regulation by calpains. Copyright 2001 Academic Press.

  3. Calpain Inhibition Improves Erectile Function in a Rat Model of Cavernous Nerve Injury.

    Science.gov (United States)

    Wan, Zhi-Hua; Li, Guo-Hao; Guo, Yong-Lian; Li, Wen-Zhou; Chen, Lin

    2015-01-01

    Erectile dysfunction (ED) after cavernous nerve (CN) injury remains difficult to treat. Calpain plays a critical role in causing neurodegenerative diseases. This study aimed to evaluate whether calpain inhibition preserves erectile function in a rat model of CN injury. Rats underwent sham surgery or CN crush injury. The CN-crushed rats were treated with vehicle or MDL-28170, a specific calpain inhibitor. At 1, 2, 3, and 7 days post-surgery, major pelvic ganglia (MPG) were harvested, followed by the measurement of erectile function, respectively. At 28 days, penile tissue and distal CN were harvested, followed by the measurement of erectile function in rats. Calpain activity in MPG and corpus cavernosum, as well as TGF-β1/Smad2 and collagen content in corpus cavernosum, were measured by western blot. Neuronal nitric oxide synthase (nNOS) was observed by immunohistochemistry. Increased calpain activity was observed in MPG and corpus cavernosum. CN crush markedly attenuated the erectile responses and nNOS expression in CN, and these were improved by MDL-28170 treatment. Furthermore, treatment prevented increased TGF-β1/Smad2 and collagen expression in corpus cavernosum. Our results suggested that calpain activation plays a role in pathogenesis of CN injury-associated ED. Calpain inhibition could be a novel approach for preventing the development of ED following CN injury. © 2015 S. Karger AG, Basel.

  4. Calpain 4 is not necessary for LFA-1-mediated function in CD4+ T cells.

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    Sarah A Wernimont

    2010-05-01

    Full Text Available T cell activation and immune synapse formation require the appropriate activation and clustering of the integrin, LFA-1. Previous work has reported that the calpain family of calcium-dependent proteases are important regulators of integrin activation and modulate T cell adhesion and migration. However, these studies have been limited by the use of calpain inhibitors, which have known off-target effects.Here, we used a LoxP/CRE system to specifically deplete calpain 4, a small regulatory calpain subunit required for expression and activity of ubiquitously expressed calpains 1 and 2, in CD4+ T cells. CD4+ and CD8+ T cells developed normally in Capn4(F/F:CD4-CRE mice and had severely diminished expression of Calpain 1 and 2, diminished talin proteolysis and impaired casein degradation. Calpain 4-deficient T cells showed no difference in adhesion or migration on the LFA-1 ligand ICAM-1 compared to control T cells. Moreover, there was no impairment in conjugation between Capn4(F/F:CD4-CRE T cells and antigen presenting cells, and the conjugates were still capable of polarizing LFA-1, PKC-theta and actin to the immune synapse. Furthermore, T cells from Capn4(F/F:CD4-CRE mice showed normal proliferation in response to either anti-CD3/CD28 coated beads or cognate antigen-loaded splenocytes. Finally, there were no differences in the rates of apoptosis following extrinsic and intrinsic apoptotic stimuli.Our findings demonstrate that calpain 4 is not necessary for LFA-1-mediated adhesion, conjugation or migration. These results challenge previous reports that implicate a central role for calpains in the regulation of T cell LFA-1 function.

  5. Calpain-like: A Ca(2+) dependent cystein protease in Entamoeba histolytica cell death.

    Science.gov (United States)

    Monroy, Virginia Sánchez; Flores, Olivia Medel; García, Consuelo Gómez; Maya, Yesenia Chávez; Fernández, Tania Domínguez; Pérez Ishiwara, D Guillermo

    2015-12-01

    Entamoeba histolytica programmed cell death (PCD) induced by G418 is characterized by the release of important amounts of intracellular calcium from reservoirs. Nevertheless, no typical caspases have been detected in the parasite, the PCD phenotype is inhibited by the cysteine protease inhibitor E-64. These results strongly suggest that Ca(2+)-dependent proteases could be involved in PCD. In this study, we evaluate the expression and activity of a specific dependent Ca(2+) protease, the calpain-like protease, by real-time quantitative PCR (RTq-PCR), Western blot assays and a enzymatic method during the induction of PCD by G418. Alternatively, using cell viability and TUNEL assays, we also demonstrated that the Z-Leu-Leu-Leu-al calpain inhibitor reduced the rate of cell death. The results demonstrated 4.9-fold overexpression of calpain-like gene 1.5 h after G418 PCD induction, while calpain-like protein increased almost two-fold with respect to basal calpain-like expression after 3 h of induction, and calpain activity was found to be approximately three-fold higher 6 h after treatment compared with untreated trophozoites. Taken together, these results suggest that this Ca(2+)-dependent protease could be involved in the executory phase of PCD. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Calpain-1 knockout reveals broad effects on erythrocyte deformability and physiology

    Science.gov (United States)

    Wieschhaus, Adam; Khan, Anwar; Zaidi, Asma; Rogalin, Henry; Hanada, Toshihiko; Liu, Fei; De Franceschi, Lucia; Brugnara, Carlo; Rivera, Alicia; Chishti, Athar H.

    2014-01-01

    Pharmacological inhibitors of cysteine proteases have provided useful insights into the regulation of calpain activity in erythrocytes. However, the precise biological function of calpain activity in erythrocytes remains poorly understood. Erythrocytes express calpain-1, an isoform regulated by calpastatin, the endogenous inhibitor of calpains. In the present study, we investigated the function of calpain-1 in mature erythrocytes using our calpain-1-null [KO (knockout)] mouse model. The calpain-1 gene deletion results in improved erythrocyte deformability without any measurable effect on erythrocyte lifespan in vivo. The calcium-induced sphero-echinocyte shape transition is compromised in the KO erythrocytes. Erythrocyte membrane proteins ankyrin, band 3, protein 4.1R, adducin and dematin are degraded in the calcium-loaded normal erythrocytes but not in the KO erythrocytes. In contrast, the integrity of spectrin and its state of phosphorylation are not affected in the calcium-loaded erythrocytes of either genotype. To assess the functional consequences of attenuated cytoskeletal remodelling in the KO erythrocytes, the activity of major membrane transporters was measured. The activity of the K+–Cl− co-transporter and the Gardos channel was significantly reduced in the KO erythrocytes. Similarly, the basal activity of the calcium pump was reduced in the absence of calmodulin in the KO erythrocyte membrane. Interestingly, the calmodulin-stimulated calcium pump activity was significantly elevated in the KO erythrocytes, implying a wider range of pump regulation by calcium and calmodulin. Taken together, and with the atomic force microscopy of the skeletal network, the results of the present study provide the first evidence for the physiological function of calpain-1 in erythrocytes with therapeutic implications for calcium imbalance pathologies such as sickle cell disease. PMID:22870887

  7. Calpain mediates pulmonary vascular remodeling in rodent models of pulmonary hypertension, and its inhibition attenuates pathologic features of disease

    Science.gov (United States)

    Ma, Wanli; Han, Weihong; Greer, Peter A.; Tuder, Rubin M.; Toque, Haroldo A.; Wang, Kevin K.W.; Caldwell, R. William; Su, Yunchao

    2011-01-01

    Pulmonary hypertension is a severe and progressive disease, a key feature of which is pulmonary vascular remodeling. Several growth factors, including EGF, PDGF, and TGF-β1, are involved in pulmonary vascular remodeling during pulmonary hypertension. However, increased knowledge of the downstream signaling cascades is needed if effective clinical interventions are to be developed. In this context, calpain provides an interesting candidate therapeutic target, since it is activated by EGF and PDGF and has been reported to activate TGF-β1. Thus, in this study, we examined the role of calpain in pulmonary vascular remodeling in two rodent models of pulmonary hypertension. These data showed that attenuated calpain activity in calpain-knockout mice or rats treated with a calpain inhibitor resulted in prevention of increased right ventricular systolic pressure, right ventricular hypertrophy, as well as collagen deposition and thickening of pulmonary arterioles in models of hypoxia- and monocrotaline-induced pulmonary hypertension. Additionally, inhibition of calpain in vitro blocked intracellular activation of TGF-β1, which led to attenuated Smad2/3 phosphorylation and collagen synthesis. Finally, smooth muscle cells of pulmonary arterioles from patients with pulmonary arterial hypertension showed higher levels of calpain activation and intracellular active TGF-β. Our data provide evidence that calpain mediates EGF- and PDGF-induced collagen synthesis and proliferation of pulmonary artery smooth muscle cells via an intracrine TGF-β1 pathway in pulmonary hypertension. PMID:22005303

  8. Calpain Inhibition Attenuates Adipose Tissue Inflammation and Fibrosis in Diet-induced Obese Mice.

    Science.gov (United States)

    Muniappan, Latha; Javidan, Aida; Jiang, Weihua; Mohammadmoradi, Shayan; Moorleghen, Jessica J; Katz, Wendy S; Balakrishnan, Anju; Howatt, Deborah A; Subramanian, Venkateswaran

    2017-10-31

    Adipose tissue macrophages have been proposed as a link between obesity and insulin resistance. However, the mechanisms underlying these processes are not completely defined. Calpains are calcium-dependent neutral cysteine proteases that modulate cellular function and have been implicated in various inflammatory diseases. To define whether activated calpains influence diet-induced obesity and adipose tissue macrophage accumulation, mice that were either wild type (WT) or overexpressing calpastatin (CAST Tg), the endogenous inhibitor of calpains were fed with high (60% kcal) fat diet for 16 weeks. CAST overexpression did not influence high fat diet-induced body weight and fat mass gain throughout the study. Calpain inhibition showed a transient improvement in glucose tolerance at 5 weeks of HFD whereas it lost this effect on glucose and insulin tolerance at 16 weeks HFD in obese mice. However, CAST overexpression significantly reduced adipocyte apoptosis, adipose tissue collagen and macrophage accumulation as detected by TUNEL, Picro Sirius and F4/80 immunostaining, respectively. CAST overexpression significantly attenuated obesity-induced inflammatory responses in adipose tissue. Furthermore, calpain inhibition suppressed macrophage migration to adipose tissue in vitro. The present study demonstrates a pivotal role for calpains in mediating HFD-induced adipose tissue remodeling by influencing multiple functions including apoptosis, fibrosis and inflammation.

  9. Disruption of calpain reduces lipotoxicity-induced cardiac injury by preventing endoplasmic reticulum stress.

    Science.gov (United States)

    Li, Shengcun; Zhang, Lulu; Ni, Rui; Cao, Ting; Zheng, Dong; Xiong, Sidong; Greer, Peter A; Fan, Guo-Chang; Peng, Tianqing

    2016-11-01

    Diabetes and obesity are prevalent in westernized countries. In both conditions, excessive fatty acid uptake by cardiomyocytes induces cardiac lipotoxicity, an important mechanism contributing to diabetic cardiomyopathy. This study investigated the effect of calpain disruption on cardiac lipotoxicity. Cardiac-specific capns1 knockout mice and their wild-type littermates (male, age of 4weeks) were fed a high fat diet (HFD) or normal diet for 20weeks. HFD increased body weight, altered blood lipid profiles and impaired glucose tolerance comparably in both capns1 knockout mice and their wild-type littermates. Calpain activity, cardiomyocyte cross-sectional areas, collagen deposition and triglyceride were significantly increased in HFD-fed mouse hearts, and these were accompanied by myocardial dysfunction and up-regulation of hypertrophic and fibrotic collagen genes as well as pro-inflammatory cytokines. These effects of HFD were attenuated by disruption of calpain in capns1 knockout mice. Mechanistically, deletion of capns1 in HFD-fed mouse hearts and disruption of calpain with calpain inhibitor-III, silencing of capn1, or deletion of capns1 in palmitate-stimulated cardiomyocytes prevented endoplasmic reticulum stress, apoptosis, cleavage of caspase-12 and junctophilin-2, and pro-inflammatory cytokine expression. Pharmacological inhibition of endoplasmic reticulum stress diminished palmitate-induced apoptosis and pro-inflammatory cytokine expression in cardiomyocytes. In summary, disruption of calpain prevents lipotoxicity-induced apoptosis in cardiomyocytes and cardiac injury in mice fed a HFD. The role of calpain is mediated, at least partially, through endoplasmic reticulum stress. Thus, calpain/endoplasmic reticulum stress may represent a new mechanism and potential therapeutic targets for cardiac lipotoxicity. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Accumulation of human full-length tau induces degradation of nicotinic acetylcholine receptor α4 via activating calpain-2.

    Science.gov (United States)

    Yin, Yaling; Wang, Yali; Gao, Di; Ye, Jinwang; Wang, Xin; Fang, Lin; Wu, Dongqin; Pi, Guilin; Lu, Chengbiao; Zhou, Xin-Wen; Yang, Ying; Wang, Jian-Zhi

    2016-06-09

    Cholinergic impairments and tau accumulation are hallmark pathologies in sporadic Alzheimer's disease (AD), however, the intrinsic link between tau accumulation and cholinergic deficits is missing. Here, we found that overexpression of human wild-type full-length tau (termed hTau) induced a significant reduction of α4 subunit of nicotinic acetylcholine receptors (nAChRs) with an increased cleavage of the receptor producing a ~55kDa fragment in primary hippocampal neurons and in the rat brains, meanwhile, the α4 nAChR currents decreased. Further studies demonstrated that calpains, including calpain-1 and calpain-2, were remarkably activated with no change of caspase-3, while simultaneous suppression of calpain-2 by selective calpain-2 inhibitor but not calpain-1 attenuated the hTau-induced degradation of α4 nAChR. Finally, we demonstrated that hTau accumulation increased the basal intracellular calcium level in primary hippocampal neurons. We conclude that the hTau accumulation inhibits nAChRs α4 by activating calpain-2. To our best knowledge, this is the first evidence showing that the intracellular accumulation of tau causes cholinergic impairments.

  11. Increased μ-Calpain Activity in Blasts of Common B-Precursor Childhood Acute Lymphoblastic Leukemia Correlates with Their Lower Susceptibility to Apoptosis.

    Directory of Open Access Journals (Sweden)

    Anna Mikosik

    Full Text Available Childhood acute lymphoblastic leukemia (ALL blasts are characterized by inhibited apoptosis promoting fast disease progress. It is known that in chronic lymphocytic and acute myeloid leukemias the reduced apoptosis is strongly related with the activity of calpain-calpastatin system (CCS composed of cytoplasmic proteases--calpains--performing the modulatory proteolysis of key proteins involved in cell proliferation and apoptosis, and of their endogenous inhibitor--calpastatin. Here, the CCS protein abundance and activity was for the first time studied in childhood ALL blasts and in control bone marrow CD19+ B cells by semi-quantitative flow cytometry and western blotting of calpastatin fragments resulting from endogenous calpain activity. Significantly higher μ-calpain (CAPN1 gene transcription, protein amounts and activity (but not those of m-calpain, with calpastatin amount and transcription of its gene (CAST greatly varying were observed in CD19(+ ALL blasts compared to control cells. Significant inverse relation between the amount/activity of calpain and spontaneous apoptosis was noted. Patients older than 10 years (considered at higher risk displayed increased amounts and activities of blast calpain. Finally, treatment of blasts with the tripeptide calpain inhibitors II and IV significantly and in dose-dependent fashion increased the percentage of blasts entering apoptosis. Together, these findings make the CCS a potential new predictive tool and therapeutic target in childhood ALL.

  12. Calpain 3 is important for muscle regeneration

    DEFF Research Database (Denmark)

    Hauerslev, Simon; Sveen, Marie-Louise; Duno, Morten

    2012-01-01

    Limb girdle muscular dystrophy (LGMD) type 2A is caused by mutations in the CAPN3 gene and complete lack of functional calpain 3 leads to the most severe muscle wasting. Calpain 3 is suggested to be involved in maturation of contractile elements after muscle degeneration. The aim of this study...

  13. The Sirtuin 2 Inhibitor AK-7 Is Neuroprotective in Huntington’s Disease Mouse Models

    Directory of Open Access Journals (Sweden)

    Vanita Chopra

    2012-12-01

    Full Text Available Inhibition of sirtuin 2 (SIRT2 deacetylase mediates protective effects in cell and invertebrate models of Parkinson’s disease and Huntington’s disease (HD. Here we report the in vivo efficacy of a brain-permeable SIRT2 inhibitor in two genetic mouse models of HD. Compound treatment resulted in improved motor function, extended survival, and reduced brain atrophy and is associated with marked reduction of aggregated mutant huntingtin, a hallmark of HD pathology. Our results provide preclinical validation of SIRT2 inhibition as a potential therapeutic target for HD and support the further development of SIRT2 inhibitors for testing in humans.

  14. Evidence supporting the role of calpain in the α-processing of amyloid-β precursor protein.

    Science.gov (United States)

    Nguyen, Huey T; Sawmiller, Darrell R; Wu, Qi; Maleski, Jerome J; Chen, Ming

    2012-04-13

    Amyloid plaques are a hallmark of the aging and senile dementia brains, yet their mechanism of origins has remained elusive. A central issue is the regulatory mechanism and identity of α-secretase, a protease responsible for α-processing of amyloid-β precursor protein (APP). A remarkable feature of this enzyme is its high sensitivity to a wide range of cellular stimulators, many of which are agonists for Ca(2+) signaling. This feature, together with previous work in our laboratory, has suggested that calpain, a Ca(2+)-dependent protease, plays a key role in APP α-processing. In this study we report that overexpression of the μ-calpain gene in HEK293 cells resulted in a 2.7-fold increase of the protein levels. Measurements of intracellular calpain enzymatic activity revealed that the calpain overexpressing cells displayed a prominent elevation of the activity compared to wild-type cells. When the cells were stimulated by nicotine, glutamate or phorbol 12,13-dibutylester, the activity increase was even more remarkable and sensitive to calpeptin, a calpain inhibitor. Meanwhile, APP secretion from the calpain overexpressing cells was robustly increased under both resting and stimulated conditions over wild-type cells. Furthermore, cell surface biotinylation experiments showed that μ-calpain was clearly detected among the cell surface proteins. These data together support our view that calpain should be a reasonable candidate for α-secretase for further study. This model is discussed with an interesting fact that three other deposited proteins (tau, spectrin and crystalline) are also the known substrates of calpain. Finally we discuss some current misconceptions in senile dementia research. Published by Elsevier Inc.

  15. A possible therapeutic potential of quercetin through inhibition of μ-calpain in hypoxia induced neuronal injury: a molecular dynamics simulation study

    Directory of Open Access Journals (Sweden)

    Anand Kumar Pandey

    2016-01-01

    Full Text Available The neuroprotective property of quercetin is well reported against hypoxia and ischemia in past studies. This property of quercetin lies in its antioxidant property with blood-brain barrier permeability and anti-inflammatory capabilities. µ-Calpain, a calcium ion activated intracellular cysteine protease causes serious cellular insult, leading to cell death in various pathological conditions including hypoxia and ischemic stroke. Hence, it may be considered as a potential drug target for the treatment of hypoxia induced neuronal injury. As the inhibitory property of µ-calpain is yet to be explored in details, hence, in the present study, we investigated the interaction of quercetin with µ-calpain through a molecular dynamics simulation study as a tool through clarifying the molecular mechanism of such inhibition and determining the probable sites and modes of quercetin interaction with the µ-calpain catalytic domain. In addition, we also investigated the structure-activity relationship of quercetin with μ-calpain. Affinity binding of quercetin with µ-calpain had a value of –28.73 kJ/mol and a Ki value of 35.87 µM that may be a probable reason to lead to altered functioning of µ-calpain. Hence, quercetin was found to be an inhibitor of µ-calpain which might have a possible therapeutic role in hypoxic injury.

  16. Calpain-mediated cleavage of DARPP-32 in Alzheimer's disease.

    Science.gov (United States)

    Cho, Kwangmin; Cho, Mi-Hyang; Seo, Jung-Han; Peak, Jongjin; Kong, Kyoung-Hye; Yoon, Seung-Yong; Kim, Dong-Hou

    2015-10-01

    Toxicity induced by aberrant protein aggregates in Alzheimer's disease (AD) causes synaptic disconnection and concomitant progressive neurodegeneration that eventually impair cognitive function. cAMP-response element-binding protein (CREB) is a transcription factor involved in the molecular switch that converts short-term to long-term memory. Although disturbances in CREB function have been suggested to cause memory deficits in both AD and AD animal models, the mechanism of CREB dysfunction is still unclear. Here, we show that the dopamine- and cAMP-regulated phosphoprotein 32 kDa (DARPP-32), a key inhibitor of protein phosphate-1 (PP-1) that regulates CREB phosphorylation, is cleaved by activated calpain in both AD brains and neuronal cells treated with amyloid-β or okadaic acid, a protein phosphatase-2A inhibitor that induces tau hyperphosphorylation and neuronal death. We found that DARPP-32 is mainly cleaved at Thr(153) by calpain and that this cleavage of DARPP-32 reduces CREB phosphorylation via loss of its inhibitory function on PP1. Our results suggest a novel mechanism of DARPP-32-CREB signalling dysregulation in AD. © 2015 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.

  17. Bcr-abl regulates Stat5 through Shp2, the interferon consensus sequence binding protein (Icsbp/Irf8), growth arrest specific 2 (Gas2) and calpain

    Science.gov (United States)

    Hjort, Elizabeth E.; Huang, Weiqi; Hu, Liping; Eklund, Elizabeth A.

    2016-01-01

    Icsbp/Irf8 is an interferon regulatory transcription factor that functions as a suppressor of myeloid leukemias. Consistent with this activity, Icsbp represses a set of genes encoding proteins that promote cell proliferation/survival. One such gene encodes Gas2, a calpain inhibitor. We previously found that increased Gas2-expression in Bcr-abl+ cells stabilized βcatenin; a Calpain substrate. This was of interest, because βcatenin contributes to disease progression in chronic myeloid leukemia (CML). Calpain has additional substrates implicated in leukemogenesis, including Stat5. In the current study, we hypothesized that Stat5 activity in CML is regulated by Gas2/Calpain. We found that Bcr-abl-induced, Shp2-dependent dephosphorylation of Icsbp impaired repression of GAS2 by this transcription factor. The consequent decrease in Calpain activity stabilized Stat5 protein; increasing the absolute abundance of both phospho and total Stat5. This enhanced repression of the IRF8 promoter by Stat5 in a manner dependent on Icsbp, Gas2 and Calpain, but not Stat5 tyrosine phosphorylation. During normal myelopoiesis, increased expression and phosphorylation of Icsbp inhibits Calpain. In contrast, constitutive activation of Shp2 in Bcr-abl+ cells impairs regulation of Gas2/Calpain by Icsbp, aberrantly stabilizing Stat5 and enhancing IRF8 repression. This novel feedback mechanism enhances leukemogenesis by increasing Stat5 and decreasing Icsbp. Bcr-abl targeted tyrosine kinase inhibitors (TKIs) provide long term disease control, but CML is not cured by these agents. Our studies suggest targeting Calpain might be a rational therapeutic approach to decrease persistent leukemia stem cells (LSCs) during TKI-treatment. PMID:27769062

  18. Dual Vulnerability of Tau to Calpains and Caspase-3 Proteolysis Under Neurotoxic and Neurodegenerative Conditions

    Directory of Open Access Journals (Sweden)

    Ming Cheng Liu

    2010-11-01

    Full Text Available Axonally specific microtubule-associated protein tau is an important component of neurofibrillary tangles found in AD (Alzheimer's disease and other tauopathy diseases such as CTE (chronic traumatic encephalopathy. Such tau aggregate is found to be hyperphosphorylated and often proteolytically fragmented. Similarly, tau is degraded following TBI (traumatic brain injury. In the present study, we examined the dual vulnerability of tau to calpain and caspase-3 under neurotoxic and neurodegenerative conditions. We first identified three novel calpain cleavage sites in rat tau (four-repeat isoform as Ser130 ↓ Lys131, Gly157 ↓ Ala158 and Arg380 ↓ Glu381. Fragment-specific antibodies to target the major calpain-mediated TauBDP-35K (35 kDa tau-breakdown product and the caspase-mediated TauBDP-45K respectively were developed. In rat cerebrocortical cultures treated with excitotoxin [NMDA (N-methyl-D-aspartate], tau is significantly degraded into multiple fragments, including a dominant signal of calpain-mediated TauBDP-35K with minimal caspase-mediated TauBDP-45K. Following apoptosis-inducing EDTA treatment, tau was truncated only to TauBDP-48K/45K-exclusively by caspase. Cultures treated with another apoptosis inducer STS (staurosporine, dual fragmentation by calpain (TauBDP-35K and caspase-3 (TauBDP-45K was observed. Tau was also fragmented in injured rat cortex following TBI in vivo to BDPs of 45-42 kDa (minor, 35 kDa and 15 kDa, followed by TauBDP-25K. Calpain-mediated TauBDP-35K-specific antibody confirmed robust signals in the injured cortex, while caspase-mediated TauBDP-45K-specific antibody only detected faint signals. Furthermore, intravenous administration of a calpain-specific inhibitor SNJ-1945 strongly suppressed the TauBDP-35K formation. Taken together, these results suggest that tau protein is dually vulnerable to calpain and caspase-3 proteolysis under different neurotoxic and injury conditions.

  19. Activation of calpain-1 in human carotid artery atherosclerotic lesions

    Directory of Open Access Journals (Sweden)

    Pedro Luis M

    2009-06-01

    Full Text Available Abstract Background In a previous study, we observed that oxidized low-density lipoprotein-induced death of endothelial cells was calpain-1-dependent. The purpose of the present paper was to study the possible activation of calpain in human carotid plaques, and to compare calpain activity in the plaques from symptomatic patients with those obtained from patients without symptoms. Methods Human atherosclerotic carotid plaques (n = 29, 12 associated with symptoms were removed by endarterectomy. Calpain activity and apoptosis were detected by performing immunohistochemical analysis and TUNEL assay on human carotid plaque sections. An antibody specific for calpain-proteolyzed α-fodrin was used on western blots. Results We found that calpain was activated in all the plaques and calpain activity colocalized with apoptotic cell death. Our observation of autoproteolytic cleavage of the 80 kDa subunit of calpain-1 provided further evidence for enzyme activity in the plaque samples. When calpain activity was quantified, we found that plaques from symptomatic patients displayed significantly lower calpain activity compared with asymptomatic plaques. Conclusion These novel results suggest that calpain-1 is commonly active in carotid artery atherosclerotic plaques, and that calpain activity is colocalized with cell death and inversely associated with symptoms.

  20. The sirtuin-2 inhibitor AK7 is neuroprotective in models of Parkinson's disease but not amyotrophic lateral sclerosis and cerebral ischemia.

    Directory of Open Access Journals (Sweden)

    Xiqun Chen

    Full Text Available Sirtuin deacetylases regulate diverse cellular pathways and influence disease processes. Our previous studies identified the brain-enriched sirtuin-2 (SIRT2 deacetylase as a potential drug target to counteract neurodegeneration. In the present study, we characterize SIRT2 inhibition activity of the brain-permeable compound AK7 and examine the efficacy of this small molecule in models of Parkinson's disease, amyotrophic lateral sclerosis and cerebral ischemia. Our results demonstrate that AK7 is neuroprotective in models of Parkinson's disease; it ameliorates alpha-synuclein toxicity in vitro and prevents 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP-induced dopamine depletion and dopaminergic neuron loss in vivo. The compound does not show beneficial effects in mouse models of amyotrophic lateral sclerosis and cerebral ischemia. These findings underscore the specificity of protective effects observed here in models of Parkinson's disease, and previously in Huntington's disease, and support the development of SIRT2 inhibitors as potential therapeutics for the two neurodegenerative diseases.

  1. Phosphorylation prevents C/EBP{beta} from the calpain-dependent degradation

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Yuan-yuan; Li, Shu-fen; Qian, Shu-wen; Zhang, You-you; Liu, Yuan; Tang, Qi-Qun; Li, Xi, E-mail: lixi@shmu.edu.cn

    2012-03-16

    Highlights: Black-Right-Pointing-Pointer Phosphorylation protected C/EBP{beta} from {mu}-calpain-mediated proteolysis in vitro. Black-Right-Pointing-Pointer Phosphorylation mimic C/EBP{beta} was insensitive to calpain accelerator and inhibitor. Black-Right-Pointing-Pointer Phosphorylation on Thr{sub 188} contributed more to the stabilization of C/EBP{beta}. -- Abstract: CCAAT/enhancer-binding protein (C/EBP) {beta} plays an important role in proliferation and differentiation of 3T3-L1 preadipocytes. C/EBP{beta} is sequentially phosphorylated during the 3T3-L1 adipocyte differentiation program, first by MAPK/Cyclin A/cdk2 on Thr{sub 188} and subsequently by GSK3{beta} on Ser{sub 184} or Thr{sub 179}. Dual phosphorylation is critical for the gain of DNA binding activity of C/EBP{beta}. In this manuscript, we found that phosphorylation also contributed to the stability of C/EBP{beta}. Both ex vivo and in vitro experiments showed that phosphorylation by MAPK/Cyclin A/cdk2 and GSK3{beta} protected C/EBP{beta} from {mu}-calpain-mediated proteolysis, while phosphorylation on Thr{sub 188} by MAPK/Cyclin A/cdk2 contributed more to the stabilization of C/EBP{beta}, Further studies indicated that phosphorylation mimic C/EBP{beta} was insensitive to both calpain accelerator and calpain inhibitor. Thus, phosphorylation might contribute to the stability as well as the gain of DNA binding activity of C/EBP{beta}.

  2. Extracellular Calpain/Calpastatin Balance Is Involved in the Progression of Pulmonary Hypertension.

    Science.gov (United States)

    Wan, Feng; Letavernier, Emmanuel; Abid, Shariq; Houssaini, Amal; Czibik, Gabor; Marcos, Elisabeth; Rideau, Dominique; Parpaleix, Aurélien; Lipskaia, Larissa; Amsellem, Valérie; Gellen, Barnabas; Sawaki, Daigo; Derumeaux, Genevieve; Dubois-Randé, Jean-Luc; Delcroix, Marion; Quarck, Rozenn; Baud, Laurent; Adnot, Serge

    2016-09-01

    Excessive growth of pulmonary arterial (PA) smooth muscle cells (SMCs) is a major component of PA hypertension (PAH). The calcium-activated neutral cysteine proteases calpains 1 and 2, expressed by PASMCs, contribute to PH but are tightly controlled by a single specific inhibitor, calpastatin. Our objective was to investigate calpastatin during pulmonary hypertension (PH) progression and its potential role as an intracellular and/or extracellular effector. We assessed calpains and calpastatin in patients with idiopathic PAH and mice with hypoxic or spontaneous (SM22-5HTT(+) strain) PH. To assess intracellular and extracellular roles for calpastatin, we studied effects of the calpain inhibitor PD150606 on hypoxic PH in mice with calpastatin overexpression driven by the cytomegalovirus promoter (CMV-Cast) or C-reactive protein (CRP) promoter (CRP-Cast), inducing increased calpastatin production ubiquitously and in the liver, respectively. Chronically hypoxic and SM22-5HTT(+) mice exhibited increased lung calpastatin and calpain 1 and 2 protein levels and activity, both intracellularly and extracellularly. Prominent calpastatin and calpain immunostaining was found in PASMCs of remodeled vessels in mice and patients with PAH, who also exhibited increased plasma calpastatin levels. CMV-Cast and CRP-Cast mice showed similarly decreased PH severity compared with wild-type mice, with no additional effect of PD150606 treatment. In cultured PASMCs from wild-type and CMV-Cast mice, exogenous calpastatin decreased cell proliferation and migration with similar potency as PD150606 and suppressed fibronectin-induced potentiation. These results indicate that calpastatin limits PH severity via extracellular mechanisms. They suggest a new approach to the development of treatments for PH.

  3. Conditional disruption of calpain in the CNS alters dendrite morphology, impairs LTP, and promotes neuronal survival following injury

    National Research Council Canada - National Science Library

    Amini, Mandana; Ma, Chun-lei; Farazifard, Rasoul; Zhu, Guoqi; Zhang, Yi; Vanderluit, Jacqueline; Zoltewicz, Joanna Susie; Hage, Fadi; Savitt, Joseph M; Lagace, Diane C; Slack, Ruth S; Beique, Jean-Claude; Baudry, Michel; Greer, Peter A; Bergeron, Richard; Park, David S

    2013-01-01

    Ubiquitous classical (typical) calpains, calpain-1 and calpain-2, are Ca(+2)-dependent cysteine proteases, which have been associated with numerous physiological and pathological cellular functions...

  4. Lesions of entorhinal cortex produce a calpain-mediated degradation of brain spectrin in dentate gyrus. I. Biochemical studies.

    Science.gov (United States)

    Seubert, P; Ivy, G; Larson, J; Lee, J; Shahi, K; Baudry, M; Lynch, G

    1988-09-06

    Lesions of the rat entorhinal cortex cause extensive synaptic restructuring and perturbation of calcium regulation in the dentate gyrus of hippocampus. Calpain is a calcium-activated protease which has been implicated in degenerative phenomena in muscles and in peripheral nerves. In addition, calpain degrades several major structural neuronal proteins and has been proposed to play a critical role in the morphological changes observed following deafferentation. In this report we present evidence that lesions of the entorhinal cortex produce a marked increase in the breakdown of brain spectrin, a substrate for calpain, in the dentate gyrus. Two lines of evidence indicate that this effect is due to calpain activation: (i) the spectrin breakdown products observed following the lesion are indistinguishable from calpain-generated spectrin fragments in vitro; and (ii) their appearance can be reduced by prior intraventricular in fusion of leupeptin, a calpain inhibitor. Levels of spectrin breakdown products are increased as early as 4 h post-lesion, reach maximal values at 2 days, and remain above normal to some degree for at least 27 days. In addition, a small but significant increase in spectrin proteolysis is also observed in the hippocampus contralateral to the lesioned side in the first week postlesion. At 2 days postlesion the total spectrin immunoreactivity (native polypeptide plus breakdown products) increases by 40%, suggesting that denervation of the dentate gyrus produces not only an increased rate of spectrin degradation but also an increased rate of spectrin synthesis. These results indicate that calpain activation and spectrin degradation are early biochemical events following deafferentation and might well participate in the remodelling of postsynaptic structures. Finally, the magnitude of the observed effects as well as the stable nature of the breakdown products provide a sensitive assay for neuronal pathology.

  5. Acetylcholine Attenuated TNF-α-Induced Apoptosis in H9c2 Cells: Role of Calpain and the p38-MAPK Pathway

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    Ming Zhao

    2015-07-01

    Full Text Available Background: Previous studies have shown that inflammation is associated with excessive activation of calpains. Acetylcholine (ACh has been reported to inhibit pro-inflammatory cytokine release and protect against cardiomyocyte injury. However, there is no direct evidence regarding whether ACh can regulate calpains to exert cardioprotection. To this end, we investigated the effect of ACh on tumour necrosis factor alpha (TNF-α-induced cardiomyocyte injury and further explored the underlying mechanism. Methods: Flow cytometry and transmission electron microscopy were performed to evaluate apoptosis and cellular ultrastructure. Western blotting was performed to assess changes in protein expression. siRNA was employed to silence specific proteins. Results: TNF-α treatment increased the expression of cleaved caspase-3, calpain-1 and p38-mitogen-activated protein kinase (p38-MAPK. The calpain inhibitor PD150606 and the p38-MAPK inhibitor SB203580 inhibited apoptosis induced by TNF-α. Moreover, SB203580 decreased the expression and activity of calpain-1, possibly related to the up-regulation of calpastatin. ACh significantly inhibited TNF-α-induced cell apoptosis, as evidenced by decreases in caspase-3 cleavage, p38-MAPK phosphorylation, and calpain-1 expression and activity as well as increases in calpastatin expression. These beneficial effects of ACh were abolished by atropine or M2AChR siRNA. Conclusion: Our results suggest that ACh ameliorated TNF-α-induced calpain activation by decreasing p38-MAPK phosphorylation and enhancing calpastatin expression, indicating that calpain may be an important link between inflammatory factors and myocardial cell apoptosis.

  6. Inhibitors

    Science.gov (United States)

    ... and exercise, immune tolerance therapy, and needs of older adults with hemophilia and an inhibitor. For more information, visit https://www.hemophilia.org/Events-Educational-Programs/Inhibitor-Education/Inhibitor-Education-Summits The NHF’s Inhibitor Education Summits ...

  7. Psiguajadials A-K: Unusual Psidium Meroterpenoids as Phosphodiesterase-4 Inhibitors from the Leaves of Psidium guajava.

    Science.gov (United States)

    Tang, Gui-Hua; Dong, Zhen; Guo, Yan-Qiong; Cheng, Zhong-Bin; Zhou, Chu-Jun; Yin, Sheng

    2017-04-21

    Bioassay-guided fractionation of the ethanolic extract of the leaves of Psidium guajava led to the isolation of 11 new Psidium meroterpenoids, psiguajadials A-K (1-11), along with 17 known ones (12-28). Their structures and absolute configurations were elucidated by spectroscopic methods and comparison of experimental and calculated ECD. Compounds 1 and 2 represent two unprecedented skeletons of 3,5-diformyl-benzyl phloroglucinol-coupled sesquiterpenoid, while 3 is the first example of Psidium meroterpenoids coupling via an oxepane ring. Putative biosynthetic pathways towards 1 and 2 are proposed. Compounds 1-13 and 16-26 exhibited moderate inhibitory activities against phosphodiesterase-4 (PDE4), a drug target for asthma and chronic obstructive pulmonary disease, with IC 50 values in the range of 1.34-7.26 μM.

  8. Calpain 3 is a rapid-action, unidirectional proteolytic switch central to muscle remodeling.

    Science.gov (United States)

    de Morrée, Antoine; Lutje Hulsik, David; Impagliazzo, Antonietta; van Haagen, Herman H H B M; de Galan, Paula; van Remoortere, Alexandra; 't Hoen, Peter A C; van Ommen, Gertjan B; Frants, Rune R; van der Maarel, Silvère M

    2010-08-04

    Calpain 3 (CAPN3) is a cysteine protease that when mutated causes Limb Girdle Muscular Dystrophy 2A. It is thereby the only described Calpain family member that genetically causes a disease. Due to its inherent instability little is known of its substrates or its mechanism of activity and pathogenicity. In this investigation we define a primary sequence motif underlying CAPN3 substrate cleavage. This motif can transform non-related proteins into substrates, and identifies >300 new putative CAPN3 targets. Bioinformatic analyses of these targets demonstrate a critical role in muscle cytoskeletal remodeling and identify novel CAPN3 functions. Among the new CAPN3 substrates are three E3 SUMO ligases of the Protein Inhibitor of Activated Stats (PIAS) family. CAPN3 can cleave PIAS proteins and negatively regulates PIAS3 sumoylase activity. Consequently, SUMO2 is deregulated in patient muscle tissue. Our study thus uncovers unexpected crosstalk between CAPN3 proteolysis and protein sumoylation, with strong implications for muscle remodeling.

  9. Simvastatin inhibited oxLDL-induced proatherogenic effects through calpain-1-PPARγ-CD36 pathway.

    Science.gov (United States)

    Yang, Xueyan; Yin, Meihui; Yu, Lan; Lu, Meili; Wang, Hongxin; Tang, Futian; Zhang, Yingjie

    2016-12-01

    We previously reported that simvastatin, an inhibitor of HMG-CoA reductase, inhibits atherosclerosis in rats. The present study was designed to investigate the effect of simvastatin on mouse peritoneal macrophage foam cell formation, the early feature of atherosclerosis, and explore its mechanisms. The results showed that simvastatin decreased cholesterol content and DiI-oxLDL (1,1'-didodecyl 3,3,3',3'-indocarbocyanine perchlorate - oxidized low-density lipoprotein) uptake, reduced the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the medium, down-regulated the mRNA and protein expression of CD36 (a fatty acid receptor), and reduced the mRNA expressions of peroxisome proliferator-activated receptor gamma (PPARγ), TNF-α, and IL-6 in macrophages treated with oxLDL. However, PPARγ agonist troglitazone partly abolished the effects of simvastatin on foam cells. In addition, simvastatin reduced the protein expression of calpain-1, a Ca(2+)-sensitive cysteine protease, in oxLDL-treated macrophages. Furthermore, PD150606, a specific calpain inhibitor, reduced mRNA expressions of PPARγ and CD36 in macrophages treated with oxLDL. Combination of simvastatin and PD150606 had no further effect on mRNA expression of PPARγ and CD36 compared with either alone. However, over-expression of calpain-1 in macrophages partly reversed the simvastatin effects, including cell cholesterol content, mRNA expressions of PPARγ, and CD36. The results suggested that simvastatin inhibits foam cell formation of oxLDL-treated macrophages through a calpain-1-PPARγ-CD36 pathway.

  10. Bid and calpains cooperate to trigger oxaliplatin-induced apoptosis of cervical carcinoma HeLa cells.

    Science.gov (United States)

    Anguissola, Sergio; Köhler, Barbara; O'Byrne, Robert; Düssmann, Heiko; Cannon, Mary D; Murray, Frank E; Concannon, Caoimhin G; Rehm, Markus; Kögel, Donat; Prehn, Jochen H M

    2009-11-01

    The Bcl-2 homology 3-only protein Bid is an important mediator of death receptor-induced apoptosis. Recent reports and this study suggest that Bid may also mediate genotoxic drug-induced apoptosis of various human cancer cells. Here, we characterized the role of Bid and the mechanism of Bid activation during oxaliplatin-induced apoptosis of HeLa cervical cancer cells. Small hairpin RNA-mediated silencing of Bid protected HeLa cells against both death receptor- and oxaliplatin-induced apoptosis. Expression of a Bid mutant in which caspase-8 cleavage site was mutated (D59A) reactivated oxaliplatin-induced apoptosis in Bid-deficient cells but failed to reactivate death receptor-induced apoptosis, suggesting that caspase-8-mediated Bid cleavage did not contribute to oxaliplatin-induced apoptosis. Overexpression of bcl-2 or treatment with the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-dl-Asp-fluoromethylketone abolished caspase-2, -8, -9, and -3 activation as well as Bid cleavage in response to oxaliplatin, suggesting that Bid cleavage occurred downstream of mitochondrial permeabilization and was predominantly mediated by caspases. We also detected an early activation of calpains in response to oxaliplatin. Calpain inhibition reduced Bid cleavage, mitochondrial depolarization, and activation of caspase-9, -3, -2, and -8 in response to oxaliplatin. Further experiments, however, suggested that Bid cleavage by calpains was not a prerequisite for oxaliplatin-induced apoptosis: single-cell imaging experiments using a yellow fluorescent protein-Bid-cyan fluorescent protein probe demonstrated translocation of full-length Bid to mitochondria that was insensitive to calpain or caspase inhibition. Moreover, calpain inhibition showed a potent protective effect in Bid-silenced cells. In conclusion, our data suggest that calpains and Bid act in a cooperative, but mutually independent, manner to mediate oxaliplatin-induced apoptosis of HeLa cells.

  11. Massive expansion of the calpain gene family in unicellular eukaryotes

    Directory of Open Access Journals (Sweden)

    Zhao Sen

    2012-09-01

    Full Text Available Abstract Background Calpains are Ca2+-dependent cysteine proteases that participate in a range of crucial cellular processes. Dysfunction of these enzymes may cause, for instance, life-threatening diseases in humans, the loss of sex determination in nematodes and embryo lethality in plants. Although the calpain family is well characterized in animal and plant model organisms, there is a great lack of knowledge about these genes in unicellular eukaryote species (i.e. protists. Here, we study the distribution and evolution of calpain genes in a wide range of eukaryote genomes from major branches in the tree of life. Results Our investigations reveal 24 types of protein domains that are combined with the calpain-specific catalytic domain CysPc. In total we identify 41 different calpain domain architectures, 28 of these domain combinations have not been previously described. Based on our phylogenetic inferences, we propose that at least four calpain variants were established in the early evolution of eukaryotes, most likely before the radiation of all the major supergroups of eukaryotes. Many domains associated with eukaryotic calpain genes can be found among eubacteria or archaebacteria but never in combination with the CysPc domain. Conclusions The analyses presented here show that ancient modules present in prokaryotes, and a few de novo eukaryote domains, have been assembled into many novel domain combinations along the evolutionary history of eukaryotes. Some of the new calpain genes show a narrow distribution in a few branches in the tree of life, likely representing lineage-specific innovations. Hence, the functionally important classical calpain genes found among humans and vertebrates make up only a tiny fraction of the calpain family. In fact, a massive expansion of the calpain family occurred by domain shuffling among unicellular eukaryotes and contributed to a wealth of functionally different genes.

  12. Calpain-2 Regulates TNF-α Expression Associated with Neuropathic Pain Following Motor Nerve Injury.

    Science.gov (United States)

    Chen, Shao-Xia; Liao, Guang-Jie; Yao, Pei-Wen; Wang, Shao-Kun; Li, Yong-Yong; Zeng, Wei-An; Liu, Xian-Guo; Zang, Ying

    2018-02-22

    Both calpain-2 (CALP2) and tumor necrosis factor-α (TNF-α) contribute to persistent bilateral hypersensitivity in animals subjected to L5 ventral root transection (L5-VRT), a model of selective motor fiber injury without sensory nerve damage. However, specific upstream mechanisms regulating TNF-α overexpression and possible relationships linking CALP2 and TNF-α have not yet been investigated in this model. We examined changes in CALP2 and TNF-α protein levels and alterations in bilateral mechanical threshold within 24 h following L5-VRT model injury. We observed robust elevation of CALP2 and TNF-α in bilateral dorsal root ganglias (DRGs) and bilateral spinal cord neurons. CALP2 and TNF-α protein induction by L5-VRT were significantly inhibited by pretreatment using the calpain inhibitor MDL28170. Administration of CALP2 to rats without nerve injury further supported a role of CALP2 in the regulation of TNF-α expression. Although clinical trials of calpain inhibition therapy for alleviation of neuropathic pain induced by motor nerve injury have not yet shown success, our observations linking CALP2 and TNF-α provide a framework of a systems approach based perspective for treating neuropathic pain. Copyright © 2018 IBRO. Published by Elsevier Ltd. All rights reserved.

  13. Different Roles for Contracture and Calpain in Calcium Paradox-Induced Heart Injury

    Science.gov (United States)

    Zhang, Jian-Ying; Bi, Sheng-Hui; Xu, Ming; Jin, Zhen-Xiao; Yang, Yang; Jiang, Xiao-Fan; Zhou, Jing-Jun

    2012-01-01

    The Ca2+ paradox represents a good model to study Ca2+ overload injury in ischemic heart diseases. We and others have demonstrated that contracture and calpain are involved in the Ca2+ paradox-induced injury. This study aimed to elucidate their roles in this model. The Ca2+ paradox was elicited by perfusing isolated rat hearts with Ca2+-free KH media for 3 min or 5 min followed by 30 min of Ca2+ repletion. The LVDP was measured to reflect contractile function, and the LVEDP was measured to indicate contracture. TTC staining and the quantification of LDH release were used to define cell death. Calpain activity and troponin I release were measured after Ca2+ repletion. Ca2+ repletion of the once 3-min Ca2+ depleted hearts resulted in almost no viable tissues and the disappearance of contractile function. Compared to the effects of the calpain inhibitor MDL28170, KB-R7943, an inhibitor of the Na+/Ca2+ exchanger, reduced the LVEDP level to a greater extent, which was well correlated with improved contractile function recovery and tissue survival. The depletion of Ca2+ for 5 min had the same effects on injury as the 3-min Ca2+ depletion, except that the LVEDP in the 5-min Ca2+ depletion group was lower than the level in the 3-min Ca2+ depletion group. KB-R7943 failed to reduce the level of LVEDP, with no improvement in the LVDP recovery in the hearts subjected to the 5-min Ca2+ depletion treatment; however, KB-R7943 preserved its protective effects in surviving tissue. Both KB-R7943 and MDL28170 attenuated the Ca2+ repletion-induced increase in calpain activity in 3 min or 5 min Ca2+ depleted hearts. However, only KB-R7943 reduced the release of troponin I from the Ca2+ paradoxic heart. These results provide evidence suggesting that contracture is the main cause for contractile dysfunction, while activation of calpain mediates cell death in the Ca2+ paradox. PMID:23284963

  14. Genetic disruption of calpain correlates with loss of membrane blebbing and differential expression of RhoGDI-1, cofilin and tropomyosin

    DEFF Research Database (Denmark)

    Larsen, Anna Karina; Lametsch, Rene; Elce, John S.

    2008-01-01

    blebbing was significantly reduced in calpain-knockout cells, and genetic rescue fully restored the wild-type phenotype in knockout cells. Proteomic comparison of wild-type and knockout cells identified decreased levels of RhoGDI-1 (Rho GDP-dissociation inhibitor) and cofilin 1, and increased levels...

  15. Suppression of NHE1 by small interfering RNA inhibits HIF-1α-induced angiogenesis in vitro via modulation of calpain activity.

    Science.gov (United States)

    Mo, Xian-Gang; Chen, Qing-Wei; Li, Xing-Sheng; Zheng, Min-Ming; Ke, Da-Zhi; Deng, Wei; Li, Gui-Qiong; Jiang, Jin; Wu, Zhi-Qin; Wang, Li; Wang, Peng; Yang, Yan; Cao, Guang-Yi

    2011-03-01

    Hypoxia-inducible factor-1 (HIF-1) orchestrates angiogenesis under hypoxic conditions mainly due to increased expression of such target genes as vascular endothelial growth factor (VEGF). Na+/H+exchanger-1 (NHE1), a potential HIF target gene product, plays a pivotal role in proliferation, survival, migration, adhesion and so on. However, it is unknown whether NHE1 is involved in HIF-1α-induced angiogenesis. This present study demonstrated that the expression of NHE1 was much higher in human umbilical vein endothelial cells (HUVECs) infected with adenovirus encoding HIF-1α (rAd-HIF) than with vacuum adenovirus (vAd). HIF-1α also increased the expression of VEGF, the expression and activity of calpains, and the intracellular pH. Moreover, small interfering RNA targeting NHE1 (NHE1 siRNA) dramatically decreased the expression of NHE1 and thus lowered the intracellular pH, and it also attenuated the protein expression of calpain-2 but not calpain-1, resulting in the lower calpain activity. Furthermore, HIF-1α enhanced the proliferation, migration and Matrigel tube formation, which were inhibited by NHE1 siRNA. Finally, the inhibitory effect of NHE1 siRNA was reversed by VEGF and the reversibility of the later was abrogated by the calpain inhibitor ALLM. In conclusion, the findings have revealed that NHE1 might participate in HIF-1-induced angiogenesis due, at least in part, to the alteration of the calpain activity, suggesting that NHE1 as well as calpains might represent a potential target of controlling angiogenesis in response to the hypoxic stress under various pathological conditions. Copyright © 2010 Elsevier Inc. All rights reserved.

  16. Concurrent calpain and caspase-3 mediated proteolysis of alpha II-spectrin and tau in rat brain after methamphetamine exposure: a similar profile to traumatic brain injury.

    Science.gov (United States)

    Warren, Matthew W; Kobeissy, Firas H; Liu, Ming Cheng; Hayes, Ronald L; Gold, Mark S; Wang, Kevin K W

    2005-12-05

    Neurotoxicity in rat cortex and hippocampus following acute methamphetamine administration was characterized and compared to changes following traumatic brain injury. Doses of 10, 20, and 40 mg/kg of methamphetamine produced significant increases in calpain- and caspase-cleaved alpha II-spectrin and tau protein fragments, suggesting cell injury or death. Changes in proteolytic products were significantly increased over vehicle controls. Use of fragment specific biomarkers detected prominent calpain-mediated protein fragments in the cortex and hippocampus while caspase-mediated protein fragments were also detected in the hippocampus. Remarkably, proteolytic product increases at the 40 mg/kg dose after 24 h were as high as those observed in experimental traumatic brain injury. Use of calpain and caspase proteolytic inhibitors may be useful in preventing methamphetamine-induced neurotoxicity.

  17. Propofol Ameliorates Calpain-induced Collapsin Response Mediator Protein-2 Proteolysis in Traumatic Brain Injury in Rats

    Science.gov (United States)

    Yu, Yun; Jian, Min-Yu; Wang, Yun-Zhen; Han, Ru-Quan

    2015-01-01

    Background: Collapsin response mediator protein-2 (CRMP2), a multifunctional cytosolic protein highly expressed in the brain, is degraded by calpain following traumatic brain injury (TBI), possibly inhibiting posttraumatic neurite regeneration. Lipid peroxidation (LP) is involved in triggering postinjury CRMP2 proteolysis. We examined the hypothesis that propofol could attenuate LP, calpain-induced CRMP2 degradation, and brain injury after TBI. Methods: A unilateral moderate controlled cortical impact injury was induced in adult male Sprague-Dawley rats. The animals were randomly divided into seven groups: Sham control group, TBI group, TBI + propofol groups (including propofol 1 h, 2 h, and 4 h groups), TBI + U83836E group and TBI + fat emulsion group. The LP inhibitor U83836E was used as a control to identify that antioxidation partially accounts for the potential neuroprotective effects of propofol. The solvent of propofol, fat emulsion, was used as the vehicle control. Ipsilateral cortex tissues were harvested at 24 h post-TBI. Immunofluorescent staining, Western blot analysis, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling were used to evaluate LP, calpain activity, CRMP2 proteolysis and programmed cell death. The data were statistically analyzed using one-way analysis of variance and a paired t-test. Results: Propofol and U83836E significantly ameliorated the CRMP2 proteolysis. In addition, both propofol and U83836E significantly decreased the ratio of 145-kDa αII-spectrin breakdown products to intact 270-kDa spectrin, the 4-hydroxynonenal expression and programmed cell death in the pericontusional cortex at 24 h after TBI. There was no difference between the TBI group and the fat emulsion group. Conclusions: These results demonstrate that propofol postconditioning alleviates calpain-mediated CRMP2 proteolysis and provides neuroprotective effects following moderate TBI potentially by counteracting LP and reducing calpain activation

  18. Calpain-3 impairs cell proliferation and stimulates oxidative stress-mediated cell death in melanoma cells.

    Directory of Open Access Journals (Sweden)

    Daniele Moretti

    Full Text Available Calpain-3 is an intracellular cysteine protease, belonging to Calpain superfamily and predominantly expressed in skeletal muscle. In human melanoma cell lines and biopsies, we previously identified two novel splicing variants (hMp78 and hMp84 of Calpain-3 gene (CAPN3, which have a significant lower expression in vertical growth phase melanomas and, even lower, in metastases, compared to benign nevi. In the present study, in order to investigate the pathophysiological role played by the longer Calpain-3 variant, hMp84, in melanoma cells, we over-expressed it in A375 and HT-144 cells. In A375 cells, the enforced expression of hMp84 induces p53 stabilization, and modulates the expression of a few p53- and oxidative stress-related genes. Consistently, hMp84 increases the intracellular production of ROS (Reactive Oxygen Species, which lead to oxidative modification of phospholipids (formation of F2-isoprostanes and DNA damage. Such events culminate in an adverse cell fate, as indicated by the decrease of cell proliferation and by cell death. To a different extent, either the antioxidant N-acetyl-cysteine or the p53 inhibitor, Pifithrin-α, recover cell viability and decrease ROS formation. Similarly to A375 cells, hMp84 over-expression causes inhibition of cell proliferation, cell death, and increase of both ROS levels and F2-isoprostanes also in HT-144 cells. However, in these cells no p53 accumulation occurs. In both cell lines, no significant change of cell proliferation and cell damage is observed in cells over-expressing the mutant hMp84C42S devoid of its enzymatic activity, suggesting that the catalytic activity of hMp84 is required for its detrimental effects. Since a more aggressive phenotype is expected to benefit from down-regulation of mechanisms impairing cell growth and survival, we envisage that Calpain-3 down-regulation can be regarded as a novel mechanism contributing to melanoma progression.

  19. Calpain-mediated cleavage of DARPP-32 in Alzheimer’s disease

    Science.gov (United States)

    Cho, Kwangmin; Cho, Mi-Hyang; Seo, Jung-Han; Peak, Jongjin; Kong, Kyoung-Hye; Yoon, Seung-Yong; Kim, Dong-Hou

    2015-01-01

    Toxicity induced by aberrant protein aggregates in Alzheimer’s disease (AD) causes synaptic disconnection and concomitant progressive neurodegeneration that eventually impair cognitive function. cAMP-response element-binding protein (CREB) is a transcription factor involved in the molecular switch that converts short-term to long-term memory. Although disturbances in CREB function have been suggested to cause memory deficits in both AD and AD animal models, the mechanism of CREB dysfunction is still unclear. Here, we show that the dopamine- and cAMP-regulated phosphoprotein 32 kDa (DARPP-32), a key inhibitor of protein phosphate-1 (PP-1) that regulates CREB phosphorylation, is cleaved by activated calpain in both AD brains and neuronal cells treated with amyloid-β or okadaic acid, a protein phosphatase-2A inhibitor that induces tau hyperphosphorylation and neuronal death. We found that DARPP-32 is mainly cleaved at Thr153 by calpain and that this cleavage of DARPP-32 reduces CREB phosphorylation via loss of its inhibitory function on PP1. Our results suggest a novel mechanism of DARPP-32–CREB signalling dysregulation in AD. PMID:26178297

  20. A calcium- and calpain-dependent pathway determines the response to lenalidomide in myelodysplastic syndromes

    Science.gov (United States)

    Fang, Jing; Liu, Xiaona; Bolanos, Lyndsey; Barker, Brenden; Rigolino, Carmela; Cortelezzi, Agostino; Oliva, Esther N; Cuzzola, Maria; Grimes, H Leighton; Fontanillo, Celia; Komurov, Kakajan; MacBeth, Kyle; Starczynowski, Daniel T

    2017-01-01

    Despite the high response rates of individuals with myelodysplastic syndrome (MDS) with deletion of chromosome 5q (del(5q)) to treatment with lenalidomide (LEN) and the recent identification of cereblon (CRBN) as the molecular target of LEN, the cellular mechanism by which LEN eliminates MDS clones remains elusive. Here we performed an RNA interference screen to delineate gene regulatory networks that mediate LEN responsiveness in an MDS cell line, MDSL. We identified GPR68, which encodes a G-protein-coupled receptor that has been implicated in calcium metabolism, as the top candidate gene for modulating sensitivity to LEN. LEN induced GPR68 expression via IKAROS family zinc finger 1 (IKZF1), resulting in increased cytosolic calcium levels and activation of a calcium-dependent calpain, CAPN1, which were requisite steps for induction of apoptosis in MDS cells and in acute myeloid leukemia (AML) cells. In contrast, deletion of GPR68 or inhibition of calcium and calpain activation suppressed LEN-induced cytotoxicity. Moreover, expression of calpastatin (CAST), an endogenous CAPN1 inhibitor that is encoded by a gene (CAST) deleted in del(5q) MDS, correlated with LEN responsiveness in patients with del(5q) MDS. Depletion of CAST restored responsiveness of LEN-resistant non-del(5q) MDS cells and AML cells, providing an explanation for the superior responses of patients with del(5q) MDS to LEN treatment. Our study describes a cellular mechanism by which LEN, acting through CRBN and IKZF1, has cytotoxic effects in MDS and AML that depend on a calcium- and calpain-dependent pathway. PMID:27294874

  1. Calpain- and caspase-mediated alphaII-spectrin and tau proteolysis in rat cerebrocortical neuronal cultures after ecstasy or methamphetamine exposure.

    Science.gov (United States)

    Warren, Matthew W; Zheng, Wenrong; Kobeissy, Firas H; Cheng Liu, Ming; Hayes, Ronald L; Gold, Mark S; Larner, Stephen F; Wang, Kevin K W

    2007-08-01

    Abuse of 3,4-methylenedioxymethamphetamine (MDMA or Ecstasy) and methamphetamine (Meth or Speed) is a growing international problem with an estimated 250 million users of psychoactive drugs worldwide. It is important to demonstrate and understand the mechanism of neurotoxicity so potential prevention and treatment therapies can be designed. In this study rat primary cerebrocortical neuron cultures were challenged with MDMA and Meth (1 or 2 mM) for 24 and 48 h and compared to the excitotoxin N-methyl-D-aspartate (NMDA). The neurotoxicity of these drugs, as assessed by microscopy, lactate dehydrogenase release and immunoblot, was shown to be both dose- and time-dependent. Immunoblot analysis using biomarkers of cell death showed significant proteolysis of both alphaII-spectrin and tau proteins. Breakdown products of alphaII-spectrin (SBDPs) of 150, 145, and 120 kDa and tau breakdown products (TBDPs) of 45, 32, 26, and 14 kDa were observed. The use of the protease inhibitors calpain inhibitor SJA6017 and caspase inhibitors z-VAD-fmk and Z-D-DCB, attenuated drug-induced alphaII-spectrin and tau proteolysis. The calpain inhibitor reduced the calpain-induced breakdown products SBDP145 and TBDP14, but there was an offset increase in the caspase-mediated breakdown products SBDP120 and TBDP45. The caspase inhibitors, on the other hand, decreased SBDP120 and TBDP45. These data suggest that both MDMA and Meth trigger concerted proteolytic attacks of the structural proteins by both calpain and caspase family of proteases. The ability of the protease inhibitors to reduce the damage caused by these drugs suggests that the treatment arsenal could include similar drugs as possible tools to combat the drug-induced neurotoxicity in vivo.

  2. Modulation of intracellular calcium levels by calcium lactate affects colon cancer cell motility through calcium-dependent calpain.

    Science.gov (United States)

    Sundaramoorthy, Pasupathi; Sim, Jae Jun; Jang, Yeong-Su; Mishra, Siddhartha Kumar; Jeong, Keun-Yeong; Mander, Poonam; Chul, Oh Byung; Shim, Won-Sik; Oh, Seung Hyun; Nam, Ky-Youb; Kim, Hwan Mook

    2015-01-01

    Cancer cell motility is a key phenomenon regulating invasion and metastasis. Focal adhesion kinase (FAK) plays a major role in cellular adhesion and metastasis of various cancers. The relationship between dietary supplementation of calcium and colon cancer has been extensively investigated. However, the effect of calcium (Ca2+) supplementation on calpain-FAK-motility is not clearly understood. We sought to identify the mechanism of FAK cleavage through Ca2+ bound lactate (CaLa), its downstream signaling and role in the motility of human colon cancer cells. We found that treating HCT116 and HT-29 cells with CaLa immediately increased the intracellular Ca2+ (iCa2+) levels for a prolonged period of time. Ca2+ influx induced cleavage of FAK into an N-terminal FAK (FERM domain) in a dose-dependent manner. Phosphorylated FAK (p-FAK) was also cleaved in to its p-N-terminal FAK. CaLa increased colon cancer cells motility. Calpeptin, a calpain inhibitor, reversed the effects of CaLa on FAK and pFAK cleavage in both cancer cell lines. The cleaved FAK translocates into the nucleus and modulates p53 stability through MDM2-associated ubiquitination. CaLa-induced Ca2+ influx increased the motility of colon cancer cells was mediated by calpain activity through FAK and pFAK protein destabilization. In conclusion, these results suggest that careful consideration may be given in deciding dietary Ca2+ supplementation to patient undergoing treatment for metastatic cancer.

  3. Decreased calpain activity in chronic myeloid leukemia impairs apoptosis by increasing survivin in myeloid progenitors and xiap1 in differentiating granulocytes

    Science.gov (United States)

    Huang, Weiqi; Bei, Ling; Hjort, Elizabeth E.; Eklund, Elizabeth A.

    2017-01-01

    Chronic Myeloid Leukemia (CML) is characterized by translocations between chromosomes 9 and 22, resulting in expression of Bcr-abl oncogenes. Although the clinical course of CML was revolutionized by development of Bcr-abl-directed tyrosine kinase inhibitors (TKIs), CML is not cured by these agents. Specifically, the majority of subjects relapsed in clinical trials attempting TKI discontinuation, suggesting persistence of leukemia stem cells (LSCs) even in molecular remission. Identifying mechanisms of CML-LSC persistence may suggest rationale therapeutic targets to augment TKI efficacy and lead to cure. Apoptosis resistance is one proposed mechanism. In prior studies, we identified increased expression of Growth Arrest Specific 2 (Gas2; a Calpain inhibitor) in Bcr-abl+ bone marrow progenitor cells. A number of previously described Calpain substrates might influence apoptosis in CML, including βcatenin and the X-linked Inhibitor of Apoptosis Protein 1 (Xiap1). We previously found Gas2/Calpain dependent stabilization of βcatenin in CML, and increased expression of βcatenin target genes, including Survivin (also an IAP). In the current work, we investigate contributions of Survivin and Xiap1 to Fas-resistance in Bcr-abl+ bone marrow cells. Inhibitors of these proteins are currently in clinical trials for other malignancies, but a role for either IAP in CML-LSC persistence is unknown. PMID:28881589

  4. Calpain 3 is a rapid-action, unidirectional proteolytic switch central to muscle remodeling.

    Directory of Open Access Journals (Sweden)

    Antoine de Morrée

    Full Text Available Calpain 3 (CAPN3 is a cysteine protease that when mutated causes Limb Girdle Muscular Dystrophy 2A. It is thereby the only described Calpain family member that genetically causes a disease. Due to its inherent instability little is known of its substrates or its mechanism of activity and pathogenicity. In this investigation we define a primary sequence motif underlying CAPN3 substrate cleavage. This motif can transform non-related proteins into substrates, and identifies >300 new putative CAPN3 targets. Bioinformatic analyses of these targets demonstrate a critical role in muscle cytoskeletal remodeling and identify novel CAPN3 functions. Among the new CAPN3 substrates are three E3 SUMO ligases of the Protein Inhibitor of Activated Stats (PIAS family. CAPN3 can cleave PIAS proteins and negatively regulates PIAS3 sumoylase activity. Consequently, SUMO2 is deregulated in patient muscle tissue. Our study thus uncovers unexpected crosstalk between CAPN3 proteolysis and protein sumoylation, with strong implications for muscle remodeling.

  5. Brucella infection inhibits macrophages apoptosis via Nedd4-dependent degradation of calpain2.

    Science.gov (United States)

    Cui, Guimei; Wei, Pan; Zhao, Yuxi; Guan, Zhenhong; Yang, Li; Sun, Wanchun; Wang, Shuangxi; Peng, Qisheng

    2014-11-07

    The calcium-dependent protease calpain2 is involved in macrophages apoptosis. Brucella infection-induced up-regulation of intracellular calcium level is an essential factor for the intracellular survival of Brucella within macrophages. Here, we hypothesize that calcium-dependent E3 ubiquitin ligase Nedd4 ubiquitinates calpain2 and inhibits Brucella infection-induced macrophage apoptosis via degradation of calpain2.Our results reveal that Brucella infection induces increases in Nedd4 activity in an intracellular calcium dependent manner. Furthermore, Brucella infection-induced degradation of calpain2 is mediated by Nedd4 ubiquitination of calpain2. Brucella infection-induced calpain2 degradation inhibited macrophages apoptosis. Treatment of Brucella infected macrophages with calcium chelator BAPTA or Nedd4 knock-down decreased Nedd4 activity, prevented calpain2 degradation, and resulted in macrophages apoptosis. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Vital role of the calpain-calpastatin system for placental-integrity-dependent embryonic survival.

    Science.gov (United States)

    Takano, Jiro; Mihira, Naomi; Fujioka, Ryo; Hosoki, Emi; Chishti, Athar H; Saido, Takaomi C

    2011-10-01

    Although the calpain-calpastatin system has been implicated in a number of pathological conditions, its normal physiological role remains largely unknown. To investigate the functions of this system, we generated conventional and conditional calpain-2 knockout mice. The conventional calpain-2 knockout embryos died around embryonic day 15, preceded by cell death associated with caspase activation and DNA fragmentation in placental trophoblasts. In contrast, conditional knockout mice in which calpain-2 is expressed in the placenta but not in the fetus were spared. These results suggest that calpain-2 contributes to trophoblast survival via suppression of caspase activation. Double-knockout mice also deficient in calpain-1 and calpastatin resulted in accelerated and rescued embryonic lethality, respectively, suggesting that calpain-1 and -2 at least in part share similar in vivo functions under the control of calpastatin. Triple-knockout mice exhibited early embryonic lethality, a finding consistent with the notion that this protease system is vital for embryonic survival.

  7. SwissProt search result: AK109961 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK109961 002-159-B07 (P30403) Hemorrhagic protein-rhodostomin precursor (EC 3.4.24....-) (RHO) [Contains: Disintegrin rhodostomin (Disintegrin kistrin) (Platelet aggregation activation inhibitor)] DISR_AGKRH 5e-12 ...

  8. SwissProt search result: AK106798 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK106798 002-116-B08 (P02873) Alpha-amylase inhibitor 1 precursor (Alpha-AI-1) (Alp...ha-AI1) (Lectin) [Contains: Alpha-amylase inhibitor 1 chain 1; Alpha-amylase inhibitor 1 chain 2] LEA1_PHAVU 1e-19 ...

  9. Reduced syncytin-1 expression in choriocarcinoma BeWo cells activates the calpain1-AIF-mediated apoptosis, implication for preeclampsia.

    Science.gov (United States)

    Huang, Qiang; Chen, Haibin; Wang, Fengchao; Brost, Brian C; Li, Jinping; Gao, Yu; Li, Zongfang; Gao, Ya; Jiang, Shi-Wen

    2014-08-01

    Placentas associated with preeclampsia are characterized by extensive apoptosis in trophoblast lineages. Syncytin-1 (HERVWE1) mediates the fusion of cytotrophoblasts to form syncytiotrophoblasts, which assume the placental barrier, fetal-maternal exchange and endocrine functions. While decreased syncytin-1 expression has been observed in preeclamptic placentas, it is not clear if this alteration is involved in trophoblast apoptosis. In the current study, we found that siRNA-mediated knockdown of syncytin-1 led to apoptosis in choriocarcinoma BeWo, a cell line of trophoblastic origin. Characterization of the apoptotic pathways indicated that this effect does not rely on the activation of caspases. Rather, decreased syncytin-1 levels activated the apoptosis inducing factor (AIF) apoptotic pathway by inducing the expression, cleavage, and nuclear translocation of AIF. Moreover, calpain1, the cysteine protease capable of cleaving AIF, was upregulated by syncytin-1 knockdown. Furthermore, treatment with calpain1 inhibitor MDL28170 effectively reversed AIF cleavage, AIF nuclear translocation, and cell apoptosis triggered by syncytin-1 downregulation, verifying the specific action of calpain1-AIF pathway in trophoblast apoptosis. We confirmed that preeclamptic placentas express lower levels of syncytin-1 than normal placentas, and observed an inverse correlation between syncytin-1 and AIF/calpain1 mRNA levels, a result consistent with the in vitro findings. Immunohistochemistry analyses indicated decreased syncytin-1 and increased AIF and calpain1 protein levels in apoptotic cells of preeclamptic placentas. These findings have for the first time revealed that decreased levels of syncytin-1 can trigger the AIF-mediated apoptosis pathway in BeWo cells. This novel mechanism may contribute to the structural and functional deficiencies of syncytium frequently observed in preeclamptic placentas.

  10. Reduced syncytin-1 expression in chriocarcinoma BeWo cells activates the calpain1-AIF-mediated apoptosis, implication for preeclampsia

    Science.gov (United States)

    Huang, Qiang; Chen, Haibin; Wang, Fengchao; Brost, Brian C.; Li, Jinping; Gao, Yu; Li, Zongfang; Gao, Ya; Jiang, Shi-Wen

    2015-01-01

    Placentas associated with preeclampsia are characterized by extensive apoptosis in trophoblast lineages. Syncytin-1 (HERVWE1) mediates the fusion of cytotrophoblasts to form syncytiotrophoblasts, which assume the placental barrier, fetal-maternal exchange and endocrine functions. While decreased syncytin-1 expression has been observed in preeclamptic placentas, it is not clear if this alteration is involved in trophoblast apoptosis. In the current study we found that siRNA-mediated knockdown of syncytin-1 led to apoptosis in choriocarcinoma BeWo, a cell line of trophoblastic origin. Characterization of the apoptotic pathways indicated that this effect does not rely on the activation of caspases. Rather, decreased syncytin-1 levels activated the AIF apoptotic pathway by inducing the expression, cleavage, and nuclear translocation of AIF. Moreover, calpain1, the cysteine protease capable of cleaving AIF, was upregulated by syncytin-1 knockdown. Furthermore, treatment with calpain1 inhibitor MDL28170 effectively reversed AIF cleavage, AIF nuclear translocation, and cell apoptosis triggered by syncytin-1 downregulation, verifying the specific action of calpain1-AIF pathway in trophoblast apoptosis. We confirmed that preeclamptic placentas express lower levels of syncytin-1 than normal placentas, and observed an inverse correlation between syncytin-1 and AIF/calpain1 mRNA levels, a result consistent with the in vitro findings. Immunohistochemistry analyses indicated decreased syncytin-1, increased AIF and calpain1 protein levels in apoptotic cells of preeclamptic placentas. These findings have for the first time revealed that decreased levels of syncytin-1 can trigger the AIF-mediated apoptosis pathway in BeWo cells. This novel mechanism may contribute to the structural and functional deficiencies of syncytium frequently observed in preeclamptic placentas. PMID:24413738

  11. Calpain 3 is important for muscle regeneration: Evidence from patients with limb girdle muscular dystrophies

    Science.gov (United States)

    2012-01-01

    Background Limb girdle muscular dystrophy (LGMD) type 2A is caused by mutations in the CAPN3 gene and complete lack of functional calpain 3 leads to the most severe muscle wasting. Calpain 3 is suggested to be involved in maturation of contractile elements after muscle degeneration. The aim of this study was to investigate how mutations in the four functional domains of calpain 3 affect muscle regeneration. Methods We studied muscle regeneration in 22 patients with LGMD2A with calpain 3 deficiency, in five patients with LGMD2I, with a secondary reduction in calpain 3, and in five patients with Becker muscular dystrophy (BMD) with normal calpain 3 levels. Regeneration was assessed by using the developmental markers neonatal myosin heavy chain (nMHC), vimentin, MyoD and myogenin and counting internally nucleated fibers. Results We found that the recent regeneration as determined by the number of nMHC/vimentin-positive fibers was greatly diminished in severely affected LGMD2A patients compared to similarly affected patients with LGMD2I and BMD. Whorled fibers, a sign of aberrant regeneration, was highly elevated in patients with a complete lack of calpain 3 compared to patients with residual calpain 3. Regeneration is not affected by location of the mutation in the CAPN3 gene. Conclusions Our findings suggest that calpain 3 is needed for the regenerative process probably during sarcomere remodeling as the complete lack of functional calpain 3 leads to the most severe phenotypes. PMID:22443334

  12. Calpain 3 is important for muscle regeneration: Evidence from patients with limb girdle muscular dystrophies

    Directory of Open Access Journals (Sweden)

    Hauerslev Simon

    2012-03-01

    Full Text Available Abstract Background Limb girdle muscular dystrophy (LGMD type 2A is caused by mutations in the CAPN3 gene and complete lack of functional calpain 3 leads to the most severe muscle wasting. Calpain 3 is suggested to be involved in maturation of contractile elements after muscle degeneration. The aim of this study was to investigate how mutations in the four functional domains of calpain 3 affect muscle regeneration. Methods We studied muscle regeneration in 22 patients with LGMD2A with calpain 3 deficiency, in five patients with LGMD2I, with a secondary reduction in calpain 3, and in five patients with Becker muscular dystrophy (BMD with normal calpain 3 levels. Regeneration was assessed by using the developmental markers neonatal myosin heavy chain (nMHC, vimentin, MyoD and myogenin and counting internally nucleated fibers. Results We found that the recent regeneration as determined by the number of nMHC/vimentin-positive fibers was greatly diminished in severely affected LGMD2A patients compared to similarly affected patients with LGMD2I and BMD. Whorled fibers, a sign of aberrant regeneration, was highly elevated in patients with a complete lack of calpain 3 compared to patients with residual calpain 3. Regeneration is not affected by location of the mutation in the CAPN3 gene. Conclusions Our findings suggest that calpain 3 is needed for the regenerative process probably during sarcomere remodeling as the complete lack of functional calpain 3 leads to the most severe phenotypes.

  13. Calpain-2-mediated PTEN degradation contributes to BDNF-induced stimulation of dendritic protein synthesis

    Science.gov (United States)

    Briz, Victor; Hsu, Yu-Tien; Li, Yi; Lee, Erin; Bi, Xiaoning; Baudry, Michel

    2013-01-01

    Memory consolidation has been suggested to be protein synthesis-dependent. Recent data indicate that BDNF-induced dendritic protein synthesis is a key event in memory formation through activation of the mammalian target of rapamycin (mTOR) pathway. BDNF also activates calpain, a calcium-dependent cysteine protease, which has been shown to play a critical role in learning and memory. This study was therefore directed at testing the hypothesis that calpain activity is required for BDNF-stimulated local protein synthesis, and at identifying the underlying molecular mechanism. In rat hippocampal slices, cortical synaptoneurosomes, and cultured neurons, BDNF-induced mTOR pathway activation and protein translation were blocked by calpain inhibition. BDNF treatment rapidly reduced levels of hamartin and tuberin, negative regulators of mTOR, in a calpain-dependent manner. Treatment of brain homogenates with purified calpain-1 and calpain-2 truncated both proteins. BDNF treatment increased phosphorylation of both Akt and ERK, but only the effect on Akt was blocked by calpain inhibition. Levels of PTEN (phosphatase and tensin homolog deleted on chromosome ten), a phosphatase that inactivates Akt, were decreased following BDNF treatment, and calpain inhibition reversed this effect. Calpain-2 but not calpain-1 treatment of brain homogenates resulted in PTEN degradation. In cultured cortical neurons, knock-down of calpain-2 but not calpain-1 by siRNA completely suppressed the effect of BDNF on mTOR activation. Our results reveal a critical role for calpain-2 in BDNF-induced mTOR signaling and dendritic protein synthesis via PTEN, hamartin and tuberin degradation. This mechanism therefore provides a link between proteolysis and protein synthesis that might contribute to synaptic plasticity. PMID:23467348

  14. Arabidopsis CDS blastp result: AK101251 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101251 J033032F02 At3g14310.1 pectinesterase family protein contains Pfam profiles: PF01095 pectin...esterase, PF04043 plant invertase/pectin methylesterase inhibitor ;similar to pectin methylesterase GB:Q42534 from [Arabidopsis thaliana] 1e-129 ...

  15. m-Calpain is required for preimplantation embryonic development in mice

    Directory of Open Access Journals (Sweden)

    Williams Karen

    2006-01-01

    Full Text Available Abstract Background μ-calpain and m-calpain are ubiquitously expressed proteases implicated in cellular migration, cell cycle progression, degenerative processes and cell death. These heterodimeric enzymes are composed of distinct catalytic subunits, encoded by Capn1 (μ-calpain or Capn2 (m-calpain, and a common regulatory subunit encoded by Capn4. Disruption of the mouse Capn4 gene abolished both μ-calpain and m-calpain activity, and resulted in embryonic lethality, thereby suggesting essential roles for one or both of these enzymes during mammalian embryogenesis. Disruption of the Capn1 gene produced viable, fertile mice implying that either m-calpain could compensate for the loss of μ-calpain, or that the loss of m-calpain was responsible for death of Capn4-/- mice. Results To distinguish between the alternatives described above, we deleted an essential coding region in the mouse Capn2 gene in embryonic stems cells and transmitted this mutant allele through the mouse germline. Breeding of heterozygous animals failed to produce homozygous mutant live offspring or implanted embryos. A nested PCR genotyping protocol was established, and homozygous preimplantation mutant embryos were detected at the morula but not at the blastocyts stage. Conclusion We conclude that homozygous disruption of the Capn2 gene results in pre-implantation embryonic lethality between the morula and blastocyst stage. This establishes that μ-calpain and m-calpain have distinct functions, and that m-calpain is vital for development of the preimplantation murine embryo.

  16. Characterization of the definitive classical calpain family of vertebrates using phylogenetic, evolutionary and expression analyses

    Science.gov (United States)

    Macqueen, Daniel J.; Wilcox, Alexander H.

    2014-01-01

    The calpains are a superfamily of proteases with extensive relevance to human health and welfare. Vast research attention is given to the vertebrate ‘classical’ subfamily, making it surprising that the evolutionary origins, distribution and relationships of these genes is poorly characterized. Consequently, there exists uncertainty about the conservation of gene family structure, function and expression that has been principally defined from work with mammals. Here, more than 200 vertebrate classical calpains were incorporated in phylogenetic analyses spanning an unprecedented range of taxa, including jawless and cartilaginous fish. We demonstrate that the common vertebrate ancestor had at least six classical calpains, including a single gene that gave rise to CAPN11, 1, 2 and 8 in the early jawed fish lineage, plus CAPN3, 9, 12, 13 and a novel calpain gene, hereafter named CAPN17. We reveal that while all vertebrate classical calpains have been subject to persistent purifying selection during evolution, the degree and nature of selective pressure has often been lineage-dependent. The tissue expression of the complete classic calpain family was assessed in representative teleost fish, amphibians, reptiles and mammals. This highlighted systematic divergence in expression across vertebrate taxa, with most classic calpain genes from fish and amphibians having more extensive tissue distribution than in amniotes. Our data suggest that classical calpain functions have frequently diverged during vertebrate evolution and challenge the ongoing value of the established system of classifying calpains by expression. PMID:24718597

  17. Calpain2 mediates Rab5-driven focal adhesion disassembly and cell migration.

    Science.gov (United States)

    Mendoza, Pablo A; Silva, Patricio; Díaz, Jorge; Arriagada, Cecilia; Canales, Jimena; Cerda, Oscar; Torres, Vicente A

    2017-11-03

    The early endosome protein Rab5 was recently shown to promote cell migration by enhancing focal adhesion disassembly through mechanisms that remain elusive. Focal adhesion disassembly is associated to proteolysis of talin, in a process that requires calpain2. Since calpain2 has been found at vesicles and endosomal compartments, we hypothesized that Rab5 stimulates calpain2 activity, leading to enhanced focal adhesion disassembly in migrating cells. We observed that calpain2 co-localizes with EEA1-positive early endosomes and co-immunoprecipitates with EEA1 and Rab5 in A549 lung carcinoma cells undergoing spreading, whereas Rab5 knock-down decreased the accumulation of calpain2 at early endosomal-enriched fractions. In addition, Rab5 silencing decreased calpain2 activity, as shown by cleavage of the fluorogenic substrate tBOC-LM-CMAC and the endogenous substrate talin. Accordingly, Rab5 promoted focal adhesion disassembly in a calpain2-dependent manner, as expression of GFP-Rab5 accelerated focal adhesion disassembly in nocodazole-synchronized cells, whereas pharmacological inhibition of calpain2 with N-acetyl-Leu-Leu-Met prevented both focal adhesion disassembly and cell migration induced by Rab5. In summary, these data uncover Rab5 as a novel regulator of calpain2 activity and focal adhesion proteolysis leading to cell migration.

  18. Asymmetric Localization of Calpain 2 during Neutrophil Chemotaxis

    OpenAIRE

    Nuzzi, Paul A.; Senetar, Melissa A.; Huttenlocher, Anna

    2007-01-01

    Chemoattractants induce neutrophil polarization through localized polymerization of F-actin at the leading edge. The suppression of rear and lateral protrusions is required for efficient chemotaxis and involves the temporal and spatial segregation of signaling molecules. We have previously shown that the intracellular calcium-dependent protease calpain is required for cell migration and is involved in regulating neutrophil chemotaxis. Here, we show that primary neutrophils and neutrophil-like...

  19. Cleavage of desmin by cysteine proteases: Calpains and cathepsin B

    DEFF Research Database (Denmark)

    Baron, Caroline; Jacobsen, S.; Purslow, P.P.

    2004-01-01

    a sequential C-terminal degradation pattern characteristic of this dipeptylpeptidase. The substrate primary structure was not found to be essential for regulation of the proteolytic activity of the cysteine peptidases studied. However, the degradation patterns obtained imply that calpains are involved...... in degradation of desmin early post-mortem, targeting the non-helical region of the desmin molecule and resulting in depolymerisation and initial disorganisation of the intermediate filament structures of the muscle cell....

  20. Pharmacological inhibition of caspase and calpain proteases: a novel strategy to enhance the homing responses of cord blood HSPCs during expansion.

    Directory of Open Access Journals (Sweden)

    V M Sangeetha

    Full Text Available BACKGROUND: Expansion of hematopoietic stem/progenitor cells (HSPCs is a well-known strategy employed to facilitate the transplantation outcome. We have previously shown that the prevention of apoptosis by the inhibition of cysteine proteases, caspase and calpain played an important role in the expansion and engraftment of cord blood (CB derived HSPCs. We hypothesize that these protease inhibitors might have maneuvered the adhesive and migratory properties of the cells rendering them to be retained in the bone marrow for sustained engraftment. The current study was aimed to investigate the mechanism of the homing responses of CB cells during expansion. METHODOLOGY/PRINCIPAL FINDINGS: CB derived CD34(+ cells were expanded using a combination of growth factors with and without Caspase inhibitor -zVADfmk or Calpain 1 inhibitor- zLLYfmk. The cells were analyzed for the expression of homing-related molecules. In vitro adhesive/migratory interactions and actin polymerization dynamics of HSPCs were assessed. In vivo homing assays were carried out in NOD/SCID mice to corroborate these observations. We observed that the presence of zVADfmk or zLLYfmk (inhibitors caused the functional up regulation of CXCR4, integrins, and adhesion molecules, reflecting in a higher migration and adhesive interactions in vitro. The enhanced actin polymerization and the RhoGTPase protein expression complemented these observations. Furthermore, in vivo experiments showed a significantly enhanced homing to the bone marrow of NOD/SCID mice. CONCLUSION/SIGNIFICANCE: Our present study reveals another novel aspect of the regulation of caspase and calpain proteases in the biology of HSPCs. The priming of the homing responses of the inhibitor-cultured HSPCs compared to the cytokine-graft suggests that the modulation of these proteases may help in overcoming the major homing defects prevalent in the expansion cultures thereby facilitating the manipulation of cells for transplant

  1. Muscle pathology in 31 patients with calpain 3 gene mutations..

    Science.gov (United States)

    Nadaj-Pakleza, Aleksandra A; Dorobek, M; Nestorowicz, K; Ryniewicz, B; Szmidt-Sałkowska, E; Kamińska, A M

    2013-01-01

    At present, more than 20 different forms of limb-girdle muscular dystrophies (LGMDs) are known (at least 7 autosomal dominant and 14 autosomal recessive). Although these different forms show some typical phenotypic characteristics, the existing clinical overlap makes their differential diagnosis difficult. Limb-girdle muscular dystrophy type 2 (LGMD2A) is the most prevalent LGMD in many European as well as Brazilian communities and is caused by mutations in the gene CAPN3. Laboratory testing, such as calpain immunohistochemistry and Western-blot analysis, is not totally reliable, since up to 20% of molecularly confirmed LGMD2A show normal content of calpain 3 and a third of LGMD2A biopsies have normal calpain 3 proteo-lytic activity in the muscle. Thus, genetic testing is considered as the only reliable diagnostic criterion in LGMD2A. In an attempt to find a correlation between genotype and muscle pathology in limb-girdle muscular dystrophy 2A we performed histopathological investigation of a group of 31 patients subdivided according to the type of pathologic CAPN3 gene mutation. In all biopsies typical features of muscular dystrophy such as fiber necrosis and regeneration, variation in fiber size and fibrosis were noted. Lobulated fibers were often encountered in the muscle biopsies of LGMD2A patients. Such fibers were more frequent in patients with 550delA mutation. These findings may be helpful in establishing diagnostic strategies in LGMD.

  2. Inhibition of Starvation-Triggered Endoplasmic Reticulum Stress, Autophagy, and Apoptosis in ARPE-19 Cells by Taurine through Modulating the Expression of Calpain-1 and Calpain-2.

    Science.gov (United States)

    Zhang, Yuanyuan; Ren, Shu; Liu, Yuci; Gao, Kun; Liu, Zheng; Zhang, Zhou

    2017-10-14

    Age-related macular degeneration (AMD) is a complex disease with multiple initiators and pathways that converge on death for retinal pigment epithelial (RPE) cells. In this study, effects of taurine on calpains, autophagy, endoplasmic reticulum (ER) stress, and apoptosis in ARPE-19 cells (a human RPE cell line) were investigated. We first confirmed that autophagy, ER stress and apoptosis in ARPE-19 cells were induced by Earle's balanced salt solution (EBSS) through starvation to induce RPE metabolic stress. Secondly, inhibition of ER stress by 4-phenyl butyric acid (4-PBA) alleviated autophagy and apoptosis, and suppression of autophagy by 3-methyl adenine (3-MA) reduced the cell apoptosis, but the ER stress was minimally affected. Thirdly, the apoptosis, ER stress and autophagy were inhibited by gene silencing of calpain-2 and overexpression of calpain-1, respectively. Finally, taurine suppressed both the changes of the important upstream regulators (calpain-1 and calpain-2) and the activation of ER stress, autophagy and apoptosis, and taurine had protective effects on the survival of ARPE-19 cells. Collectively, this data indicate that taurine inhibits starvation-triggered endoplasmic reticulum stress, autophagy, and apoptosis in ARPE-19 cells by modulating the expression of calpain-1 and calpain-2.

  3. Inhibition of Starvation-Triggered Endoplasmic Reticulum Stress, Autophagy, and Apoptosis in ARPE-19 Cells by Taurine through Modulating the Expression of Calpain-1 and Calpain-2

    Directory of Open Access Journals (Sweden)

    Yuanyuan Zhang

    2017-10-01

    Full Text Available Age-related macular degeneration (AMD is a complex disease with multiple initiators and pathways that converge on death for retinal pigment epithelial (RPE cells. In this study, effects of taurine on calpains, autophagy, endoplasmic reticulum (ER stress, and apoptosis in ARPE-19 cells (a human RPE cell line were investigated. We first confirmed that autophagy, ER stress and apoptosis in ARPE-19 cells were induced by Earle’s balanced salt solution (EBSS through starvation to induce RPE metabolic stress. Secondly, inhibition of ER stress by 4-phenyl butyric acid (4-PBA alleviated autophagy and apoptosis, and suppression of autophagy by 3-methyl adenine (3-MA reduced the cell apoptosis, but the ER stress was minimally affected. Thirdly, the apoptosis, ER stress and autophagy were inhibited by gene silencing of calpain-2 and overexpression of calpain-1, respectively. Finally, taurine suppressed both the changes of the important upstream regulators (calpain-1 and calpain-2 and the activation of ER stress, autophagy and apoptosis, and taurine had protective effects on the survival of ARPE-19 cells. Collectively, this data indicate that taurine inhibits starvation-triggered endoplasmic reticulum stress, autophagy, and apoptosis in ARPE-19 cells by modulating the expression of calpain-1 and calpain-2.

  4. Modulation of intracellular calcium levels by calcium lactate affects colon cancer cell motility through calcium-dependent calpain.

    Directory of Open Access Journals (Sweden)

    Pasupathi Sundaramoorthy

    Full Text Available Cancer cell motility is a key phenomenon regulating invasion and metastasis. Focal adhesion kinase (FAK plays a major role in cellular adhesion and metastasis of various cancers. The relationship between dietary supplementation of calcium and colon cancer has been extensively investigated. However, the effect of calcium (Ca2+ supplementation on calpain-FAK-motility is not clearly understood. We sought to identify the mechanism of FAK cleavage through Ca2+ bound lactate (CaLa, its downstream signaling and role in the motility of human colon cancer cells. We found that treating HCT116 and HT-29 cells with CaLa immediately increased the intracellular Ca2+ (iCa2+ levels for a prolonged period of time. Ca2+ influx induced cleavage of FAK into an N-terminal FAK (FERM domain in a dose-dependent manner. Phosphorylated FAK (p-FAK was also cleaved in to its p-N-terminal FAK. CaLa increased colon cancer cells motility. Calpeptin, a calpain inhibitor, reversed the effects of CaLa on FAK and pFAK cleavage in both cancer cell lines. The cleaved FAK translocates into the nucleus and modulates p53 stability through MDM2-associated ubiquitination. CaLa-induced Ca2+ influx increased the motility of colon cancer cells was mediated by calpain activity through FAK and pFAK protein destabilization. In conclusion, these results suggest that careful consideration may be given in deciding dietary Ca2+ supplementation to patient undergoing treatment for metastatic cancer.

  5. Restriction fragment length polymorphism in calpain (CAPN2 gene in crossbred cattle

    Directory of Open Access Journals (Sweden)

    Maria Aparecida Cassiano Lara

    2012-12-01

    Full Text Available With advances in molecular genetics have been possible to predict the genetic value of the animal, in particular its potential to transmit desired characters to their offspring, including characters difficult to evaluate or with low heritability, as is the case of the meat tenderization. It is known that Bos taurus indicus features differences in meat tenderization, being assigned this variability to their lowest proteolysis post-mortem, as result of high activity of calpastatin. This inhibitor decreases the activity of calpain, which are the enzymes responsible for the degradation of muscle fibers during the maturation of the meat. Moreover, there were previously observed differences in the frequencies of allele A of calpain among European breeds (Hereford, Aberdeen Angus and Holstein and Bos taurus indicus (Gir, Guzerá and Nelore. This variability has been related to tenderness of meat, as cattle with Bos taurus taurus origin have more tender meat than Bos taurus indicus, showing small values of shear force. One explanation is that the Capn2A product could confer greater proteolytic activity than the encoded by the allele Capn2B. If allele A is associated with tender meat, it will be possible the early identification of the animals that have the potential to produce meat with qualities that attend the needs of the consumer market, in order to add economic value to the final product of the animal production chain. For this reason, biochemical and genetic studies related to calpain and calpastatin systems have been considered promising for the clarification of the physiological changes that occur in muscle structure during the period post-mortem, whose results have contributed to the improvement of meat quality. The objectives of this study were to investigate the RFLP in calpain (Capn2 gene and its relation with meat tenderization in 252 crossbred (Bos taurus taurus x Bos taurus indicus. The analyses were carried through by PCR-RFLP technique

  6. Expression of the gene for large subunit of m-calpain is elevated in ...

    Indian Academy of Sciences (India)

    Calpain is an intracellular nonlysosomal protease involved in essential regulatory or processing functions of the cell, mediated by physiological concentrations of Ca2З. However, in an environment of abnormal intracellular calcium, such as that seen in Duchenne muscular dystrophy (DMD), calpain is suggested to cause ...

  7. Calpain inhibition prevents amyloid-beta-induced neurodegeneration and associated behavioral dysfunction in rats

    NARCIS (Netherlands)

    Granic, Ivica; Nyakas, Csaba; Luiten, Paul G. M.; Eisel, Ulrich L. M.; Halmy, Laszlo G.; Gross, Gerhard; Schoemaker, Hans; Moeller, Achim; Nimmrich, Volker

    2010-01-01

    Amyloid-beta (A beta) is toxic to neurons and such toxicity is - at least in part - mediated via the NMDA receptor. Calpain, a calcium dependent cystein protease, is part of the NMDA receptor-induced neurodegeneration pathway, and we previously reported that inhibition of calpain prevents

  8. Growth and development of skeletal muscle in mu-calpain knockout mice

    Science.gov (United States)

    The calpain system has been identified as a potential candidate in muscle growth and development due to its role in a variety of cellular processes such as cytoskeletal remodeling and myogenesis. The objective of this study was to evaluate growth and development of skeletal muscle in mu-calpain kno...

  9. S-nitrosation of calpains is associated with cardioprotection in myocardial I/R injury.

    Science.gov (United States)

    Totzeck, Matthias; Korste, Sebastian; Miinalainen, Ilkka; Hendgen-Cotta, Ulrike B; Rassaf, Tienush

    2017-07-01

    Myocardial infarction remains the single leading cause of death worldwide. Upon reperfusion of occluded arteries, deleterious cellular mediators particularly located at the mitochondria level can be activated, thus limiting the outcome in patients. This may lead to the so-called ischemia/reperfusion (I/R) injury. Calpains are cysteine proteases and mediators of caspase-independent cell death. Recently, they have emerged as central transmitters of cellular injury in several cardiac pathologies e.g. hypertrophy and acute I/R injury. Here we investigated the role of cardiac calpains in acute I/R in relation to mitochondrial integrity and whether calpains can be effectively inhibited by posttranslational modification by S-nitrosation. Taking advantage of the a cardiomyocyte cell line (HL1), we determined S-nitrosation by the Biotin-switch approach, cell viability and intracellular calcium concentration after simulated ischemia and reoxygenation - all in dependence of supplementation with nitrite, which is known as an 'hypoxic nitric oxide (NO) donor'. Likewise, using an in vivo I/R model, calpain S-nitrosation, calpain activity and myocardial I/R injury were characterized in vivo. Nitrite administration resulted in an increased S-nitrosation of calpains, and this was associated with an improved cell-survival. No impact was detected on calcium levels. In line with these in vitro experiments, nitrite initiated calpain S-nitrosation in vivo and caused an infarct sparing effect in an in vivo myocardial I/R model. Using electron microscopy in combination with immuno-gold labeling we determined that calpain 10 increased, while calpain 2 decreased in the course of I/R. Nitrite, in turn, prevented an I/R induced increase of calpains 10 at mitochondria and reduced levels of calpain 1. Lethal myocardial injury remains a key aspect of myocardial I/R. We show that calpains, as key players in caspase-independent apoptosis, increasingly locate at mitochondria following I

  10. Expression of the calpain system is associated with poor clinical outcome in gastro-oesophageal adenocarcinomas.

    Science.gov (United States)

    Storr, Sarah J; Pu, Xuan; Davis, Jillian; Lobo, Dileep; Reece-Smith, Alex M; Parsons, Simon L; Madhusudan, Srinivasan; Martin, Stewart G

    2013-11-01

    Surgery is critical in the management of gastro-oesophageal cancer, and the addition of neo-adjuvant chemotherapy has proved to be of benefit. The calpain system has been implicated in tumour progression and response to various anti-cancer therapies, and therefore expression of the system was determined in this tumour type. Two cohorts of gastro-oesophageal adenocarcinomas were investigated for calpain-1, calpain-2, calpain-9 and calpastatin expression using conventional immunohistochemistry. 88 patients who received neo-adjuvant chemotherapy and 140 patients who received surgery alone were investigated using a tissue microarray approach. Calpain-1, calpain-2 and calpastatin expression was associated with adverse cancer-specific survival in the neo-adjuvant cohort (P = 0.004, P = 0.001 and P = 0.012 respectively); which remained significant in multivariate analysis (Hazard ratio (HR) = 0.337; 95% confidence interval (CI) = 0.140-0.81; P = 0.015, HR = 0.375; 95% CI = 0.165-0.858; P = 0.020 and HR = 0.481; 95% CI = 0.257-0.900; P = 0.022 respectively). Calpain-1 and calpastatin expression was also associated with adverse cancer specific survival in the primary surgery cohort (P = 0.001 and P = 0.013 respectively); which remained significant in multivariate analysis (HR = 0.309; 95% CI = 0.159-0.601; P = 0.001 and HR = 0.418; 95% CI = 0.205-0.850; P = 0.016 respectively). Calpain-9 expression was not associated with cancer-specific survival in the neo-adjuvant and primary surgery cohorts. Determining the expression levels of calpain-1, calpain-2 and calpastatin may provide clinically relevant prognostic information for gastro-oesophageal adenocarcinomas; these findings warrant further studies in larger cohorts of patients.

  11. Vital Role of the Calpain-Calpastatin System for Placental-Integrity-Dependent Embryonic Survival▿†

    Science.gov (United States)

    Takano, Jiro; Mihira, Naomi; Fujioka, Ryo; Hosoki, Emi; Chishti, Athar H.; Saido, Takaomi C.

    2011-01-01

    Although the calpain-calpastatin system has been implicated in a number of pathological conditions, its normal physiological role remains largely unknown. To investigate the functions of this system, we generated conventional and conditional calpain-2 knockout mice. The conventional calpain-2 knockout embryos died around embryonic day 15, preceded by cell death associated with caspase activation and DNA fragmentation in placental trophoblasts. In contrast, conditional knockout mice in which calpain-2 is expressed in the placenta but not in the fetus were spared. These results suggest that calpain-2 contributes to trophoblast survival via suppression of caspase activation. Double-knockout mice also deficient in calpain-1 and calpastatin resulted in accelerated and rescued embryonic lethality, respectively, suggesting that calpain-1 and -2 at least in part share similar in vivo functions under the control of calpastatin. Triple-knockout mice exhibited early embryonic lethality, a finding consistent with the notion that this protease system is vital for embryonic survival. PMID:21791606

  12. Effects of calpain genotypes on meat tenderness and carcass traits of Angus bulls.

    Science.gov (United States)

    Chung, H Y; Davis, M E

    2011-10-01

    Relationships of the calpain system with meat tenderness and carcass traits were examined for 94 purebred Angus bulls with genotypes of the calpain classified by RFLP (restriction fragment length polymorphism) and SSCP (Single strand conformation polymorphism) analysis. Designing of primers based on the calpain regulatory subunit (CAPNS) and u-calpian (CAPN1) genes. Bulls from 15 months of age were slaughtered, and carcass traits, including fat thickness (FAT); longissimus muscle area (LMA); percentage of kidney, pelvic, and heart fat (KPH); hot carcass weight (HCW); marbling score (MAR); and quality grade (QUL), were analyzed. Measurements regarding meat tenderness involved activities of calpastatin (CAC), u-calpain (UAC), m-calpain (MAC), Warner-Bratzler Shear Force (WBS) and myofibril fragmentation index (MFI). Statistical significances of the calpain genotypes accounted for variations in MAR and QUL at CAPNS locus, and both loci explained variations of UAC and MAC. Significant mean differences in genotypes of CAPNS locus were found for MAR (BB > AB > AA) and QUL (AB > BB > AA). UAC showed significant correlations with MAC, CAC, MFI, FAT, and MAR, and we found that MAC correlated with WBS, FAT, HCW, MAR, and QUL. Strong positive correlation detected between LMA and HCW, and MAR and QUL, and a negative correlation between MFI and MAR was estimated. From the result it may be possible to use the calpain genotypes classified by RFLP and SSCP analysis in marker assisted selection programs to estimate UAC and MAC precisely regardless meat tenderness and to improve MAR and QUL of beef cattle.

  13. Suppression of calpain expression by NSAIDs is associated with inhibition of cell migration in rat duodenum.

    Science.gov (United States)

    Silver, Kristopher; Littlejohn, A; Thomas, Laurel; Bawa, Bhupinder; Lillich, James D

    2017-05-15

    Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the alleviation of pain and inflammation, but these drugs are also associated with a suite of negative side effects. Gastrointestinal (GI) toxicity is particularly concerning since it affects an estimated 70% of individuals taking NSAIDs routinely, and evidence suggests the majority of toxicity is occurring in the small intestine. Traditionally, NSAID-induced GI toxicity has been associated with indiscriminate inhibition of cyclooxygenase isoforms, but other mechanisms, including inhibition of cell migration, intestinal restitution, and wound healing, are likely to contribute to toxicity. Previous efforts demonstrated that treatment of cultured intestinal epithelial cells (IEC) with NSAIDs inhibits expression and activity of calpain proteases, but the effects of specific inhibition of calpain expression in vitro or the effects of NSAIDs on intestinal cell migration in vivo remain to be determined. Accordingly, we examined the effect of suppression of calpain protease expression with siRNA on cell migration in cultured IECs and evaluated the effects of NSAID treatment on epithelial cell migration and calpain protease expression in rat duodenum. Our results show that calpain siRNA inhibits protease expression and slows migration in cultured IECs. Additionally, NSAID treatment of rats slowed migration up the villus axis and suppressed calpain expression in duodenal epithelial cells. Our results are supportive of the hypothesis that suppression of calpain expression leading to slowing of cell migration is a potential mechanism through which NSAIDs cause GI toxicity. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Selective pseudohypertrophy of vastus medialis muscles associated with calpain 3 deficiency.

    Science.gov (United States)

    Vattemi, Gaetano; Neri, Marcella; Marini, Matteo; Gualandi, Francesca; Tonin, Paola; Bertolasi, Laura; Guglielmi, Valeria; Catalli, Claudio; Novelli, Giuseppe; Ferlini, Alessandra; Tomelleri, Giuliano

    2012-09-01

    Calpain 3 deficiency causes limb girdle muscular dystrophy type 2A, which is one of the most common forms of limb girdle muscular dystrophy. Nevertheless, calpainopathy is not always associated with mutations in the specific gene and secondary reduction in protein expression has been described. We report a case of a 43-year-old man who complained of thigh muscle stiffness and had muscle hypertrophy of both vastus medialis with prolonged myotonic contraction by percussion. A muscle biopsy showed dystrophic features and calpain 3 deficiency was shown by immunoblot analysis although mutations in the specific gene were not found. Known cases of secondary calpain 3 protein deficiency were ruled out and mutations in MD1 and MD2 genes were excluded. This patient represents the first case of calpain 3 deficiency with selective pseudohypertrophy of vastus medialis muscles.

  15. Molecular determinants of survival motor neuron (SMN protein cleavage by the calcium-activated protease, calpain.

    Directory of Open Access Journals (Sweden)

    Jennifer L Fuentes

    2010-12-01

    Full Text Available Spinal muscular atrophy (SMA is a leading genetic cause of childhood mortality, caused by reduced levels of survival motor neuron (SMN protein. SMN functions as part of a large complex in the biogenesis of small nuclear ribonucleoproteins (snRNPs. It is not clear if defects in snRNP biogenesis cause SMA or if loss of some tissue-specific function causes disease. We recently demonstrated that the SMN complex localizes to the Z-discs of skeletal and cardiac muscle sarcomeres, and that SMN is a proteolytic target of calpain. Calpains are implicated in muscle and neurodegenerative disorders, although their relationship to SMA is unclear. Using mass spectrometry, we identified two adjacent calpain cleavage sites in SMN, S192 and F193. Deletion of small motifs in the region surrounding these sites inhibited cleavage. Patient-derived SMA mutations within SMN reduced calpain cleavage. SMN(D44V, reported to impair Gemin2 binding and amino-terminal SMN association, drastically inhibited cleavage, suggesting a role for these interactions in regulating calpain cleavage. Deletion of A188, a residue mutated in SMA type I (A188S, abrogated calpain cleavage, highlighting the importance of this region. Conversely, SMA mutations that interfere with self-oligomerization of SMN, Y272C and SMNΔ7, had no effect on cleavage. Removal of the recently-identified SMN degron (Δ268-294 resulted in increased calpain sensitivity, suggesting that the C-terminus of SMN is important in dictating availability of the cleavage site. Investigation into the spatial determinants of SMN cleavage revealed that endogenous calpains can cleave cytosolic, but not nuclear, SMN. Collectively, the results provide insight into a novel aspect of the post-translation regulation of SMN.

  16. Vital Role of the Calpain-Calpastatin System for Placental-Integrity-Dependent Embryonic Survival▿†

    OpenAIRE

    Takano, Jiro; Mihira, Naomi; Fujioka, Ryo; Hosoki, Emi; Chishti, Athar H.; Saido, Takaomi C.

    2011-01-01

    Although the calpain-calpastatin system has been implicated in a number of pathological conditions, its normal physiological role remains largely unknown. To investigate the functions of this system, we generated conventional and conditional calpain-2 knockout mice. The conventional calpain-2 knockout embryos died around embryonic day 15, preceded by cell death associated with caspase activation and DNA fragmentation in placental trophoblasts. In contrast, conditional knockout mice in which c...

  17. Effect of Nutrient Restriction and Re-Feeding on Calpain Family Genes in Skeletal Muscle of Channel Catfish (Ictalurus punctatus)

    OpenAIRE

    Elena Preziosa; Shikai Liu; Genciana Terova; Xiaoyu Gao; Hong Liu; Huseyin Kucuktas; Jeffery Terhune; Zhanjiang Liu

    2013-01-01

    BACKGROUND: Calpains, a superfamily of intracellular calcium-dependent cysteine proteases, are involved in the cytoskeletal remodeling and wasting of skeletal muscle. Calpains are generated as inactive proenzymes which are activated by N-terminal autolysis induced by calcium-ions. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we characterized the full-length cDNA sequences of three calpain genes, clpn1, clpn2, and clpn3 in channel catfish, and assessed the effect of nutrient restriction and ...

  18. UniProt search blastx result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 Q8R4Z1|SPA12_RAT Serpin A12 precursor (Visceral adipose-specific serpin) (Visceral adipo...se tissue-derived serine protease inhibitor) (Vaspin) - Rattus norvegicus (Rat) 5.00E-14 ...

  19. UniProt search blastx result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 Q7TMF5|SPA12_MOUSE Serpin A12 precursor (Visceral adipose-specific serpin) (Visceral adi...pose tissue-derived serine protease inhibitor) (Vaspin) - Mus musculus (Mouse) 4.00E-19 ...

  20. UniProt search blastx result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 Q8IW75|SPA12_HUMAN Serpin A12 precursor (Visceral adipose-specific serpin) (Visceral adi...pose tissue-derived serine protease inhibitor) (Vaspin) (OL-64) - Homo sapiens (Human) 3.00E-14 ...

  1. SwissProt search result: AK104199 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104199 006-304-A08 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respon...sive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  2. SwissProt search result: AK073781 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK073781 J033068H10 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respons...ive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 1e-15 ...

  3. SwissProt search result: AK069449 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK069449 J023020E21 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respons...ive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 2e-15 ...

  4. SwissProt search result: AK107547 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK107547 002-130-A05 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respon...sive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 1e-14 ...

  5. SwissProt search result: AK070043 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070043 J023039E14 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respons...ive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  6. SwissProt search result: AK104366 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104366 001-035-D07 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respon...sive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 3e-11 ...

  7. SwissProt search result: AK242300 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242300 J075192P11 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respons...ive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 9e-15 ...

  8. SwissProt search result: AK102207 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102207 J033087H03 (Q8BJZ3) Transmembrane BAX inhibitor motif containing protein 1 (RECS1 protein) (Respons...ive to centrifugal force and shear stress gene 1 protein) TMBI1_MOUSE 1e-15 ...

  9. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC063756.1 BC063756 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 3A, mRNA (cDNA clone MGC:70040 IMAGE:30288465), complete cds. ROD 1e-26 0 ...

  10. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 10, mRNA (cDNA clone MGC:116870 IMAGE:40004824), complete cds. PRI 6e-32 1 ... ...AK243629 J100087B22 BC096217.1 BC096217 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  11. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC099584.1 BC099584 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 3B, mRNA (cDNA clone MGC:118408 IMAGE:30291007), complete cds. ROD 1e-27 0 ...

  12. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC069938.1 BC069938 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 10, mRNA (cDNA clone MGC:78236 IMAGE:5035202), complete cds. ROD 1e-21 0 ...

  13. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 10, mRNA (cDNA clone MGC:116872 IMAGE:40004826), complete cds. PRI 6e-32 1 ... ...AK243629 J100087B22 BC096219.1 BC096219 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  14. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 4, mRNA (cDNA clone MGC:27150 IMAGE:4047303), complete cds. PRI 3e-32 1 ... ...AK243629 J100087B22 BC017401.1 BC017401 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  15. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 6, mRNA (cDNA clone MGC:2180 IMAGE:3051381), complete cds. PRI 1e-31 1 ... ...AK243629 J100087B22 BC001394.2 BC001394 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  16. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 1, mRNA (cDNA clone MGC:9340 IMAGE:3456154), complete cds. PRI 6e-29 1 ... ...AK243629 J100087B22 BC009015.1 BC009015 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  17. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 10, mRNA (cDNA clone MGC:78236 IMAGE:5035202), complete cds. ROD 4e-30 1 ... ...AK243629 J100087B22 BC069938.1 BC069938 Mus musculus serine (or cysteine) proteinase inhibitor, clade B (ova

  18. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC062134.1 BC062134 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 12, mRNA (cDNA clone MGC:69970 IMAGE:30287757), complete cds. ROD 2e-20 0 ...

  19. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 2, mRNA (cDNA clone MGC:13616 IMAGE:4281085), complete cds. PRI 1e-26 1 ... ...AK243629 J100087B22 BC012609.1 BC012609 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  20. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC029736.1 BC029736 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 10, mRNA (cDNA clone MGC:25706 IMAGE:3676050), complete cds. ROD 1e-21 0 ...

  1. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 6, mRNA (cDNA clone MGC:111370 IMAGE:30563105), complete cds. PRI 8e-32 1 ... ...AK243629 J100087B22 BC098564.1 BC098564 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  2. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 3, mRNA (cDNA clone MGC:12244 IMAGE:4101988), complete cds. PRI 4e-32 1 ... ...AK243629 J100087B22 BC005224.1 BC005224 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  3. GenBank blastx search result: AK287588 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287588 J065045K19 BC010313.1 BC010313 Mus musculus serine (or cysteine) peptidase inhibitor, clade B (oval...bumin), member 11, mRNA (cDNA clone MGC:6485 IMAGE:2646766), complete cds. ROD 2e-22 0 ...

  4. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 7, mRNA (cDNA clone MGC:97111 IMAGE:7262336), complete cds. PRI 1e-25 1 ... ...AK243629 J100087B22 BC069442.1 BC069442 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  5. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 3B, mRNA (cDNA clone MGC:118408 IMAGE:30291007), complete cds. ROD 2e-34 1 ... ...AK243629 J100087B22 BC099584.1 BC099584 Mus musculus serine (or cysteine) proteinase inhibitor, clade B (ova

  6. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 7, mRNA (cDNA clone MGC:97119 IMAGE:7262348), complete cds. PRI 1e-25 1 ... ...AK243629 J100087B22 BC069547.1 BC069547 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  7. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 12, mRNA (cDNA clone MGC:69970 IMAGE:30287757), complete cds. ROD 9e-26 1 ... ...AK243629 J100087B22 BC062134.1 BC062134 Mus musculus serine (or cysteine) proteinase inhibitor, clade B (ova

  8. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 11, mRNA (cDNA clone MGC:97176 IMAGE:7262420), complete cds. PRI 7e-28 1 ... ...AK243629 J100087B22 BC069596.1 BC069596 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  9. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 7, mRNA (cDNA clone MGC:97103 IMAGE:7262324), complete cds. PRI 1e-25 1 ... ...AK243629 J100087B22 BC069417.1 BC069417 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  10. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 10, mRNA (cDNA clone MGC:25706 IMAGE:3676050), complete cds. ROD 4e-30 1 ... ...AK243629 J100087B22 BC029736.1 BC029736 Mus musculus serine (or cysteine) proteinase inhibitor, clade B (ova

  11. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 10, mRNA (cDNA clone MGC:116871 IMAGE:40004825), complete cds. PRI 5e-32 1 ... ...AK243629 J100087B22 BC096218.1 BC096218 Homo sapiens serine (or cysteine) proteinase inhibitor, clade B (ova

  12. GenBank blastx search result: AK243629 [KOME

    Lifescience Database Archive (English)

    Full Text Available lbumin), member 11, mRNA (cDNA clone MGC:6485 IMAGE:2646766), complete cds. ROD 2e-28 1 ... ...AK243629 J100087B22 BC010313.1 BC010313 Mus musculus serine (or cysteine) proteinase inhibitor, clade B (ova

  13. Distinct molecular regulation of glycogen synthase kinase-3alpha isozyme controlled by its N-terminal region: functional role in calcium/calpain signaling.

    Science.gov (United States)

    Azoulay-Alfaguter, Inbar; Yaffe, Yakey; Licht-Murava, Avital; Urbanska, Malgorzata; Jaworski, Jacek; Pietrokovski, Shmuel; Hirschberg, Koret; Eldar-Finkelman, Hagit

    2011-04-15

    Glycogen synthase kinase-3 (GSK-3) is expressed as two isozymes α and β. They share high similarity in their catalytic domains but differ in their N- and C-terminal regions, with GSK-3α having an extended glycine-rich N terminus. Here, we undertook live cell imaging combined with molecular and bioinformatic studies to understand the distinct functions of the GSK-3 isozymes focusing on GSK-3α N-terminal region. We found that unlike GSK-3β, which shuttles between the nucleus and cytoplasm, GSK-3α was excluded from the nucleus. Deletion of the N-terminal region of GSK-3α resulted in nuclear localization, and treatment with leptomycin B resulted in GSK-3α accumulation in the nucleus. GSK-3α rapidly accumulated in the nucleus in response to calcium or serum deprivation, and accumulation was strongly inhibited by the calpain inhibitor calpeptin. This nuclear accumulation was not mediated by cleavage of the N-terminal region or phosphorylation of GSK-3α. Rather, we show that calcium-induced GSK-3α nuclear accumulation was governed by GSK-3α binding with as yet unknown calpain-sensitive protein or proteins; this binding was mediated by the N-terminal region. Bioinformatic and experimental analyses indicated that nuclear exclusion of GSK-3α was likely an exclusive characteristic of mammalian GSK-3α. Finally, we show that nuclear localization of GSK-3α reduced the nuclear pool of β-catenin and its target cyclin D1. Taken together, these data suggest that the N-terminal region of GSK-3α is responsible for its nuclear exclusion and that binding with a calcium/calpain-sensitive product enables GSK-3α nuclear retention. We further uncovered a novel link between calcium and nuclear GSK-3α-mediated inhibition of the canonical Wnt/β-catenin pathway.

  14. Interaction between Calpain-1 and HSP90: New Insights into the Regulation of Localization and Activity of the Protease

    Science.gov (United States)

    Averna, Monica; De Tullio, Roberta; Pedrazzi, Marco; Bavestrello, Margherita; Pellegrini, Matteo; Salamino, Franca; Pontremoli, Sandro; Melloni, Edon

    2015-01-01

    Here we demonstrate that heat shock protein 90 (HSP90) interacts with calpain-1, but not with calpain-2, and forms a discrete complex in which the protease maintains its catalytic activity, although with a lower affinity for Ca2+. Equilibrium gel distribution experiments show that this complex is composed by an equal number of molecules of each protein partner. Moreover, in resting cells, cytosolic calpain-1 is completely associated with HSP90. Since calpain-1, in association with HSP90, retains its proteolytic activity, and the chaperone is displaced by calpastatin also in the absence of Ca2+, the catalytic cleft of the protease is not involved in this association. Thus, calpain-1 can form two distinct complexes depending on the availability of calpastatin in the cytosol. The occurrence of a complex between HSP90 and calpain-1, in which the protease is still activable, can prevent the complete inhibition of the protease even in the presence of high calpastatin levels. We also demonstrate that in basal cell conditions HSP90 and calpain-1, but not calpain-2, are inserted in the multi-protein N-Methyl-D-Aspartate receptor (NMDAR) complex. The amount of calpain-1 at the NMDAR cluster is not modified in conditions of increased [Ca2+]i, and this resident protease is involved in the processing of NMDAR components. Finally, the amount of calpain-1 associated with NMDAR cluster is independent from Ca2+-mediated translocation. Our findings show that HSP90 plays an important role in maintaining a given and proper amount of calpain-1 at the functional sites. PMID:25575026

  15. Influence of early pH decline on calpain activity in porcine muscle

    DEFF Research Database (Denmark)

    Pomponio, Luigi; Ertbjerg, Per; Karlsson, Anders H

    2010-01-01

    This study investigated the influence of post-mortem pH decline on calpain activity and myofibrillar degradation.From 80 pigs, 30 Longissimus dorsi (LD) muscles were selected on the basis of pH values at 3 h post-mortem and classified into groups of 10 as fast, intermediate and slow pH decline....... The rate of pH decline early post-mortem differed between the three groups, but the ultimate pH values were similar at 24 h. Calpain activity and autolysis from 1 to 72 h post-mortem were determined using casein zymography and studied in relation to myofibrillar fragmentation. Colour and drip loss were...... measured. A faster decrease in pH resulted in reduced level of l-calpain activity and increased autolysis of the enzyme, and hence an earlier loss of activity due to activation of l-calpain in muscles with a fast pH decline. Paralleling the l-calpain activation in muscles with a fast pH decline a higher...

  16. PARP1-mediated necrosis is dependent on parallel JNK and Ca2+/calpain pathways

    Science.gov (United States)

    Douglas, Diana L.; Baines, Christopher P.

    2014-01-01

    ABSTRACT Poly(ADP-ribose) polymerase-1 (PARP1) is a nuclear enzyme that can trigger caspase-independent necrosis. Two main mechanisms for this have been proposed: one involving RIP1 and JNK kinases and mitochondrial permeability transition (MPT), the other involving calpain-mediated activation of Bax and mitochondrial release of apoptosis-inducing factor (AIF). However, whether these two mechanisms represent distinct pathways for PARP1-induced necrosis, or whether they are simply different components of the same pathway has yet to be tested. Mouse embryonic fibroblasts (MEFs) were treated with either N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) or β-Lapachone, resulting in PARP1-dependent necrosis. This was associated with increases in calpain activity, JNK activation and AIF translocation. JNK inhibition significantly reduced MNNG- and β-Lapachone-induced JNK activation, AIF translocation, and necrosis, but not calpain activation. In contrast, inhibition of calpain either by Ca2+ chelation or knockdown attenuated necrosis, but did not affect JNK activation or AIF translocation. To our surprise, genetic and/or pharmacological inhibition of RIP1, AIF, Bax and the MPT pore failed to abrogate MNNG- and β-Lapachone-induced necrosis. In conclusion, although JNK and calpain both contribute to PARP1-induced necrosis, they do so via parallel mechanisms. PMID:25052090

  17. Tissue-Specific Expression of the Chicken Calpain2 Gene

    Directory of Open Access Journals (Sweden)

    Zeng-Rong Zhang

    2010-01-01

    Full Text Available We quantified chicken calpain 2 (CAPN2 expression in two Chinese chicken breeds (mountainous black-bone chicken breed [MB] and a commercial meat type chicken breed [S01] to discern the tissue and ontogenic expression pattern and its effect on muscle metabolism. Real-time quantitative PCR assay was developed for accurate measurement of the CAPN2 mRNA expression in various tissues from chickens of different ages (0, 2, 4, 6, 8, 10, and 12 weeks. Results showed that the breast muscle and leg muscle tissues had the highest expression of CAPN2 compared to the other tissues from the same individual (P<.05. Overall, the CAPN2 mRNA level exhibited a “rise” developmental change in all tissues. The S01 chicken had a higher expression of the CAPN2 mRNA in all tissues than the MB chicken. Our results suggest that chicken CAPN2 expression may be related to chicken breeds and tissues.

  18. Calpain inhibition reduces amplitude and accelerates decay of the late sodium current in ventricular myocytes from dogs with chronic heart failure.

    Directory of Open Access Journals (Sweden)

    Albertas Undrovinas

    Full Text Available Calpain is an intracellular Ca²⁺-activated protease that is involved in numerous Ca²⁺ dependent regulation of protein function in many cell types. This paper tests a hypothesis that calpains are involved in Ca²⁺-dependent increase of the late sodium current (INaL in failing heart. Chronic heart failure (HF was induced in 2 dogs by multiple coronary artery embolization. Using a conventional patch-clamp technique, the whole-cell INaL was recorded in enzymatically isolated ventricular cardiomyocytes (VCMs in which INaL was activated by the presence of a higher (1 μM intracellular [Ca²⁺] in the patch pipette. Cell suspensions were exposed to a cell- permeant calpain inhibitor MDL-28170 for 1-2 h before INaL recordings. The numerical excitation-contraction coupling (ECC model was used to evaluate electrophysiological effects of calpain inhibition in silico. MDL caused acceleration of INaL decay evaluated by the two-exponential fit (τ₁ = 42±3.0 ms τ₂ = 435±27 ms, n = 6, in MDL vs. τ₁ = 52±2.1 ms τ₂ = 605±26 control no vehicle, n = 11, and vs. τ₁ = 52±2.8 ms τ₂ = 583±37 ms n = 7, control with vehicle, P<0.05 ANOVA. MDL significantly reduced INaL density recorded at -30 mV (0.488±0.03, n = 12, in control no vehicle, 0.4502±0.0210, n = 9 in vehicle vs. 0.166±0.05pA/pF, n = 5, in MDL. Our measurements of current-voltage relationships demonstrated that the INaL density was decreased by MDL in a wide range of potentials, including that for the action potential plateau. At the same time the membrane potential dependency of the steady-state activation and inactivation remained unchanged in the MDL-treated VCMs. Our ECC model predicted that calpain inhibition greatly improves myocyte function by reducing the action potential duration and intracellular diastolic Ca²⁺ accumulation in the pulse train.Calpain inhibition reverses INaL changes in failing dog ventricular

  19. Calpains: general characteristics and role in various states of the organism

    Directory of Open Access Journals (Sweden)

    N. F. Starodub

    2014-02-01

    Full Text Available Calpains are a family of cytoplasmic calcium-dependent proteinases with papain-like activity. They participate in a variety of processes in the body: age changes, functioning of endothelium and pulmonary system, regulation of apoptosis and necrosis, development of various hypometabolic states, arterial hypertension, diabetes and chronic kidney disease, tumor growth. It is concluded that calpains, causing limited proteolysis of substrates, play an important role in a wide range of biological phenome­na. Their activity is associated with the response to the calcium-dependent signaling and the effects of aging. Inhibition of calpains activity contributes to inhibition of endothelial dysfunction, cardiovascular disease, formation of structural and functional changes in the kidney tissue, has neuroprotective effect, preventing sarcopenia, reduces inflammatory reactions caused by hyperventilation of the lungs.

  20. Tear me down: Role of calpain in the development of cardiac ventricular hypertrophy

    Science.gov (United States)

    Patterson, Cam; Portbury, Andrea; Schisler, Jonathan C.; Willis, Monte S.

    2011-01-01

    Cardiac hypertrophy develops most commonly in response to hypertension and is an independent risk factor for the development of heart failure. The mechanisms by which cardiac hypertrophy may be reversed to reduce this risk have not been fully determined to the point where mechanism-specific therapies have been developed. Recently, proteases in the calpain family have been implicated in regulating the development of cardiac hypertrophy in preclinical animal models. In this review, we summarize the molecular mechanisms by which calpain inhibition has been shown to modulate the development of cardiac (specifically ventricular) hypertrophy. The context within which calpain inhibition might be developed for therapeutic intervention of cardiac hypertrophy is then discussed. PMID:21817165

  1. Neurotropin®inhibits calpain activity upregulated by specific alternation of rhythm in temperature in the mesencephalon of rats.

    Science.gov (United States)

    Fujisawa, Hiroki; Numazawa, Takumi; Kawamura, Minoru; Naiki, Mitsuru

    2017-02-15

    Neurotropin® (NTP), an analgesic for chronic pain, has antihyperalgesic effects in specific alternation of rhythm in temperature (SART)-stressed rats. Previous studies have shown that SART stress induces hyperalgesia, as well as post-translational modification of proteins (including substrates for calpain, a calcium-dependent cysteine protease) in the mesencephalon of rats. To better understand the mechanism of action of NTP, we investigated whether SART stress activates calpain in the mesencephalon of rats and whether NTP inhibits this activation. Wistar rats were exposed to SART stress for 5days. NTP (200NU/kg/day) was administered intraperitoneally every day from the onset of SART stress. The mechanical pain threshold was measured using the Randall-Selitto test on the 6th day. Thereafter, the rat mesencephalon was immediately collected and calpain activity was examined using western blot analysis with a calpain cleavage site-specific antibody. SART stress induced hyperalgesia and increased the calpain activity in the mesencephalon of rats. In contrast, NTP treatment attenuated the hyperalgesia and prevented the increase in calpain activity in the mesencephalon of SART-stressed rats. Interestingly, a negative correlation was identified between calpain activity and mechanical pain threshold in SART-stressed rats treated with or without NTP. Furthermore, NTP inhibited calpain activity on mammalian uncoordinated-18 in rat mesencephalon homogenate and Ac-LLY-AFC as substrates in an in vitro cell-free system. Our data demonstrate that NTP treatment prevents SART stress-induced calpain activation in the mesencephalon of rats and suggests that NTP-mediated antihyperalgesia is associated with an inhibition of calpain activity in the mesencephalon. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Targeted Gene Inactivation of Calpain-1 Suppresses Cortical Degeneration Due to Traumatic Brain Injury and Neuronal Apoptosis Induced by Oxidative Stress*

    Science.gov (United States)

    Yamada, Kaori H.; Kozlowski, Dorothy A.; Seidl, Stacey E.; Lance, Steven; Wieschhaus, Adam J.; Sundivakkam, Premanand; Tiruppathi, Chinnaswamy; Chishti, Imran; Herman, Ira M.; Kuchay, Shafi M.; Chishti, Athar H.

    2012-01-01

    Calpains are calcium-regulated cysteine proteases that have been implicated in the regulation of cell death pathways. Here, we used our calpain-1 null mouse model to evaluate the function of calpain-1 in neural degeneration following a rodent model of traumatic brain injury. In vivo, calpain-1 null mice show significantly less neural degeneration and apoptosis and a smaller contusion 3 days post-injury than wild type littermates. Protection from traumatic brain injury corroborated with the resistance of calpain-1 neurons to apoptosis induced by oxidative stress. Biochemical analysis revealed that caspase-3 activation, extracellular calcium entry, mitochondrial membrane permeability, and release of apoptosis-inducing factor from mitochondria are partially blocked in the calpain-1 null neurons. These findings suggest that the calpain-1 knock-out mice may serve as a useful model system for neuronal protection and apoptosis in traumatic brain injury and other neurodegenerative disorders in which oxidative stress plays a role. PMID:22367208

  3. Inhibition of Starvation-Triggered Endoplasmic Reticulum Stress, Autophagy, and Apoptosis in ARPE-19 Cells by Taurine through Modulating the Expression of Calpain-1 and Calpain-2

    OpenAIRE

    Yuanyuan Zhang; Shu Ren; Yuci Liu; Kun Gao; Zheng Liu; Zhou Zhang

    2017-01-01

    Age-related macular degeneration (AMD) is a complex disease with multiple initiators and pathways that converge on death for retinal pigment epithelial (RPE) cells. In this study, effects of taurine on calpains, autophagy, endoplasmic reticulum (ER) stress, and apoptosis in ARPE-19 cells (a human RPE cell line) were investigated. We first confirmed that autophagy, ER stress and apoptosis in ARPE-19 cells were induced by Earle’s balanced salt solution (EBSS) through starvation to induce RPE me...

  4. Calpain system protein expression in carcinomas of the pancreas, bile duct and ampulla.

    Science.gov (United States)

    Storr, Sarah J; Zaitoun, Abed M; Arora, Arvind; Durrant, Lindy G; Lobo, Dileep N; Madhusudan, Srinivasan; Martin, Stewart G

    2012-11-09

    Pancreatic cancer, including cancer of the ampulla of Vater and bile duct, is very aggressive and has a poor five year survival rate; improved methods of patient stratification are required. We assessed the expression of calpain-1, calpain-2 and calpastatin in two patient cohorts using immunohistochemistry on tissue microarrays. The first cohort was composed of 68 pancreatic adenocarcinomas and the second cohort was composed of 120 cancers of the bile duct and ampulla. In bile duct and ampullary carcinomas an association was observed between cytoplasmic calpastatin expression and patient age (P = 0.036), and between nuclear calpastatin expression and increased tumour stage (P = 0.026) and the presence of vascular invasion (P = 0.043). In pancreatic cancer, high calpain-2 expression was significantly associated with improved overall survival (P = 0.036), which remained significant in multivariate Cox-regression analysis (hazard ratio = 0.342; 95% confidence interva l = 0.157-0.741; P = 0.007). In cancers of the bile duct and ampulla, low cytoplasmic expression of calpastatin was significantly associated with poor overall survival (P = 0.012), which remained significant in multivariate Cox-regression analysis (hazard ratio = 0.595; 95% confidence interval = 0.365-0.968; P = 0.037). The results suggest that calpain-2 and calpastatin expression is important in pancreatic cancers, influencing disease progression. The findings of this study warrant a larger follow-up study.

  5. Calpain system protein expression in carcinomas of the pancreas, bile duct and ampulla

    Directory of Open Access Journals (Sweden)

    Storr Sarah J

    2012-11-01

    Full Text Available Abstract Background Pancreatic cancer, including cancer of the ampulla of Vater and bile duct, is very aggressive and has a poor five year survival rate; improved methods of patient stratification are required. Methods We assessed the expression of calpain-1, calpain-2 and calpastatin in two patient cohorts using immunohistochemistry on tissue microarrays. The first cohort was composed of 68 pancreatic adenocarcinomas and the second cohort was composed of 120 cancers of the bile duct and ampulla. Results In bile duct and ampullary carcinomas an association was observed between cytoplasmic calpastatin expression and patient age (P = 0.036, and between nuclear calpastatin expression and increased tumour stage (P = 0.026 and the presence of vascular invasion (P = 0.043. In pancreatic cancer, high calpain-2 expression was significantly associated with improved overall survival (P = 0.036, which remained significant in multivariate Cox-regression analysis (hazard ratio = 0.342; 95% confidence interva l = 0.157-0.741; P = 0.007. In cancers of the bile duct and ampulla, low cytoplasmic expression of calpastatin was significantly associated with poor overall survival (P = 0.012, which remained significant in multivariate Cox-regression analysis (hazard ratio = 0.595; 95% confidence interval = 0.365-0.968; P = 0.037. Conclusion The results suggest that calpain-2 and calpastatin expression is important in pancreatic cancers, influencing disease progression. The findings of this study warrant a larger follow-up study.

  6. Expression of the gene for large subunit of m-calpain is elevated in ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Genetics; Volume 79; Issue 2. Expression of the gene for large subunit of m-calpain is elevated in skeletal muscle from Duchenne muscular dystrophy patients. Tajamul Hussain Harleen Mangath C. Sundaram M. P. J. S. Anandaraj. Volume 79 Issue 2 August 2000 pp 77-80 ...

  7. Association of the calpain-10 gene with type 2 diabetes in Europeans

    DEFF Research Database (Denmark)

    Tsuchiya, Takafumi; Schwarz, Peter E H; Bosque-Plata, Laura Del

    2006-01-01

    We conducted pooled and meta-analyses of the association of the calpain-10 gene (CAPN10) polymorphisms SNP-43, Indel-19 and SNP-63 individually and as haplotypes with type 2 diabetes (T2D) in 3237 patients and 2935 controls of European ancestry. In the pooled analyses, the common SNP-43*G allele ...

  8. Extrasynaptic NMDA receptors couple preferentially to excitotoxicity via calpain-mediated cleavage of STEP.

    Science.gov (United States)

    Xu, Jian; Kurup, Pradeep; Zhang, Yongfang; Goebel-Goody, Susan M; Wu, Peter H; Hawasli, Ammar H; Baum, Matthew L; Bibb, James A; Lombroso, Paul J

    2009-07-22

    NMDA receptor (NMDAR)-mediated excitotoxicity plays an important role in several CNS disorders, including epilepsy, stroke, and ischemia. Here we demonstrate the involvement of striatal-enriched protein tyrosine phosphatase (STEP) in this critical process. STEP(61) is an alternatively spliced member of the family that is present in postsynaptic terminals. In an apparent paradox, STEP(61) regulates extracellular signal-regulated kinase 1/2 (ERK1/2) and p38, two proteins with opposing functions; activated p38 promotes cell death, whereas activated ERK1/2 promotes cell survival. We found that synaptic stimulation of NMDARs promoted STEP(61) ubiquitination and degradation, concomitant with ERK1/2 activation. In contrast, extrasynaptic stimulation of NMDARs invoked calpain-mediated proteolysis of STEP(61), producing the truncated cleavage product STEP(33) and activation of p38. The calpain cleavage site on STEP was mapped to the kinase interacting motif, a domain required for substrate binding. As a result, STEP(33) neither interacts with nor dephosphorylates STEP substrates. A synthetic peptide spanning the calpain cleavage site efficiently reduced STEP(61) degradation and attenuated p38 activation and cell death in slice models. Furthermore, this peptide was neuroprotective when neurons were subjected to excitotoxicity or cortical slices were exposed to ischemic conditions. These findings suggest a novel mechanism by which differential NMDAR stimulation regulates STEP(61) to promote either ERK1/2 or p38 activation and identifies calpain cleavage of STEP(61) as a valid target for the development of neuroprotective therapy.

  9. Aspirin Has Antitumor Effects via Expression of Calpain Gene in Cervical Cancer Cells

    Directory of Open Access Journals (Sweden)

    Sang Koo Lee

    2008-01-01

    Full Text Available Aspirin and other nonsteroidal anti-inflammatory drugs show efficacy in the prevention of cancers. It is known that they can inhibit cyclooxygenases, and some studies have shown that they can induce apoptosis. Our objective in this study was to investigate the mechanism by which aspirin exerts its apoptosis effects in human cervical cancer HeLa cells. The effect of aspirin on the gene expression was studied by differential mRNA display RT-PCR. Among the isolated genes, mu-type calpain gene was upregulated by aspirin treatment. To examine whether calpain mediates the antitumor effects, HeLa cells were stably transfected with the mammalian expression vector pCR3.1 containing mu-type calpain cDNA (pCRCAL/HeLa, and tumor formations were measured in nude mice. When tumor burden was measured by day 49, HeLa cells and pCR/HeLa cells (vector control produced tumors of 2126 mm3 and 1638 mm3, respectively, while pCRCAL/HeLa cells produced markedly smaller tumor of 434 mm3 in volume. The caspase-3 activity was markedly elevated in pCRCAL/HeLa cells. The increased activity levels of caspase-3 in pCRCAL/HeLa cells, in parallel with the decreased tumor formation, suggest a correlation between caspase-3 activity and calpain protein. Therefore, we conclude that aspirin-induced calpain mediates an antitumor effect via caspase-3 in cervical cancer cells.

  10. GenBank blastx search result: AK288081 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288081 J075172F18 AF453480.1 AF453480 Burkholderia cepacia flagellar hook-basal b...ody protein (fliE), flagellar MS ring protein (fliF), flagellar motor/switch protein (fliG), putative FliI inhibitor (fliH), flagel...lar ATPase (fliI), and putative flagellar chaperone (fliJ) genes, complete cds. BCT 0.0 0 ...

  11. Seasonal variation in red deer (Cervus elaphus) venison (M. longissimus dorsi) drip loss, calpain activity, colour and tenderness.

    Science.gov (United States)

    Wiklund, E; Dobbie, P; Stuart, A; Littlejohn, R P

    2010-11-01

    Sixty four young red deer (Cervus elaphus) stags (colour display life (Pcolour display life. A clear trend of increasing fluid loss during storage, calculated as amount of purge at 14 weeks of storage minus the amount of drip loss at 1 day post-slaughter, was evident, averaging 2.5% (SEM 0.17) over the four groups. The relative activities of the calpastatin-bound calpain, μ-calpain and m-calpain all exhibited a seasonal pattern although there was no evidence (P>0.05) that this affected tenderness. There was a highly significant (P<0.001) negative regression for the average over the four storage times of drip and purge on calpastatin-bound calpain activity. Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

  12. Calpain/SHP-1 interaction by honokiol dampening peritoneal dissemination of gastric cancer in nu/nu mice.

    Directory of Open Access Journals (Sweden)

    Shing Hwa Liu

    Full Text Available BACKGROUND: Honokiol, a small-molecular weight natural product, has previously been reported to activate apoptosis and inhibit gastric tumorigenesis. Whether honokiol inhibits the angiogenesis and metastasis of gastric cancer cells remains unknown. METHODOLOGY/PRINCIPAL FINDINGS: We tested the effects of honokiol on angiogenic activity and peritoneal dissemination using in vivo, ex vivo and in vitro assay systems. The signaling responses in human gastric cancer cells, human umbilical vascular endothelial cells (HUVECs, and isolated tumors were detected and analyzed. In a xenograft gastric tumor mouse model, honokiol significantly inhibited the peritoneal dissemination detected by PET/CT technique. Honokiol also effectively attenuated the angiogenesis detected by chick chorioallantoic membrane assay, mouse matrigel plug assay, rat aortic ring endothelial cell sprouting assay, and endothelial cell tube formation assay. Furthermore, honokiol effectively enhanced signal transducer and activator of transcription (STAT-3 dephosphorylation and inhibited STAT-3 DNA binding activity in human gastric cancer cells and HUVECs, which was correlated with the up-regulation of the activity and protein expression of Src homology 2 (SH2-containing tyrosine phosphatase-1 (SHP-1. Calpain-II inhibitor and siRNA transfection significantly reversed the honokiol-induced SHP-1 activity. The decreased STAT-3 phosphorylation and increased SHP-1 expression were also shown in isolated peritoneal metastatic tumors. Honokiol was also capable of inhibiting VEGF generation, which could be reversed by SHP-1 siRNA transfection. CONCLUSIONS/SIGNIFICANCE: Honokiol increases expression and activity of SPH-1 that further deactivates STAT3 pathway. These findings also suggest that honokiol is a novel and potent inhibitor of angiogenesis and peritoneal dissemination of gastric cancer cells, providing support for the application potential of honokiol in gastric cancer therapy.

  13. Downregulations of CD36 and Calpain-1, Inflammation, and Atherosclerosis by Simvastatin in Apolipoprotein E Knockout Mice.

    Science.gov (United States)

    Yin, Meihui; Liu, Qianqian; Yu, Lan; Yang, Yueyan; Lu, Meili; Wang, Hongxin; Luo, Duosheng; Rong, Xianglu; Tang, Futian; Guo, Jiao

    2017-01-01

    In the previous in vitro study, we found that simvastatin decreased the protein expression of CD36, the scavenger receptor, and calpain-1, the Ca2+-sensitive cysteine protease, in oxidized low-density lipoprotein (oxLDL)-treated macrophages. In this in vivo study, we investigated whether simvastatin downregulates the expression of CD36 and calpain-1 and inhibits the inflammation and atherosclerosis in apolipoprotein E knockout (ApoE KO) mice. Twenty male 6-week-old ApoE KO mice were divided into 2 groups: the ApoE KO group and the ApoE KO + simvastatin (ApoE KO + Sim) group. Atherosclerotic lesions were evaluated and the expressions of CD68, CD36, and calpain-1 in aorta were examined. Simvastatin inhibited the atherosclerotic lesion in ApoE KO mice. In addition, simvastatin reduced the contents of oxLDL, thiobarbituric acid reactive substances, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum, decreased the mRNA and protein expressions of CD36 and reduced the mRNA expression of TNF-α and IL-6 in the aortas. Furthermore, simvastatin reduced the calpain activity and the protein expression of calpain-1 in the aorta. The results suggested that the attenuation of atherosclerotic lesions in ApoE KO mice by simvastatin might be associated with the downregulations of CD36 and calpain-1 and with inflammation. © 2017 S. Karger AG, Basel.

  14. Crystal structure of calpain-3 penta-EF-hand (PEF) domain - a homodimerized PEF family member with calcium bound at the fifth EF-hand.

    Science.gov (United States)

    Partha, Sarathy K; Ravulapalli, Ravikiran; Allingham, John S; Campbell, Robert L; Davies, Peter L

    2014-07-01

    Calpains are Ca(2+) dependent intracellular cysteine proteases that cleave a wide range of protein substrates to help implement Ca(2+) signaling in the cell. The major isoforms of this enzyme family, calpain-1 and calpain-2, are heterodimers of a large and a small subunit, with the main dimer interface being formed through their C-terminal penta-EF hand (PEF) domains. Calpain-3, or p94, is a skeletal muscle-specific isoform that is genetically linked to limb-girdle muscular dystrophy. Biophysical and modeling studies with the PEF domain of calpain-3 support the suggestion that full-length calpain-3 exists as a homodimer. Here, we report the crystallization of calpain-3's PEF domain and its crystal structure in the presence of Ca(2+) , which provides evidence for the homodimer architecture of calpain-3 and supports the molecular model that places a protease core at either end of the elongated dimer. Unlike other calpain PEF domain structures, the calpain-3 PEF domain contains a Ca(2+) bound at the EF5-hand used for homodimer association. Three of the four Ca(2+) -binding EF-hands of the PEF domains are concentrated near the protease core, and have the potential to radically change the local charge within the dimer during Ca(2+) signaling. Examination of the homodimer interface shows that there would be steric clashes if the calpain-3 large subunit were to try to pair with a calpain small subunit. Database Structural data are available in the Protein Data Bank database under accession number 4OKH. © 2014 FEBS.

  15. Crystal structure of calpain-3 penta-EF-hand (PEF) domain - a homodimerized PEF family member with calcium bound at the fifth EF-hand

    Energy Technology Data Exchange (ETDEWEB)

    Partha, Sarathy K.; Ravulapalli, Ravikiran; Allingham, John S.; Campbell, Robert L.; Davies, Peter L. [Queens

    2014-08-21

    Calpains are Ca2+dependent intracellular cysteine proteases that cleave a wide range of protein substrates to help implement Ca2+ signaling in the cell. The major isoforms of this enzyme family, calpain-1 and calpain-2, are heterodimers of a large and a small subunit, with the main dimer interface being formed through their C-terminal penta-EF hand (PEF) domains. Calpain-3, or p94, is a skeletal muscle-specific isoform that is genetically linked to limb-girdle muscular dystrophy. Biophysical and modeling studies with the PEF domain of calpain-3 support the suggestion that full-length calpain-3 exists as a homodimer. Here, we report the crystallization of calpain-3's PEF domain and its crystal structure in the presence of Ca2+, which provides evidence for the homodimer architecture of calpain-3 and supports the molecular model that places a protease core at either end of the elongated dimer. Unlike other calpain PEF domain structures, the calpain-3 PEF domain contains a Ca2+ bound at the EF5-hand used for homodimer association. Three of the four Ca2+-binding EF-hands of the PEF domains are concentrated near the protease core, and have the potential to radically change the local charge within the dimer during Ca2+ signaling. Examination of the homodimer interface shows that there would be steric clashes if the calpain-3 large subunit were to try to pair with a calpain small subunit.

  16. Effects of Calcium Ion, Calpains, and Calcium Channel Blockers on Retinitis Pigmentosa

    Directory of Open Access Journals (Sweden)

    Mitsuru Nakazawa

    2011-01-01

    Full Text Available Recent advances in molecular genetic studies have revealed many of the causative genes of retinitis pigmentosa (RP. These achievements have provided clues to the mechanisms of photoreceptor degeneration in RP. Apoptosis is known to be a final common pathway in RP and, therefore, a possible therapeutic target for photoreceptor rescue. However, apoptosis is not a single molecular cascade, but consists of many different reactions such as caspase-dependent and caspase-independent pathways commonly leading to DNA fractionation and cell death. The intracellular concentration of calcium ions is also known to increase in apoptosis. These findings suggest that calpains, one of the calcium-dependent proteinases, play some roles in the process of photoreceptor apoptosis and that calcium channel antagonists may potentially inhibit photoreceptor apoptosis. Herein, the effects of calpains and calcium channel antagonists on photoreceptor degeneration are reviewed.

  17. Hypoxia reduces mature hERG channels through calpain up-regulation.

    Science.gov (United States)

    Lamothe, Shawn M; Song, WonJu; Guo, Jun; Li, Wentao; Yang, Tonghua; Baranchuk, Adrian; Graham, Charles H; Zhang, Shetuan

    2017-11-01

    Human ether-a-go-go-related gene (hERG) encodes the pore-forming subunit of the rapidly activating delayed rectifier potassium current (IKr) potassium channel, which is important for cardiac repolarization. Impairment of hERG function is the primary cause of acquired long QT syndrome, which predisposes individuals to cardiac arrhythmias and sudden death. Patients with hypoxia due to conditions such as cardiac ischemia or obstructive sleep apnea display increased incidence of cardiac arrhythmias and sudden death. We sought to understand the mechanisms that underlie hypoxia-associated cardiac arrhythmias. Using cell biology and electrophysiologic techniques, we found that hypoxic culture of hERG-expressing human embryonic kidney (HEK) cells and neonatal rat cardiomyocytes reduced hERG current/IKr and mature ERG channel expression with a concomitant increase in calpain expression. Calpain was actively released into the extracellular milieu and degraded cell-surface hERG. In contrast to hERG, the ether-a-go-go (EAG) channel was not reduced by hypoxic culture. By making chimeric channels between hERG and EAG, we identified that hypoxia-induced calpain degraded hERG by targeting its extracellular S5-pore linker. The scorpion toxin BeKm-1, which is known to selectively bind to the S5-pore linker of hERG, prevented hypoxia-induced hERG reduction. Our data provide novel information about hypoxia-mediated hERG dysfunction and may have biological and clinical implications in hypoxia-associated diseases.-Lamothe, S. M., Song, W., Guo, J., Li, W., Yang, T., Baranchuk, A., Graham, C. H., Zhang, S. Hypoxia reduces mature hERG channels through calpain up-regulation. © FASEB.

  18. Altered ubiquitin causes perturbed calcium homeostasis, hyperactivation of calpain, dysregulated differentiation, and cataract.

    Science.gov (United States)

    Liu, Ke; Lyu, Lei; Chin, David; Gao, Junyuan; Sun, Xiurong; Shang, Fu; Caceres, Andrea; Chang, Min-Lee; Rowan, Sheldon; Peng, Junmin; Mathias, Richard; Kasahara, Hideko; Jiang, Shuhong; Taylor, Allen

    2015-01-27

    Although the ocular lens shares many features with other tissues, it is unique in that it retains its cells throughout life, making it ideal for studies of differentiation/development. Precipitation of proteins results in lens opacification, or cataract, the major blinding disease. Lysines on ubiquitin (Ub) determine fates of Ub-protein substrates. Information regarding ubiquitin proteasome systems (UPSs), specifically of K6 in ubiquitin, is undeveloped. We expressed in the lens a mutant Ub containing a K6W substitution (K6W-Ub). Protein profiles of lenses that express wild-type ubiquitin (WT-Ub) or K6W-Ub differ by only ∼2%. Despite these quantitatively minor differences, in K6W-Ub lenses and multiple model systems we observed a fourfold Ca(2+) elevation and hyperactivation of calpain in the core of the lens, as well as calpain-associated fragmentation of critical lens proteins including Filensin, Fodrin, Vimentin, β-Crystallin, Caprin family member 2, and tudor domain containing 7. Truncations can be cataractogenic. Additionally, we observed accumulation of gap junction Connexin43, and diminished Connexin46 levels in vivo and in vitro. These findings suggest that mutation of Ub K6 alters UPS function, perturbs gap junction function, resulting in Ca(2+) elevation, hyperactivation of calpain, and associated cleavage of substrates, culminating in developmental defects and a cataractous lens. The data show previously unidentified connections between UPS and calpain-based degradative systems and advance our understanding of roles for Ub K6 in eye development. They also inform about new approaches to delay cataract and other protein precipitation diseases.

  19. Carbamazepine suppresses calpain-mediated autophagy impairment after ischemia/reperfusion in mouse livers

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Jae-Sung, E-mail: Jae.Kim@surgery.ufl.edu; Wang, Jin-Hee, E-mail: jin-hee.wang@surgery.ufl.edu; Biel, Thomas G., E-mail: Thomas.Biel@surgery.ufl.edu; Kim, Do-Sung, E-mail: do-sung.kim@surgery.med.ufl.edu; Flores-Toro, Joseph A., E-mail: Joseph.Flores-Toro@surgery.ufl.edu; Vijayvargiya, Richa, E-mail: rvijayvargiya@ufl.edu; Zendejas, Ivan, E-mail: ivan.zendejas@surgery.ufl.edu; Behrns, Kevin E., E-mail: Kevin.Behrns@surgery.ufl.edu

    2013-12-15

    Onset of the mitochondrial permeability transition (MPT) plays a causative role in ischemia/reperfusion (I/R) injury. Current therapeutic strategies for reducing reperfusion injury remain disappointing. Autophagy is a lysosome-mediated, catabolic process that timely eliminates abnormal or damaged cellular constituents and organelles such as dysfunctional mitochondria. I/R induces calcium overloading and calpain activation, leading to degradation of key autophagy-related proteins (Atg). Carbamazepine (CBZ), an FDA-approved anticonvulsant drug, has recently been reported to increase autophagy. We investigated the effects of CBZ on hepatic I/R injury. Hepatocytes and livers from male C57BL/6 mice were subjected to simulated in vitro, as well as in vivo I/R, respectively. Cell death, intracellular calcium, calpain activity, changes in autophagy-related proteins (Atg), autophagic flux, MPT and mitochondrial membrane potential after I/R were analyzed in the presence and absence of 20 μM CBZ. CBZ significantly increased hepatocyte viability after reperfusion. Confocal microscopy revealed that CBZ prevented calcium overloading, the onset of the MPT and mitochondrial depolarization. Immunoblotting and fluorometric analysis showed that CBZ blocked calpain activation, depletion of Atg7 and Beclin-1 and loss of autophagic flux after reperfusion. Intravital multiphoton imaging of anesthetized mice demonstrated that CBZ substantially reversed autophagic defects and mitochondrial dysfunction after I/R in vivo. In conclusion, CBZ prevents calcium overloading and calpain activation, which, in turn, suppresses Atg7 and Beclin-1 depletion, defective autophagy, onset of the MPT and cell death after I/R. - Highlights: • A mechanism of carbamazepine (CBZ)-induced cytoprotection in livers is proposed. • Impaired autophagy is a key event contributing to lethal reperfusion injury. • The importance of autophagy is extended and confirmed in an in vivo model. • CBZ is a potential

  20. Blueberry polyphenols prevent cardiomyocyte death by preventing calpain activation and oxidative stress.

    Science.gov (United States)

    Louis, Xavier Lieben; Thandapilly, Sijo Joseph; Kalt, Wilhelmina; Vinqvist-Tymchuk, Melinda; Aloud, Basma Milad; Raj, Pema; Yu, Liping; Le, Hoa; Netticadan, Thomas

    2014-08-01

    The purpose of this study was to examine the efficacy of an aqueous wild blueberry extract and five wild blueberry polyphenol fractions on an in vitro model of heart disease. Adult rat cardiomyocytes were pretreated with extract and fractions, and then exposed to norepinephrine (NE). Cardiomyocyte hypertrophy, cell death, oxidative stress, apoptosis and cardiomyocyte contractile function as well as the activities of calpain, superoxide dismutase (SOD) and catalase (CAT) were measured in cardiomyocytes treated with and without NE and blueberry fraction (BF). Four of five blueberry fractions prevented cell death and cardiomyocyte hypertrophy induced by NE. Total phenolic fraction was used for all further analysis. The NE-induced increase in oxidative stress, nuclear condensation, calpain activity and lowering of SOD and CAT activities were prevented upon pretreatment with BF. Reduced contractile function was also significantly improved with BF pretreatment. Blueberry polyphenols prevent NE-induced adult cardiomyocyte hypertrophy and cell death. The protective effects of BF may be in part attributed to a reduction in calpain activity and oxidative stress.

  1. Investigation of biochemical changes of the ovine calpain 3 exon-10 polymorphism.

    Science.gov (United States)

    Muto, Yukiyo; Morton, Jim; Palmer, David

    2015-12-01

    Calpain 3 (CAPN3) is a tissue specific calpain, and its mRNA is the most expressed calpain isoform in skeletal muscles. Many mutations and polymorphisms within the human CAPN3 gene have been reported and related to limb-girdle muscular dystrophy. Several reports link CAPN3 polymorphisms and meat quality. An association between three allele variants in exon-10 of ovine CAPN3 and the yield of fat trimmed meat cuts has been reported. This research investigated the biochemical significance of polymorphic variation in CAPN3. CAPN3 mRNA sequences were obtained from muscle samples collected from lambs which were homozygous for each of the three alleles. Four single base substitutions were found besides those in exon-10, but none of them, including the variations within exon-10, caused a change in amino acid sequence. The expression of CAPN3 mRNA and the amounts of CAPN3 protein were also compared among genotypes, and no significant differences were found. These results suggest that the reported association of specific allele variants within CAPN3 exon-10 to phenotype variations were not direct effects of CAPN3 polymorphisms. Interspecies analyses of the CAPN3 sequences indicated that the sequence reported here is more likely to be the correct common ovine CAPN3 sequence than the reference sequence. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Neuroprotective Effects of Oxytocin Hormone after an Experimental Stroke Model and the Possible Role of Calpain-1.

    Science.gov (United States)

    Etehadi Moghadam, Sepideh; Azami Tameh, Abolfazl; Vahidinia, Zeinab; Atlasi, Mohammad Ali; Hassani Bafrani, Hassan; Naderian, Homayoun

    2018-03-01

    Different mechanisms will be activated during ischemic stroke. Calpain proteases play a pivotal role in neuronal death after ischemia damage through apoptosis. Anti-apoptotic activities of the oxytocin (OT) in different ischemic tissues were reported in previous studies. Recently, a limited number of studies have noted the protective effects of OT in the brain. In the present study, the neuroprotective potential of OT in an animal model of transient middle cerebral artery occlusion (tMCAO) and the possible role of calpain-1 in the penumbra region were assessed. Adult male Wistar rats underwent 1 hour of tMCAO and were treated with nasal administration of OT. After 24 hours of reperfusion, infarct size was evaluated by triphenyltetrazolium chloride. Immunohistochemical staining and Western blotting were used to examine the expression of calpain-1. Nissl staining was performed for brain tissue morphology evaluation. OT reduced the infarct volume of the cerebral cortex and striatum compared with the ischemia control group significantly (P < .05). Calpain-1 overexpression, which was caused by ischemia, decreased after OT administration (P < .05). The number of pyknotic nuclei in neurons increased dramatically in the ischemic area and OT attenuated the apoptosis of neurons in the penumbra region (P < .01). We provided evidence for the neuroprotective role of OT after tMCAO through calpain-1 attenuation. Copyright © 2018 National Stroke Association. Published by Elsevier Inc. All rights reserved.

  3. Calpain-1 Expression in Triple-Negative Breast Cancer: A Potential Prognostic Factor Independent of the Proliferative/Apoptotic Index

    Directory of Open Access Journals (Sweden)

    Shadia M. Al-Bahlani

    2017-01-01

    Full Text Available Triple-negative breast cancer (TNBC is an aggressive type of breast cancer in which calpain system plays an important role in its cellular processes including apoptosis and proliferation. Although such roles have been assessed in tumor pathogenesis, the correlation of its expression to the proliferating/apoptotic index has not been studied yet. Immunohistochemical staining of calpain-1 was performed on paraffin-embedded tissues to correlate its expression with clinicopathological variables and outcome. The proliferation activity was determined by calculating the percentage of cells expressing the Ki-67 antigen. The apoptotic index was assessed morphologically and biochemically using Haematoxylin & Eosin method and Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay, respectively. Calpain-1 was significantly expressed in TNBC tissues varying from low to high with a significant correlation to lymph node status but not with the other clinicopathological variables, suggesting its role as a prognostic factor. In addition, a positive correlation was found between both apoptotic counts assays (P<0.001, r=0.547 as well as with proliferation (P=0.045. Calpain-1 expression had no significant correlation with either proliferation (P=0.29 or apoptotic indices (P=0.071 and P=0.100. Determining calpain-1 expression may provide relevant prognostic value for TNBC cancer patients.

  4. Calpain-10 genetic polymorphisms and polycystic ovary syndrome risk: a meta-analysis and meta-regression.

    Science.gov (United States)

    Shen, Wenjing; Li, Tianren; Hu, Yanjie; Liu, Hongbo; Song, Min

    2013-12-01

    Recent evidences suggest that common functional polymorphisms in the promoter region of the Calpain-10 gene may have an impact on an individual's susceptibility to polycystic ovary syndrome (PCOS), but individually published results are inconclusive. Our meta-analysis is aimed to provide a more precise estimation of the relationships between Calpain-10 genetic polymorphisms and PCOS risk. An extensive literature search for relevant studies was conducted on PubMed, Embase, Web of Science, Cochrane Library, and CBM databases from inception through April 1st, 2013. This meta-analysis was performed using the STATA 12.0 software. The crude odds ratio (OR) with 95% confidence interval (CI) was calculated. Fourteen case-control studies were included with a total of 2123 PCOS patients and 3612 healthy controls. Nine common SNPs in the Calpain-10 gene were addressed. Our meta-analysis indicated that UCSNP-19, UCSNP-63 and UCSNP-45 polymorphisms in the Calpain-10 gene might be associated with increased PCOS risk. However, no statistically significant association was observed in UCSNP-43, UCSNP-22, UCSNP-43, UCSNP-45, UCSNP-56, UCSNP-58, and UCSNP-110 polymorphisms. Further subgroup analysis by ethnicity revealed that UCSNP-19, UCSNP-63 and UCSNP-45 polymorphisms might decrease the risk of PCOS among Asian populations, but not among Caucasian populations. The current meta-analysis indicates that UCSNP-19, UCSNP-63 and UCSNP-45 polymorphisms in the Calpain-10 gene may be risk factors for PCOS, especially among Asian populations. © 2013.

  5. Calpain-mediated proteolysis of polycystin-1 C-terminus induces JAK2 and ERK signal alterations

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Hyunho [Transplantation Research Institute, Seoul National University Medical Research Center, Seoul (Korea, Republic of); Department of Medicine, University of Maryland, Baltimore, MD (United States); Kang, Ah-Young [Transplantation Research Institute, Seoul National University Medical Research Center, Seoul (Korea, Republic of); Department of Medicine, Program of Immunology, Graduate School, Seoul National University, Seoul (Korea, Republic of); Ko, Ah-ra [Clinical Research Center, Samsung Biomedical Research Institute, Seoul (Korea, Republic of); Park, Hayne Cho [Transplantation Research Institute, Seoul National University Medical Research Center, Seoul (Korea, Republic of); Research Coordination Center for Rare Diseases, Seoul National University Hospital, Seoul (Korea, Republic of); So, Insuk [Department of Physiology, Seoul National University College of Medicine, Seoul (Korea, Republic of); Park, Jong Hoon [Department of Biological Science, Sookmyung Women’s University, Seoul (Korea, Republic of); Cheong, Hae Il [Research Coordination Center for Rare Diseases, Seoul National University Hospital, Seoul (Korea, Republic of); Department of Pediatrics, Seoul National University Children’s Hospital, Seoul (Korea, Republic of); Kidney Research Institute, Medical Research Center, Seoul National University College of Medicine, Seoul (Korea, Republic of); Hwang, Young-Hwan [Research Coordination Center for Rare Diseases, Seoul National University Hospital, Seoul (Korea, Republic of); Department of Internal Medicine, Eulji General Hospital, Eulji University College of Medicine, Seoul (Korea, Republic of); and others

    2014-01-01

    Autosomal dominant polycystic kidney disease (ADPKD), a hereditary renal disease caused by mutations in PKD1 (85%) or PKD2 (15%), is characterized by the development of gradually enlarging multiple renal cysts and progressive renal failure. Polycystin-1 (PC1), PKD1 gene product, is an integral membrane glycoprotein which regulates a number of different biological processes including cell proliferation, apoptosis, cell polarity, and tubulogenesis. PC1 is a target of various proteolytic cleavages and proteosomal degradations, but its role in intracellular signaling pathways remains poorly understood. Herein, we demonstrated that PC1 is a novel substrate for μ- and m-calpains, which are calcium-dependent cysteine proteases. Overexpression of PC1 altered both Janus-activated kinase 2 (JAK2) and extracellular signal-regulated kinase (ERK) signals, which were independently regulated by calpain-mediated PC1 degradation. They suggest that the PC1 function on JAK2 and ERK signaling pathways might be regulated by calpains in response to the changes in intracellular calcium concentration. - Highlights: • Polycystin-1 is a target of ubiquitin-independent degradation by calpains. • The PEST domain is required for calpain-mediated degradation of polycystin-1. • Polycystin-1 may independently regulate JAK2 and ERK signaling pathways.

  6. [Effect of artenisiae scopariae and poriae powder on calpain-2 expression in liver tissue from rats with obstructive jaundice].

    Science.gov (United States)

    Cui, Yubao

    2015-05-01

    To explore the eff ect of artenisiae scopariae and poriae powder (ASPD) on calpain-2 expression in liver tissue from rats with obstructive jaundice. The rat model of obstructive jaundice was established. SD rats was divided into the control group, the obstructive jaundice group, the obstructive jaundice model plus ASPD group, the obstructive jaundice model plus saline group. Th e serum levels of TBIL, ALT, AST and other biochemical indexes were detected. The pathological changes of liver tissue were evaluated by HE staining. The calpain-2 mRNA and protein expression in liver was measured by Real-time PCR and immunohistochemistry or Western blot, respectively. The calpain-2 mRNA and protein expression levels were significantly up-regulated in live tissues from the rats with obstructive jaundice in a time-dependent manner. The ASPD could inhibit the calpain-2 expression in rats with obstructive jaundice concomitant with the decreased liver damage and the improved liver function, suggesting that calpain-2 was involved in endoplasmic reticulum stress-mediated cellular apoptosis and the occurrence of obstructive jaundice. ASPD could be used for patients with obstructive jaundice to promote the recovery of liver function after operation and to reduce the incidence of complications, which provide a theoretical basis for the reasonable application of traditional Chinese medicine in the peroperative period.

  7. Predictions of Cleavability of Calpain Proteolysis by Quantitative Structure-Activity Relationship Analysis Using Newly Determined Cleavage Sites and Catalytic Efficiencies of an Oligopeptide Array.

    Science.gov (United States)

    Shinkai-Ouchi, Fumiko; Koyama, Suguru; Ono, Yasuko; Hata, Shoji; Ojima, Koichi; Shindo, Mayumi; duVerle, David; Ueno, Mika; Kitamura, Fujiko; Doi, Naoko; Takigawa, Ichigaku; Mamitsuka, Hiroshi; Sorimachi, Hiroyuki

    2016-04-01

    Calpains are intracellular Ca(2+)-regulated cysteine proteases that are essential for various cellular functions. Mammalian conventional calpains (calpain-1 and calpain-2) modulate the structure and function of their substrates by limited proteolysis. Thus, it is critically important to determine the site(s) in proteins at which calpains cleave. However, the calpains' substrate specificity remains unclear, because the amino acid (aa) sequences around their cleavage sites are very diverse. To clarify calpains' substrate specificities, 84 20-mer oligopeptides, corresponding to P10-P10' of reported cleavage site sequences, were proteolyzed by calpains, and the catalytic efficiencies (kcat/Km) were globally determined by LC/MS. This analysis revealed 483 cleavage site sequences, including 360 novel ones. Thekcat/Kms for 119 sites ranged from 12.5-1,710 M(-1)s(-1) Although most sites were cleaved by both calpain-1 and -2 with a similarkcat/Km, sequence comparisons revealed distinct aa preferences at P9-P7/P2/P5'. The aa compositions of the novel sites were not statistically different from those of previously reported sites as a whole, suggesting calpains have a strict implicit rule for sequence specificity, and that the limited proteolysis of intact substrates is because of substrates' higher-order structures. Cleavage position frequencies indicated that longer sequences N-terminal to the cleavage site (P-sites) were preferred for proteolysis over C-terminal (P'-sites). Quantitative structure-activity relationship (QSAR) analyses using partial least-squares regression and >1,300 aa descriptors achievedkcat/Kmprediction withr= 0.834, and binary-QSAR modeling attained an 87.5% positive prediction value for 132 reported calpain cleavage sites independent of our model construction. These results outperformed previous calpain cleavage predictors, and revealed the importance of the P2, P3', and P4' sites, and P1-P2 cooperativity. Furthermore, using our binary-QSAR model

  8. Mechanical stimulation of C2C12 cells increases m-calpain expression, focal adhesion plaque protein degradation and cell differentiation

    DEFF Research Database (Denmark)

    Grossi, Alberto; Lawson, Moira Ann

    have shown that m-calpain is necessary for myoblast fusion leading to the formation of muscle fibers and that inhibition of this enzyme restricts myotube formation. Whether there is a link between stretchor load induced signaling and m-calpain expression and activation is not known. Using a magnetic...... documented and has been shown to affect transcription of specific gene sequences, protein synthesis, the immune system and increase in Ca2+ influx. The past 10 years has seen a dramatic increase in the understanding of how proteolytic enzymes such as calpains can affect the growth of muscle. In vivo studies...... bead stimulation assay and a C2C12 mouse myoblast cell population, we have found that mechanical stimulation via laminin receptors leads to an increase in m-calpain expression, an enzyme found to be required for muscle cell fusion. After a short period of stimulation, m-calpain relocates into focal...

  9. Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation

    Directory of Open Access Journals (Sweden)

    Wang Ting

    2012-08-01

    Full Text Available Abstract Background Exposure to particulate matter (PM is a significant risk factor for increased cardiopulmonary morbidity and mortality. The mechanism of PM-mediated pathophysiology remains unknown. However, PM is proinflammatory to the endothelium and increases vascular permeability in vitro and in vivo via ROS generation. Objectives We explored the role of tight junction proteins as targets for PM-induced loss of lung endothelial cell (EC barrier integrity and enhanced cardiopulmonary dysfunction. Methods Changes in human lung EC monolayer permeability were assessed by Transendothelial Electrical Resistance (TER in response to PM challenge (collected from Ft. McHenry Tunnel, Baltimore, MD, particle size >0.1 μm. Biochemical assessment of ROS generation and Ca2+ mobilization were also measured. Results PM exposure induced tight junction protein Zona occludens-1 (ZO-1 relocation from the cell periphery, which was accompanied by significant reductions in ZO-1 protein levels but not in adherens junction proteins (VE-cadherin and β-catenin. N-acetyl-cysteine (NAC, 5 mM reduced PM-induced ROS generation in ECs, which further prevented TER decreases and atteneuated ZO-1 degradation. PM also mediated intracellular calcium mobilization via the transient receptor potential cation channel M2 (TRPM2, in a ROS-dependent manner with subsequent activation of the Ca2+-dependent protease calpain. PM-activated calpain is responsible for ZO-1 degradation and EC barrier disruption. Overexpression of ZO-1 attenuated PM-induced endothelial barrier disruption and vascular hyperpermeability in vivo and in vitro. Conclusions These results demonstrate that PM induces marked increases in vascular permeability via ROS-mediated calcium leakage via activated TRPM2, and via ZO-1 degradation by activated calpain. These findings support a novel mechanism for PM-induced lung damage and adverse cardiovascular outcomes.

  10. Particulate matter air pollution disrupts endothelial cell barrier via calpain-mediated tight junction protein degradation.

    Science.gov (United States)

    Wang, Ting; Wang, Lichun; Moreno-Vinasco, Liliana; Lang, Gabriel D; Siegler, Jessica H; Mathew, Biji; Usatyuk, Peter V; Samet, Jonathan M; Geyh, Alison S; Breysse, Patrick N; Natarajan, Viswanathan; Garcia, Joe G N

    2012-08-29

    Exposure to particulate matter (PM) is a significant risk factor for increased cardiopulmonary morbidity and mortality. The mechanism of PM-mediated pathophysiology remains unknown. However, PM is proinflammatory to the endothelium and increases vascular permeability in vitro and in vivo via ROS generation. We explored the role of tight junction proteins as targets for PM-induced loss of lung endothelial cell (EC) barrier integrity and enhanced cardiopulmonary dysfunction. Changes in human lung EC monolayer permeability were assessed by Transendothelial Electrical Resistance (TER) in response to PM challenge (collected from Ft. McHenry Tunnel, Baltimore, MD, particle size >0.1 μm). Biochemical assessment of ROS generation and Ca2+ mobilization were also measured. PM exposure induced tight junction protein Zona occludens-1 (ZO-1) relocation from the cell periphery, which was accompanied by significant reductions in ZO-1 protein levels but not in adherens junction proteins (VE-cadherin and β-catenin). N-acetyl-cysteine (NAC, 5 mM) reduced PM-induced ROS generation in ECs, which further prevented TER decreases and atteneuated ZO-1 degradation. PM also mediated intracellular calcium mobilization via the transient receptor potential cation channel M2 (TRPM2), in a ROS-dependent manner with subsequent activation of the Ca2+-dependent protease calpain. PM-activated calpain is responsible for ZO-1 degradation and EC barrier disruption. Overexpression of ZO-1 attenuated PM-induced endothelial barrier disruption and vascular hyperpermeability in vivo and in vitro. These results demonstrate that PM induces marked increases in vascular permeability via ROS-mediated calcium leakage via activated TRPM2, and via ZO-1 degradation by activated calpain. These findings support a novel mechanism for PM-induced lung damage and adverse cardiovascular outcomes.

  11. Passive stretch reduces calpain activity through nitric oxide pathway in unloaded soleus muscles.

    Science.gov (United States)

    Xu, Peng-Tao; Li, Quan; Sheng, Juan-Juan; Chang, Hui; Song, Zhen; Yu, Zhi-Bin

    2012-08-01

    Unloading in spaceflight or long-term bed rest induces to pronounced atrophy of anti-gravity skeletal muscles. Passive stretch partially resists unloading-induced atrophy of skeletal muscle, but the mechanism remains elusive. The aims of this study were to investigate the hypotheses that stretch tension might increase protein level of neuronal nitric oxide synthase (nNOS) in unloaded skeletal muscle, and then nNOS-derived NO alleviated atrophy of skeletal muscle by inhibiting calpain activity. The tail-suspended rats were used to unload rat hindlimbs for 2 weeks, at the same time, left soleus muscle was stretched by applying a plaster cast to fix the ankle at 35° dorsiflexion. Stretch partially resisted atrophy and inhibited the decreased protein level and activity of nNOS in unloaded soleus muscles. Unloading increased frequency of calcium sparks and elevated intracellular resting and caffeine-induced Ca(2+) concentration ([Ca(2+)]i) in unloaded soleus muscle fibers. Stretch reduced frequency of calcium sparks and restored intracellular resting and caffeine-induced Ca(2+) concentration to control levels in unloaded soleus muscle fibers. The increased protein level and activity of calpain as well as the higher degradation of desmin induced by unloading were inhibited by stretch in soleus muscles. In conclusion, these results suggest that stretch can preserve the stability of sarcoplasmic reticulum Ca(2+) release channels which prevents the elevated [Ca(2+)]i by means of keeping nNOS activity, and then the enhanced protein level and activity of calpain return to control levels in unloaded soleus muscles. Therefore, stretch can resist in part atrophy of unloaded soleus muscles.

  12. Inhibition of Calpain Prevents N-Methyl-D-aspartate-Induced Degeneration of the Nucleus Basalis and Associated Behavioral Dysfunction

    NARCIS (Netherlands)

    Nimmrich, Volker; Szabo, Robert; Nyakas, Csaba; Granic, Ivica; Reymann, Klaus G.; Schroeder, Ulrich H.; Gross, Gerhard; Schoemaker, Hans; Wicke, Karsten; Moeller, Achim; Luiten, Paul

    2008-01-01

    N-Methyl-D-aspartate( NMDA) receptor-mediated excitotoxicity is thought to underlie a variety of neurological disorders, and inhibition of either the NMDA receptor itself, or molecules of the intracellular cascade, may attenuate neurodegeneration in these diseases. Calpain, a calcium-dependent

  13. Product annotations: AK288848 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288848 J090075F08 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 3e-38 ...

  14. Product annotations: AK288925 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288925 J090082B10 BLASTN BD354172.1 PAT BD354172 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 2e-13 ...

  15. Product annotations: AK288372 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288372 J090026F05 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 1e-69 ...

  16. Product annotations: AK288996 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288996 J090088B05 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 3e-25 ...

  17. Product annotations: AK288871 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288871 J090078G09 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 6e-96 ...

  18. Product annotations: AK288766 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288766 J090067H02 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 7e-41 ...

  19. Product annotations: AK288897 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288897 J090080G08 BLASTN BD354173.1 PAT BD354173 A method for genotyping the peri...pheral region of plant sd-1 gene, and a method for examining semidwarfism of plants using the method. 5e-83 ...

  20. Product annotations: AK289108 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK289108 J090096M08 BLASTN AY641412.1 PLN AY641412 Hordeum vulgare subsp. vulgare retrotransposon Sandra...in (USP1) genes and Hv receptor-kinase-like pseudogene, complete cds; and retrotransposons Sandra6 and Sandra7, complete sequence. 4e-38 ...

  1. 46 CFR 7.165 - Kenai Peninsula, AK to Kodiak Island, AK.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Kenai Peninsula, AK to Kodiak Island, AK. 7.165 Section... BOUNDARY LINES Alaska § 7.165 Kenai Peninsula, AK to Kodiak Island, AK. (a) A line drawn from the southernmost extremity of Kenai Peninsula at longitude 151°44.0′ W. to East Amatuli Island Light; thence to the...

  2. The effects of different chilling methods on meat quality and calpain activity of pork muscle longissimus dorsi.

    Science.gov (United States)

    Xu, Yang; Huang, Ji-Chao; Huang, Ming; Xu, Bao-Cai; Zhou, Guang-Hong

    2012-01-01

    The objective of this study was to investigate the effects of conventional chilling (0 to 4 °C), rapid chilling (RC, -20 °C for 30 min, followed by 0 to 4 °C), and short-duration chilling (0 to 4 °C for 30 min, followed by 25 °C) on meat quality and calpain activity of pork muscle longissimus dorsi (LD). The muscle quality characteristics pH, color, cooking loss, pressing loss and tenderness, and calpain activities were measured 0-, 3-, 12-, and 24-h postmortem. Results show that the RC resulted in a faster temperature decline of the muscle, and prevented the meat pH and Commission Internationale de l'Eclairage L* value from declining during postmortem aging. RC also reduced meat cooking loss and pressing loss compared with the other two chilling methods. However, the chilling methods did not significantly affect meat shear force. During the first 24-h postmortem, there was not a noticeable change in the activity of m-calpain. But μ-calpain activity decreased regardless of chilling method. In the rapidly chilled carcasses, μ-calpain activity remained the same 3- and 12-h postmortem. However, in the short-duration chilled and conventionally chilled carcasses, the activity was visibly reduced. At 24-h postmortem, no clear zones on the gel were observed in all three treatments. Conventional and RC methods are commonly used for pork in commercial practice nowadays. Compared with conventional chilling, the effect of RC on quality parameters of pork varies. In recent years, short-duration chilling (SC) is widely used in many Chinese pig slaughtering facilities. However, few researchers have studied the effect of SD on pork quality. Therefore, the present study investigated the effect of different chilling methods on functionalities or quality of chilled pork meat. © 2011 Institute of Food Technologists®

  3. Effect of nutrient restriction and re-feeding on calpain family genes in skeletal muscle of channel catfish (Ictalurus punctatus).

    Science.gov (United States)

    Preziosa, Elena; Liu, Shikai; Terova, Genciana; Gao, Xiaoyu; Liu, Hong; Kucuktas, Huseyin; Terhune, Jeffery; Liu, Zhanjiang

    2013-01-01

    Calpains, a superfamily of intracellular calcium-dependent cysteine proteases, are involved in the cytoskeletal remodeling and wasting of skeletal muscle. Calpains are generated as inactive proenzymes which are activated by N-terminal autolysis induced by calcium-ions. In this study, we characterized the full-length cDNA sequences of three calpain genes, clpn1, clpn2, and clpn3 in channel catfish, and assessed the effect of nutrient restriction and subsequent re-feeding on the expression of these genes in skeletal muscle. The clpn1 cDNA sequence encodes a protein of 704 amino acids, Clpn2 of 696 amino acids, and Clpn3 of 741 amino acids. Phylogenetic analysis of deduced amino acid sequences indicate that catfish Clpn1 and Clpn2 share a sequence similarity of 61%; catfish Clpn1 and Clpn3 of 48%, and Clpn2 and Clpn3 of only 45%. The domain structure architectures of all three calpain genes in channel catfish are similar to those of other vertebrates, further supported by strong bootstrap values during phylogenetic analyses. Starvation of channel catfish (average weight, 15-20 g) for 35 days influenced the expression of clpn1 (2.3-fold decrease, Pcatfish skeletal muscle showed significant differences only during the starvation period, with a 1.2- and 1.4- fold increase (P<0.01) after 17 and 35 days of starvation, respectively. We have assessed that fasting and refeeding may provide a suitable experimental model to provide us insight into the role of calpains during fish muscle atrophy and how they respond to changes in nutrient supply.

  4. Synthesis and extended activity of triazole-containing macrocyclic protease inhibitors

    DEFF Research Database (Denmark)

    Pehere, A.D.; Pietsch, M.; Gütschow, M.

    2013-01-01

    Peptide-derived protease inhibitors are an important class of compounds with the potential to treat a wide range of diseases. Herein, we describe the synthesis of a series of triazole- containing macrocyclic protease inhibitors pre-organized into a b-strand conformation and an evaluation...... of their activity against a panel of proteases. Acyclic azidoalkyne-based aldehydes are also evaluated for comparison. The macrocyclic peptidomimetics showed considerable activity towards calpain II, cathepsin L and S, and the 20S proteasome chymotrypsin-like activity. Some of the first examples of highly potent...

  5. Calpain 3 Expression Pattern during Gastrocnemius Muscle Atrophy and Regeneration Following Sciatic Nerve Injury in Rats

    Directory of Open Access Journals (Sweden)

    Ronghua Wu

    2015-11-01

    Full Text Available Calpain 3 (CAPN3, also known as p94, is a skeletal muscle-specific member of the calpain family that is involved in muscular dystrophy; however, the roles of CAPN3 in muscular atrophy and regeneration are yet to be understood. In the present study, we attempted to explain the effect of CAPN3 in muscle atrophy by evaluating CAPN3 expression in rat gastrocnemius muscle following reversible sciatic nerve injury. After nerve injury, the wet weight ratio and cross sectional area (CSA of gastrocnemius muscle were decreased gradually from 1–14 days and then recovery from 14–28 days. The active form of CAPN3 (~62 kDa protein decreased slightly on day 3 and then increased from day 7 to 14 before a decrease from day 14 to 28. The result of linear correlation analysis showed that expression of the active CAPN3 protein level was negatively correlated with muscle wet weight ratio. CAPN3 knockdown by short interfering RNA (siRNA injection improved muscle recovery on days 7 and 14 after injury as compared to that observed with control siRNA treatment. Depletion of CAPN3 gene expression could promote myoblast differentiation in L6 cells. Based on these findings, we conclude that the expression pattern of the active CAPN3 protein is linked to muscle atrophy and regeneration following denervation: its upregulation during early stages may promote satellite cell renewal by inhibiting differentiation, whereas in later stages, CAPN3 expression may be downregulated to stimulate myogenic differentiation and enhance recovery. These results provide a novel mechanistic insight into the role of CAPN3 protein in muscle regeneration after peripheral nerve injury.

  6. Identification of different domains of calpain and calpastatin from chicken blood and their role in post-mortem aging of meat during holding at refrigeration temperatures.

    Science.gov (United States)

    Biswas, A K; Tandon, S; Beura, C K

    2016-06-01

    The aim of this study was to develop a simple, specific and rapid analytical method for accurate identification of calpain and calpastatin from chicken blood and muscle samples. The method is based on liquid-liquid extraction technique followed by casein Zymography detection. The target compounds were extracted from blood and meat samples by tris buffer, and purified and separated on anion exchange chromatography. It has been observed that buffer (pH 6.7) containing 50 mM tris-base appears to be excellent extractant as activity of analytes was maximum for all samples. The concentrations of μ-, m-calpain and calpastatin detected in the extracts of blood, breast and thigh samples were 0.28-0.55, 1.91-2.05 and 1.38-1.52 Unit/g, respectively. For robustness, the analytical method was applied to determine the activity of calpains (μ and m) in eighty postmortem muscle samples. It has been observed that μ-calpain activity in breast and thigh muscles declined very rapidly at 48 h and 24 h, respectively while activity of m-calpain remained stable. Shear force values were also declined with the increase of post-mortem aging showing the presence of ample tenderness of breast and thigh muscles. Finally, it is concluded that the method standardized for the detection of calpain and calpastatin has the potential to be applied to identify post-mortem aging of chicken meat samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Effect of nutrient restriction and re-feeding on calpain family genes in skeletal muscle of channel catfish (Ictalurus punctatus.

    Directory of Open Access Journals (Sweden)

    Elena Preziosa

    Full Text Available BACKGROUND: Calpains, a superfamily of intracellular calcium-dependent cysteine proteases, are involved in the cytoskeletal remodeling and wasting of skeletal muscle. Calpains are generated as inactive proenzymes which are activated by N-terminal autolysis induced by calcium-ions. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we characterized the full-length cDNA sequences of three calpain genes, clpn1, clpn2, and clpn3 in channel catfish, and assessed the effect of nutrient restriction and subsequent re-feeding on the expression of these genes in skeletal muscle. The clpn1 cDNA sequence encodes a protein of 704 amino acids, Clpn2 of 696 amino acids, and Clpn3 of 741 amino acids. Phylogenetic analysis of deduced amino acid sequences indicate that catfish Clpn1 and Clpn2 share a sequence similarity of 61%; catfish Clpn1 and Clpn3 of 48%, and Clpn2 and Clpn3 of only 45%. The domain structure architectures of all three calpain genes in channel catfish are similar to those of other vertebrates, further supported by strong bootstrap values during phylogenetic analyses. Starvation of channel catfish (average weight, 15-20 g for 35 days influenced the expression of clpn1 (2.3-fold decrease, P<0.05, clpn2 (1.3-fold increase, P<0.05, and clpn3 (13.0-fold decrease, P<0.05, whereas the subsequent refeeding did not change the expression of these genes as measured by quantitative real-time PCR analysis. Calpain catalytic activity in channel catfish skeletal muscle showed significant differences only during the starvation period, with a 1.2- and 1.4- fold increase (P<0.01 after 17 and 35 days of starvation, respectively. CONCLUSION/SIGNIFICANCE: We have assessed that fasting and refeeding may provide a suitable experimental model to provide us insight into the role of calpains during fish muscle atrophy and how they respond to changes in nutrient supply.

  8. Effects of concentric and repeated eccentric exercise on muscle damage and calpain-calpastatin gene expression in human skeletal muscle

    DEFF Research Database (Denmark)

    Vissing, K.; Overgaard, K.; Nedergaard, A.

    2008-01-01

    , and was compared to a control-group (n = 6). Muscle strength and soreness and plasma creatine kinase and myoglobin were measured before and during 7 days following exercise bouts. Muscle biopsies were collected from m. vastus lateralis of both legs prior to and at 3, 24 h and 7 days after exercise and quantified...... for muscle Ca2+-content and mRNA levels for calpain isoforms and calpastatin. Exercise reduced muscle strength and increased muscle soreness predominantly in the eccentric leg (P creatine kinase and myoglobin were all attenuated after the repeated...... not change at any specific time point post-exercise for either intervention. Our mRNA results suggest a regulation on the calpain-calpastatin expression response to muscle damaging eccentric exercise, but not concentric exercise. Although a repeated bout effect was demonstrated in terms of muscle function...

  9. Calpain and STriatal-Enriched protein tyrosine phosphatase (STEP) activation contribute to extrasynaptic NMDA receptor localization in a Huntington's disease mouse model.

    Science.gov (United States)

    Gladding, Clare M; Sepers, Marja D; Xu, Jian; Zhang, Lily Y J; Milnerwood, Austen J; Lombroso, Paul J; Raymond, Lynn A

    2012-09-01

    In Huntington's disease (HD), the mutant huntingtin (mhtt) protein is associated with striatal dysfunction and degeneration. Excitotoxicity and early synaptic defects are attributed, in part, to altered NMDA receptor (NMDAR) trafficking and function. Deleterious extrasynaptic NMDAR localization and signalling are increased early in yeast artificial chromosome mice expressing full-length mhtt with 128 polyglutamine repeats (YAC128 mice). NMDAR trafficking at the plasma membrane is regulated by dephosphorylation of the NMDAR subunit GluN2B tyrosine 1472 (Y1472) residue by STriatal-Enriched protein tyrosine Phosphatase (STEP). NMDAR function is also regulated by calpain cleavage of the GluN2B C-terminus. Activation of both STEP and calpain is calcium-dependent, and disruption of calcium homeostasis occurs early in the HD striatum. Here, we show increased calpain cleavage of GluN2B at both synaptic and extrasynaptic sites, and elevated extrasynaptic total GluN2B expression in the YAC128 striatum. Calpain inhibition significantly reduced extrasynaptic GluN2B expression in the YAC128 but not wild-type striatum. Furthermore, calpain inhibition reduced whole-cell NMDAR current and the surface/internal GluN2B ratio in co-cultured striatal neurons, without affecting synaptic GluN2B localization. Synaptic STEP activity was also significantly higher in the YAC128 striatum, correlating with decreased GluN2B Y1472 phosphorylation. A substrate-trapping STEP protein (TAT-STEP C-S) significantly increased VGLUT1-GluN2B colocalization, as well as increasing synaptic GluN2B expression and Y1472 phosphorylation. Moreover, combined calpain inhibition and STEP inactivation reduced extrasynaptic, while increasing synaptic GluN2B expression in the YAC128 striatum. These results indicate that increased STEP and calpain activation contribute to altered NMDAR localization in an HD mouse model, suggesting new therapeutic targets for HD.

  10. Calpain-5 gene variants are associated with diastolic blood pressure and cholesterol levels

    Directory of Open Access Journals (Sweden)

    Morón Francisco J

    2007-01-01

    Full Text Available Abstract Background Genes implicated in common complex disorders such as obesity, type 2 diabetes mellitus (T2DM or cardiovascular diseases are not disease specific, since clinically related disorders also share genetic components. Cysteine protease Calpain 10 (CAPN10 has been associated with T2DM, hypertension, hypercholesterolemia, increased body mass index (BMI and polycystic ovary syndrome (PCOS, a reproductive disorder of women in which isunlin resistance seems to play a pathogenic role. The calpain 5 gene (CAPN5 encodes a protein homologue of CAPN10. CAPN5 has been previously associated with PCOS by our group. In this new study, we have analysed the association of four CAPN5 gene variants(rs948976A>G, rs4945140G>A, rs2233546C>T and rs2233549G>A with several cardiovascular risk factors related to metabolic syndrome in general population. Methods Anthropometric measurements, blood pressure, insulin, glucose and lipid profiles were determined in 606 individuals randomly chosen from a cross-sectional population-based epidemiological survey in the province of Segovia in Central Spain (Castille, recruited to investigate the prevalence of anthropometric and physiological parameters related to obesity and other components of the metabolic syndrome. Genotypes at the four polymorphic loci in CAPN5 gene were detected by polymerase chain reaction (PCR. Results Genotype association analysis was significant for BMI (p ≤ 0.041, diastolic blood pressure (p = 0.015 and HDL-cholesterol levels (p = 0.025. Different CAPN5 haplotypes were also associated with diastolic blood pressure (DBP (0.0005 ≤ p ≤ 0.006 and total cholesterol levels (0.001 ≤ p ≤ 0.029. In addition, the AACA haplotype, over-represented in obese individuals, is also more frequent in individuals with metabolic syndrome defined by ATPIII criteria (p = 0.029. Conclusion As its homologue CAPN10, CAPN5 seems to influence traits related to increased risk for cardiovascular diseases. Our

  11. Preapoptotic protease calpain-2 is frequently suppressed in adult T-cell leukemia

    Science.gov (United States)

    Ishihara, Makoto; Araya, Natsumi; Sato, Tomoo; Tatsuguchi, Ayako; Saichi, Naomi; Utsunomiya, Atae; Nakamura, Yusuke; Nakagawa, Hidewaki; Yamano, Yoshihisa

    2013-01-01

    Adult T-cell leukemia (ATL) is one of the most aggressive hematologic malignancies caused by human T-lymphotropic virus type 1 (HTLV-1) infection. The prognosis of ATL is extremely poor; however, effective strategies for diagnosis and treatment have not been established. To identify novel therapeutic targets and diagnostic markers for ATL, we employed focused proteomic profiling of the CD4+CD25+CCR4+ T-cell subpopulation in which HTLV-1–infected cells were enriched. Comprehensive quantification of 14 064 peptides and subsequent 2-step statistical analysis using 29 cases (6 uninfected controls, 5 asymptomatic carriers, 9 HTLV-1–associated myelopathy/tropical spastic paraparesis patients, 9 ATL patients) identified 91 peptide determinants that statistically classified 4 clinical groups with an accuracy rate of 92.2% by cross-validation test. Among the identified 17 classifier proteins, α-II spectrin was drastically accumulated in infected T cells derived from ATL patients, whereas its digestive protease calpain-2 (CAN2) was significantly downregulated. Further cell cycle analysis and cell growth assay revealed that rescue of CAN2 activity by overexpressing constitutively active CAN2 (Δ19CAN2) could induce remarkable cell death on ATL cells accompanied by reduction of α-II spectrin. These results support that proteomic profiling of HTLV-1–infected T cells could provide potential diagnostic biomarkers and an attractive resource of therapeutic targets for ATL. PMID:23538341

  12. KERAGAMAN GEN CALPASTATIN, CALPAIN 3 DAN MYOSTATIN PADA DOMBA DI UP3 JONGGOL

    Directory of Open Access Journals (Sweden)

    Cece Sumantri

    2012-04-01

    Full Text Available The aim of this study was to identify the genetic polymorphisms of calpastatin (CAST, calpain 3 (CAPN3 and myostatin (MSTN on local sheep at Jonggol Animal Science Teaching and Research Unit (JASTRU. A total number of 294 blood samples were collected from JASTRU. The identification of polymorhism in CAST and CAPN3 genes performed by using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP while MSTN gene by using PCR-SSCP methods. The results showed that CAST|MspI, CAST|NcoI and CAPN3|MaeII loci were polymorphic, whereas The MSTN locus was monomorphic for G (1.0. The frequency of allele M (0.87 on the locus (CAST|MspI higher than the N allele (0.13. At locus CAST|NcoI, the frequency of allele M (0.96 higher than the N allele (0.04. At the CAPN3|MaeII, allele G (0.85 and allele T (0.15. Locus CAST|NcoI has higher observed heterozygosity (Ho = 0.92 compared to CAPN3|MaeII and CAST|MspI (Ho = 0.74-0.77, however has lower compared to CAPN3|MaeII and CAST|MspI in expected of heterozygosity (He = 0.08 vs 0.23-0.26 and in index fixation (Fis = -0.04 vs 0.03-0.12.

  13. Arabidopsis CDS blastp result: AK102134 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102134 J033085F12 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  14. Arabidopsis CDS blastp result: AK104609 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104609 006-309-C02 At3g62060.1 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GI:1431629 from [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-110 ...

  15. Arabidopsis CDS blastp result: AK064882 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064882 J013000K01 At3g62060.1 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GI:1431629 from [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-110 ...

  16. Arabidopsis CDS blastp result: AK064109 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK064109 002-101-G02 At3g09410.3 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GB:CAA67728 [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-126 ...

  17. Arabidopsis CDS blastp result: AK068437 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK068437 J013156H21 At3g09410.3 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GB:CAA67728 [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 3e-61 ...

  18. Arabidopsis CDS blastp result: AK066835 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066835 J013087I16 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 1e-171 ...

  19. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At5g64740.1 68418.m08141 cellulose synthase, catalytic subunit,... putative similar to gi:2827141 cellulose synthase catalytic subunit (Ath-A), Arabidopsis thaliana 0.0 ...

  20. Arabidopsis CDS blastp result: AK110467 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK110467 002-166-G08 At3g03050.1 cellulose synthase family protein (CslD3) similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-7 (gi:962

  1. Arabidopsis CDS blastp result: AK065259 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065259 J013002J18 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  2. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At5g09870.1 68418.m01141 cellulose synthase, catalytic subunit,... putative similar to gi:2827141 cellulose synthase catalytic subunit (Ath-A), Arabidopsis thaliana 8e-28 ...

  3. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At3g03050.1 68416.m00301 cellulose synthase family protein (CslD3) similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose syntha

  4. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At3g03050.1 68416.m00301 cellulose synthase family protein (CslD3) similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose syntha

  5. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At2g21770.1 68415.m02588 cellulose synthase, catalytic subunit,... putative similar to gi:2827141 cellulose synthase catalytic subunit, Arabidopsis thaliana (Ath-A) 5e-53 ...

  6. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At3g03050.1 68416.m00301 cellulose synthase family protein (CslD3) similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose syntha

  7. Arabidopsis CDS blastp result: AK102695 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102695 J033103F21 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  8. Arabidopsis CDS blastp result: AK100523 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100523 J023100P04 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  9. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At5g05170.1 68418.m00550 cellulose synthase, catalytic subunit ...(Ath-B) nearly identical to gi:2827143, cellulose synthase, catalytic subunit (Ath-B) 5e-47 ...

  10. Arabidopsis CDS blastp result: AK068407 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK068407 J013149B08 At4g01900.1 P II nitrogen sensing protein (GLB I) identical to P II nitrogen... sensing protein GLB I (GI:7268574) [Arabidopsis thaliana]; similar to nitrogen regulatory prot

  11. Arabidopsis CDS blastp result: AK099152 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099152 J023070H02 At4g01900.1 P II nitrogen sensing protein (GLB I) identical to P II nitrogen... sensing protein GLB I (GI:7268574) [Arabidopsis thaliana]; similar to nitrogen regulatory prot

  12. Arabidopsis CDS blastp result: AK120103 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120103 J013022N16 At5g57170.1 macrophage migration inhibitory factor family prote...in / MIF family protein contains Pfam profile: PF01187 Macrophage migration inhibitory factor(MIF) 3e-34 ...

  13. Arabidopsis CDS blastp result: AK240887 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240887 J065029G04 At5g01650.1 68418.m00081 macrophage migration inhibitory factor... family protein / MIF family protein contains pfam profile: PF001187 Macrophage migration inhibitory factor 3e-49 ...

  14. Arabidopsis CDS blastp result: AK121033 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121033 J023050K24 At5g01650.1 macrophage migration inhibitory factor family prote...in / MIF family protein contains pfam profile: PF001187 Macrophage migration inhibitory factor 1e-44 ...

  15. Arabidopsis CDS blastp result: AK121907 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121907 J033105N05 At5g62290.1 nucleotide-sensitive chloride conductance regulator... (ICln) family protein contains PF03517: Nucleotide-sensitive chloride conductance regulator (ICln) 2e-43 ...

  16. Arabidopsis CDS blastp result: AK070179 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070179 J023044M21 At5g62290.1 nucleotide-sensitive chloride conductance regulator... (ICln) family protein contains PF03517: Nucleotide-sensitive chloride conductance regulator (ICln) 2e-45 ...

  17. Arabidopsis CDS blastp result: AK066425 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066425 J013065E05 At1g79230.1 mercaptopyruvate sulfurtransferase (MST1) (RDH1) id...entical to mercaptopyruvate sulfurtransferase GI:6009981 and thiosulfate sulfurtransferase GI:5834508 from [Arabidopsis thaliana] 1e-122 ...

  18. Arabidopsis CDS blastp result: AK120712 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120712 J023001O05 At1g79230.1 mercaptopyruvate sulfurtransferase (MST1) (RDH1) id...entical to mercaptopyruvate sulfurtransferase GI:6009981 and thiosulfate sulfurtransferase GI:5834508 from [Arabidopsis thaliana] 1e-142 ...

  19. Arabidopsis CDS blastp result: AK288216 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288216 J090011C22 At3g09030.1 68416.m01059 potassium channel tetramerisation doma...in-containing protein contains Pfam profile PF02214: K+ channel tetramerisation domain 2e-18 ...

  20. Arabidopsis CDS blastp result: AK288216 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288216 J090011C22 At2g24240.1 68415.m02895 potassium channel tetramerisation doma...in-containing protein contains Pfam profile PF02214: K+ channel tetramerisation domain 1e-139 ...

  1. Arabidopsis CDS blastp result: AK288216 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288216 J090011C22 At4g30940.1 68417.m04393 potassium channel tetramerisation doma...in-containing protein contains Pfam profile PF02214: K+ channel tetramerisation domain 1e-139 ...

  2. Arabidopsis CDS blastp result: AK241821 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241821 J065212G04 At1g71710.1 68414.m08289 inositol polyphosphate 5-phosphatase, ...putative similar to inositol polyphosphate 5-phosphatase I [Arabidopsis thaliana] GI:10444261 2e-71 ...

  3. Arabidopsis CDS blastp result: AK241821 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241821 J065212G04 At1g71710.1 68414.m08289 inositol polyphosphate 5-phosphatase, ...putative similar to inositol polyphosphate 5-phosphatase I [Arabidopsis thaliana] GI:10444261 4e-21 ...

  4. Arabidopsis CDS blastp result: AK241408 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241408 J065159I10 At4g31900.1 68417.m04533 chromatin remodeling factor, putative ...strong similarity to chromatin remodeling factor CHD3 (PICKLE) [Arabidopsis thaliana] GI:6478518; contains P

  5. Arabidopsis CDS blastp result: AK242828 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242828 J090067I06 At4g31900.1 68417.m04533 chromatin remodeling factor, putative ...strong similarity to chromatin remodeling factor CHD3 (PICKLE) [Arabidopsis thaliana] GI:6478518; contains P

  6. Arabidopsis CDS blastp result: AK099680 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099680 J013071B03 At5g55220.1 trigger factor type chaperone family protein contai...ns Pfam profiles PF05697: Bacterial trigger factor protein (TF), PF05698: Bacterial trigger factor protein (

  7. Arabidopsis CDS blastp result: AK242550 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242550 J080319D10 At2g35630.1 68415.m04369 microtubule organization 1 protein (MO...R1) identical to microtubule organization 1 protein GI:14317953 from [Arabidopsis thaliana] 5e-44 ...

  8. Arabidopsis CDS blastp result: AK101494 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101494 J033043C06 At3g29090.1 pectinesterase family protein similar to pectineste...rase precursor GB:Q43043 [Petunia integrifolia]; contains Pfam profile: PF01095 pectinesterase 1e-130 ...

  9. Arabidopsis CDS blastp result: AK101105 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101105 J033025D11 At2g39090.1 tetratricopeptide repeat (TPR)-containing protein low similarity to prediabe...tic NOD sera-reactive autoantigen [Mus musculus] GI:6670773, anaphase-promoting com

  10. Arabidopsis CDS blastp result: AK105135 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK105135 001-102-A05 At2g27170.1 structural maintenance of chromosomes (SMC) family protein similar to basem...ent membrane-associated chondroitin proteoglycan Bamacan [Rattus norvegicus] GI:178

  11. Arabidopsis CDS blastp result: AK061773 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061773 001-039-D01 At1g50940.1 electron transfer flavoprotein alpha subunit famil...y protein contains Pfam profile: PF00766 electron transfer flavoprotein, alpha subunit 1e-105 ...

  12. Arabidopsis CDS blastp result: AK063110 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK063110 001-111-D06 At5g43430.1 electron transfer flavoprotein beta subunit family... protein contains Pfam profile: PF01012 electron transfer flavoprotein, beta subunit 1e-100 ...

  13. Arabidopsis CDS blastp result: AK105896 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK105896 001-204-F02 At1g50940.1 electron transfer flavoprotein alpha subunit famil...y protein contains Pfam profile: PF00766 electron transfer flavoprotein, alpha subunit 1e-105 ...

  14. Arabidopsis CDS blastp result: AK098986 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK098986 J013093P06 At1g50940.1 electron transfer flavoprotein alpha subunit family... protein contains Pfam profile: PF00766 electron transfer flavoprotein, alpha subunit 1e-105 ...

  15. Arabidopsis CDS blastp result: AK111285 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK111285 002-180-H08 At3g05540.1 translationally controlled tumor family protein si...milar to translationally controlled tumor protein GB:AAD10032 from [Hevea brasiliensis] 2e-28 ...

  16. Arabidopsis CDS blastp result: AK105453 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK105453 001-125-C05 At3g16640.1 translationally controlled tumor family protein si...milar to translationally controlled tumor protein GB:AAD10032 from [Hevea brasiliensis] 4e-66 ...

  17. Arabidopsis CDS blastp result: AK111243 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK111243 002-180-D03 At2g23420.1 nicotinate phosphoribosyltransferase family protei...n / NAPRTase family protein contains Pfam domain PF04095: Nicotinate phosphoribosyltransferase (NAPRTase) 2e-12 ...

  18. Arabidopsis CDS blastp result: AK067438 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK067438 J013103F09 At2g23420.1 nicotinate phosphoribosyltransferase family protein... / NAPRTase family protein contains Pfam domain PF04095: Nicotinate phosphoribosyltransferase (NAPRTase) 0.0 ...

  19. Arabidopsis CDS blastp result: AK099191 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099191 J023110O20 At2g23420.1 nicotinate phosphoribosyltransferase family protein... / NAPRTase family protein contains Pfam domain PF04095: Nicotinate phosphoribosyltransferase (NAPRTase) 0.0 ...

  20. Arabidopsis CDS blastp result: AK121140 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121140 J023077G12 At2g23420.1 nicotinate phosphoribosyltransferase family protein... / NAPRTase family protein contains Pfam domain PF04095: Nicotinate phosphoribosyltransferase (NAPRTase) 0.0 ...

  1. Arabidopsis CDS blastp result: AK119236 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK119236 001-116-D09 At2g23420.1 nicotinate phosphoribosyltransferase family protei...n / NAPRTase family protein contains Pfam domain PF04095: Nicotinate phosphoribosyltransferase (NAPRTase) 0.0 ...

  2. Arabidopsis CDS blastp result: AK060892 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060892 001-035-C02 At3g59380.1 farnesyltransferase alpha subunit, putative / FTA, putative / protein far...nesyltransferase, putative similar to farnesyltransferase alpha subunit [GI:2246442][Pisum sativum] 1e-116 ...

  3. Arabidopsis CDS blastp result: AK104483 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104483 006-301-H07 At3g59380.1 farnesyltransferase alpha subunit, putative / FTA, putative / protein far...nesyltransferase, putative similar to farnesyltransferase alpha subunit [GI:2246442][Pisum sativum] 1e-116 ...

  4. Arabidopsis CDS blastp result: AK098845 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK098845 J013000C07 At3g59380.1 farnesyltransferase alpha subunit, putative / FTA, putative / protein far...nesyltransferase, putative similar to farnesyltransferase alpha subunit [GI:2246442][Pisum sativum] 1e-116 ...

  5. Arabidopsis CDS blastp result: AK070180 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070180 J023047D02 At5g41770.1 crooked neck protein, putative / cell cycle protein..., putative similar to Swiss-Prot:P17886 crooked neck protein [Drosophila melanogaster] 1e-153 ...

  6. Arabidopsis CDS blastp result: AK103185 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103185 J033121L02 At5g41770.1 crooked neck protein, putative / cell cycle protein..., putative similar to Swiss-Prot:P17886 crooked neck protein [Drosophila melanogaster] 2e-84 ...

  7. Arabidopsis CDS blastp result: AK100478 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100478 J023095B21 At5g41770.1 crooked neck protein, putative / cell cycle protein..., putative similar to Swiss-Prot:P17886 crooked neck protein [Drosophila melanogaster] 0.0 ...

  8. Arabidopsis CDS blastp result: AK070656 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070656 J023055E24 At1g09240.1 nicotianamine synthase, putative similar to nicotian...amine synthase [Lycopersicon esculentum][GI:4753801], nicotianamine synthase 2 [Hordeum vulgare][GI:4894912] 1e-63 ...

  9. Arabidopsis CDS blastp result: AK101318 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101318 J033034D12 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multipl...ication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-125 ...

  10. Arabidopsis CDS blastp result: AK066854 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066854 J013075C10 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multipl...ication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-119 ...

  11. Arabidopsis CDS blastp result: AK104882 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104882 001-044-H04 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multip...lication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-119 ...

  12. Arabidopsis CDS blastp result: AK061395 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061395 006-305-E02 At2g02180.1 tobamovirus multiplication protein 3 (TOM3) identical to tobamovirus multip...lication protein (TOM3) GI:15425641 from [Arabidopsis thaliana] 1e-125 ...

  13. Arabidopsis CDS blastp result: AK243481 [KOME

    Lifescience Database Archive (English)

    Full Text Available rylethanolamine transferase, putative / CTP:phosphoethanolamine cytidylyltransferas...AK243481 J100072P04 At2g38670.1 68415.m04749 ethanolamine-phosphate cytidylyltransferase, putative / phospho

  14. Arabidopsis CDS blastp result: AK099943 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099943 J013121N21 At2g38670.1 ethanolamine-phosphate cytidylyltransferase, putative / phosphorylethanolami...ne transferase, putative / CTP:phosphoethanolamine cytidylyltransferase, putative s

  15. Arabidopsis CDS blastp result: AK288229 [KOME

    Lifescience Database Archive (English)

    Full Text Available rylethanolamine transferase, putative / CTP:phosphoethanolamine cytidylyltransferas...AK288229 J090012M05 At2g38670.1 68415.m04749 ethanolamine-phosphate cytidylyltransferase, putative / phospho

  16. Arabidopsis CDS blastp result: AK099644 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK099644 J013062A17 At2g38670.1 ethanolamine-phosphate cytidylyltransferase, putative / phosphorylethanolami...ne transferase, putative / CTP:phosphoethanolamine cytidylyltransferase, putative s

  17. Arabidopsis CDS blastp result: AK068868 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK068868 J023002G17 At2g38670.1 ethanolamine-phosphate cytidylyltransferase, putative / phosphorylethanolami...ne transferase, putative / CTP:phosphoethanolamine cytidylyltransferase, putative s

  18. Arabidopsis CDS blastp result: AK240726 [KOME

    Lifescience Database Archive (English)

    Full Text Available rylethanolamine transferase, putative / CTP:phosphoethanolamine cytidylyltransferas...AK240726 J043027P03 At2g38670.1 68415.m04749 ethanolamine-phosphate cytidylyltransferase, putative / phospho

  19. Arabidopsis CDS blastp result: AK065277 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065277 J013002L14 At1g12520.1 superoxide dismutase copper chaperone, putative sim...ilar to copper chaperone for superoxide dismutase [Homo sapiens] gi|2431868|gb|AAC51764 4e-88 ...

  20. Arabidopsis CDS blastp result: AK071224 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK071224 J023087B12 At5g55810.1 nicotinamide-nucleotide adenylyltransferase, putati...ve / NAD(+) pyrophosphorylase, putative similar to nicotinamide mononucleotide adenylyl transferase [Homo sa

  1. Arabidopsis CDS blastp result: AK242265 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242265 J075185M21 At5g55810.1 68418.m06955 nicotinamide-nucleotide adenylyltransf...erase, putative / NAD(+) pyrophosphorylase, putative similar to nicotinamide mononucleotide adenylyl transfe

  2. Arabidopsis CDS blastp result: AK288092 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288092 J075185M21 At5g55810.1 68418.m06955 nicotinamide-nucleotide adenylyltransf...erase, putative / NAD(+) pyrophosphorylase, putative similar to nicotinamide mononucleotide adenylyl transfe

  3. UniProt search blastx result: AK287424 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287424 J043006K04 P41398|AK_CORFL Aspartokinase (EC 2.7.2.4) (Aspartate kinase) [Contains: Asparto...kinase subunit alpha; Aspartokinase subunit beta] - Corynebacterium flavum 1.00E-24 ...

  4. Rbfox1 downregulation and altered calpain 3 splicing by FRG1 in a mouse model of Facioscapulohumeral muscular dystrophy (FSHD.

    Directory of Open Access Journals (Sweden)

    Mariaelena Pistoni

    Full Text Available Facioscapulohumeral muscular dystrophy (FSHD is a common muscle disease whose molecular pathogenesis remains largely unknown. Over-expression of FSHD region gene 1 (FRG1 in mice, frogs, and worms perturbs muscle development and causes FSHD-like phenotypes. FRG1 has been implicated in splicing, and we asked how splicing might be involved in FSHD by conducting a genome-wide analysis in FRG1 mice. We find that splicing perturbations parallel the responses of different muscles to FRG1 over-expression and disease progression. Interestingly, binding sites for the Rbfox family of splicing factors are over-represented in a subset of FRG1-affected splicing events. Rbfox1 knockdown, over-expression, and RNA-IP confirm that these are direct Rbfox1 targets. We find that FRG1 is associated to the Rbfox1 RNA and decreases its stability. Consistent with this, Rbfox1 expression is down-regulated in mice and cells over-expressing FRG1 as well as in FSHD patients. Among the genes affected is Calpain 3, which is mutated in limb girdle muscular dystrophy, a disease phenotypically similar to FSHD. In FRG1 mice and FSHD patients, the Calpain 3 isoform lacking exon 6 (Capn3 E6- is increased. Finally, Rbfox1 knockdown and over-expression of Capn3 E6- inhibit muscle differentiation. Collectively, our results suggest that a component of FSHD pathogenesis may arise by over-expression of FRG1, reducing Rbfox1 levels and leading to aberrant expression of an altered Calpain 3 protein through dysregulated splicing.

  5. Rbfox1 downregulation and altered calpain 3 splicing by FRG1 in a mouse model of Facioscapulohumeral muscular dystrophy (FSHD).

    Science.gov (United States)

    Pistoni, Mariaelena; Shiue, Lily; Cline, Melissa S; Bortolanza, Sergia; Neguembor, Maria Victoria; Xynos, Alexandros; Ares, Manuel; Gabellini, Davide

    2013-01-01

    Facioscapulohumeral muscular dystrophy (FSHD) is a common muscle disease whose molecular pathogenesis remains largely unknown. Over-expression of FSHD region gene 1 (FRG1) in mice, frogs, and worms perturbs muscle development and causes FSHD-like phenotypes. FRG1 has been implicated in splicing, and we asked how splicing might be involved in FSHD by conducting a genome-wide analysis in FRG1 mice. We find that splicing perturbations parallel the responses of different muscles to FRG1 over-expression and disease progression. Interestingly, binding sites for the Rbfox family of splicing factors are over-represented in a subset of FRG1-affected splicing events. Rbfox1 knockdown, over-expression, and RNA-IP confirm that these are direct Rbfox1 targets. We find that FRG1 is associated to the Rbfox1 RNA and decreases its stability. Consistent with this, Rbfox1 expression is down-regulated in mice and cells over-expressing FRG1 as well as in FSHD patients. Among the genes affected is Calpain 3, which is mutated in limb girdle muscular dystrophy, a disease phenotypically similar to FSHD. In FRG1 mice and FSHD patients, the Calpain 3 isoform lacking exon 6 (Capn3 E6-) is increased. Finally, Rbfox1 knockdown and over-expression of Capn3 E6- inhibit muscle differentiation. Collectively, our results suggest that a component of FSHD pathogenesis may arise by over-expression of FRG1, reducing Rbfox1 levels and leading to aberrant expression of an altered Calpain 3 protein through dysregulated splicing.

  6. SwissProt search result: AK073189 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK073189 J033004I10 (P37142) Bifunctional aspartokinase/homoserine dehydrogenase, c...hloroplast precursor (AK-HD) (AK-HSDH) [Includes: Aspartokinase (EC 2.7.2.4); Homoserine dehydrogenase (EC 1.1.1.3)] (Fragment) AKH_DAUCA 3e-40 ...

  7. Molecular Shape Analysis-Guided Virtual Screening Platform for Adenosine Kinase Inhibitors.

    Science.gov (United States)

    Bhutoria, Savita; Das, Ballari; Ghoshal, Nanda

    2016-01-01

    We propose a new application of molecular shape descriptors in hierarchical selection during virtual screening (VS). Here, a structure-based pharmacophore and docking-guided VS protocol have been evolved to identify inhibitors against adenosine kinase (AK). The knowledge gained on the shape requirements has been extrapolated in classifying active and inactive molecules against this target. This classification enabled us to pick the appropriate ligand conformation in the binding site. We have suggested a set of hierarchical filters for VS, from a simple molecular shape analysis (MSA) descriptor-based recursive models to docking scores. This approach permits a systematic study to understand the importance of spatial requirements and limitations for inhibitors against AK. Finally, the guidelines on how to select compounds for AK to achieve success have been highlighted. The utility of this approach has been suggested by giving an example of database screening for plausible active compounds.

  8. Human U87 astrocytoma cell invasion induced by interaction of βig-h3 with integrin α5β1 involves calpain-2.

    Directory of Open Access Journals (Sweden)

    Jie Ma

    Full Text Available It is known that βig-h3 is involved in the invasive process of many types of tumors, but its mechanism in glioma cells has not been fully clarified. Using immunofluorescent double-staining and confocal imaging analysis, and co-immunoprecipitation assays, we found that βig-h3 co-localized with integrin α5β1 in U87 cells. We sought to elucidate the function of this interaction by performing cell invasion assays and gelatin zymography experiments. We found that siRNA knockdowns of βig-h3 and calpain-2 impaired cell invasion and MMP secretion. Moreover, βig-h3, integrins and calpain-2 are known to be regulated by Ca(2+, and they are also involved in tumor cell invasion. Therefore, we further investigated if calpain-2 was relevant to βig-h3-integrin α5β1 interaction to affect U87 cell invasion. Our data showed that βig-h3 co-localized with integrin α5β1 to enhance the invasion of U87 cells, and that calpain-2, is involved in this process, acting as a downstream molecule.

  9. Similar to spironolactone, oxymatrine is protective in aldosterone-induced cardiomyocyte injury via inhibition of calpain and apoptosis-inducing factor signaling.

    Directory of Open Access Journals (Sweden)

    Ting-Ting Xiao

    Full Text Available Accumulating evidence indicates that oxymatrine (OMT possesses variously pharmacological properties, especially on the cardiovascular system. We previously demonstrated that activated calpain/apoptosis-inducing factor (AIF-mediated pathway was the key molecular mechanism in aldosterone (ALD induces cardiomyocytes apoptosis. In the present study, we extended the experimentation by investigating the effect of OMT on cardiomyocytes exposed to ALD, as compared to spironolactone (Spiro, a classical ALD receptor antagonist. Cardiomyocytes were pre-incubated with OMT, Spiro or vehicle for 1 h, and then, cardiomyocytes were exposed to ALD 24 h. The cell injury was evaluated by 3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT assay and lactate dehydrogenase (LDH leakage ratio. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL assay, annexin V/PI staining, and relative caspase-3 activity assay. Furthermore, expression of pro-apoptotic proteins including truncated Bid (tBid, calpain and AIF were evaluated by western blot analysis. ALD stimulation increased cardiomyocytes apoptosis, caspase-3 activity and protein expression of calpain, tBid and AIF in the cytosol (p<0.05. Pre-incubated with cardiomyocytes injury and increased caspase-3 activity were significantly attenuated (p<0.05. Furthermore, OMT suppressed ALD-induced high expression of calpain and AIF. And these effects of OMT could be comparable to Spiro. These findings indicated that OMT might be a potential cardioprotective-agent against excessive ALD-induced cardiotoxicity, at least in part, mediated through inhibition of calpain/AIF signaling.

  10. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At4g23990.1 68417.m03448 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 1e-124 ...

  11. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g24000.1 68417.m03449 cellulose synthase family protein similar to cellulose... synthase from Gossypium hirsutum [gi:1706956], cellulose synthase-5 from Zea mays [gi:9622882] 5e-27 ...

  12. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g38190.1 68417.m05391 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-5 (gi:9622882) from Zea mays 0.0 ...

  13. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g24000.1 68417.m03449 cellulose synthase family protein similar to cellulose... synthase from Gossypium hirsutum [gi:1706956], cellulose synthase-5 from Zea mays [gi:9622882] 2e-27 ...

  14. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At1g32180.1 68414.m03958 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-9 (gi:9622890) from Zea mays 1e-126 ...

  15. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At4g38190.1 68417.m05391 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-5 (gi:9622882) from Zea mays 4e-27 ...

  16. Arabidopsis CDS blastp result: AK061162 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061162 006-209-A01 At2g32540.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 3e-35 ...

  17. Arabidopsis CDS blastp result: AK110534 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK110534 002-168-A07 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 1e-114 ...

  18. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At4g38190.1 68417.m05391 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-5 (gi:9622882) from Zea mays 8e-63 ...

  19. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At1g55850.1 68414.m06405 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 0.0 ...

  20. Arabidopsis CDS blastp result: AK111344 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK111344 002-181-F12 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 2e-15 ...

  1. Arabidopsis CDS blastp result: AK243353 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243353 J100061B04 At1g05990.1 68414.m00627 calcium-binding protein, putative strong similarity to calcium...-binding protein [Lotus japonicus] GI:18413495; contains INTERPRO:IPR002048 calcium-binding EF-hand domain 3e-11 ...

  2. Arabidopsis CDS blastp result: AK066468 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066468 J013066B01 At1g53210.1 sodium/calcium exchanger family protein / calcium-b...inding EF hand family protein contains Pfam profiles: PF01699 sodium/calcium exchanger protein, PF00036 EF hand 1e-171 ...

  3. Arabidopsis CDS blastp result: AK071633 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK071633 J023102J23 At1g07080.1 gamma interferon responsive lysosomal thiol reducta...se family protein / GILT family protein similar to SP|P13284 Gamma-interferon inducible lysosomal thiol redu...ctase precursor {Homo sapiens}; contains Pfam profile PF03227: Gamma interferon inducible lysosomal thiol reductase (GILT) 6e-66 ...

  4. Arabidopsis CDS blastp result: AK106050 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK106050 001-206-F01 At1g07080.1 gamma interferon responsive lysosomal thiol reduct...ase family protein / GILT family protein similar to SP|P13284 Gamma-interferon inducible lysosomal thiol red...uctase precursor {Homo sapiens}; contains Pfam profile PF03227: Gamma interferon inducible lysosomal thiol reductase (GILT) 6e-63 ...

  5. Arabidopsis CDS blastp result: AK108564 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK108564 002-145-C08 At4g28088.1 hydrophobic protein, putative / low temperature an...d salt responsive protein, putative similar to Hydrophobic protein RCI2A (Low temperature and salt responsive protein LTI6A) GI:15214251 4e-11 ...

  6. Arabidopsis CDS blastp result: AK062410 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK062410 001-102-E12 At4g28088.1 hydrophobic protein, putative / low temperature an...d salt responsive protein, putative similar to Hydrophobic protein RCI2A (Low temperature and salt responsive protein LTI6A) GI:15214251 1e-13 ...

  7. Arabidopsis CDS blastp result: AK121018 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121018 J023049B21 At4g28088.1 hydrophobic protein, putative / low temperature and... salt responsive protein, putative similar to Hydrophobic protein RCI2A (Low temperature and salt responsive protein LTI6A) GI:15214251 2e-17 ...

  8. Arabidopsis CDS blastp result: AK242428 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242428 J080089P09 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 2e-14 ...

  9. Arabidopsis CDS blastp result: AK288095 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288095 J075191E21 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 2e-16 ...

  10. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At2g41100.2 68415.m05077 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 3e-44 ...

  11. Arabidopsis CDS blastp result: AK242428 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242428 J080089P09 At2g41100.2 68415.m05077 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 3e-16 ...

  12. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At5g37770.1 68418.m04547 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 2e-25 ...

  13. Arabidopsis CDS blastp result: AK062711 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK062711 001-106-C02 At5g37770.1 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 9e-34 ...

  14. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 8e-44 ...

  15. Arabidopsis CDS blastp result: AK108506 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK108506 002-143-H11 At5g37770.1 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 7e-14 ...

  16. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 3e-26 ...

  17. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At2g41100.2 68415.m05077 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 3e-26 ...

  18. Arabidopsis CDS blastp result: AK242428 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242428 J080089P09 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 8e-18 ...

  19. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At5g37770.1 68418.m04547 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 2e-11 ...

  20. Arabidopsis CDS blastp result: AK243656 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243656 J100088L22 At2g41100.2 68415.m05077 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 5e-20 ...

  1. Arabidopsis CDS blastp result: AK242346 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242346 J080012M07 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 4e-41 ...

  2. Arabidopsis CDS blastp result: AK243656 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243656 J100088L22 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 2e-17 ...

  3. Arabidopsis CDS blastp result: AK071661 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK071661 J023105D07 At5g37770.1 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 3e-33 ...

  4. Arabidopsis CDS blastp result: AK242428 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242428 J080089P09 At5g37770.1 68418.m04547 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 9e-19 ...

  5. Arabidopsis CDS blastp result: AK288095 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288095 J075191E21 At2g41100.2 68415.m05077 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 2e-15 ...

  6. Arabidopsis CDS blastp result: AK241786 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241786 J065207F05 At5g37770.1 68418.m04547 touch-responsive protein / calmodulin-related protein 2, touch...-induced (TCH2) identical to calmodulin-related protein 2,touch-induced SP:P25070 from [Arabidopsis thaliana] 1e-19 ...

  7. Arabidopsis CDS blastp result: AK243656 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243656 J100088L22 At2g41100.1 68415.m05076 touch-responsive protein / calmodulin-related protein 3, touch...-induced (TCH3) identical to calmodulin-related protein 3, touch-induced SP:P25071 from [Arabidopsis thaliana] 1e-19 ...

  8. Arabidopsis CDS blastp result: AK060920 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060920 001-035-H02 At3g55440.1 triosephosphate isomerase, cytosolic, putative strong similarity to triosep...hosphate isomerase, cytosolic from Petunia hybrida [SP|P48495], from Coptis japonica [SP|P21820] 1e-113 ...

  9. Arabidopsis CDS blastp result: AK104017 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104017 001-007-G07 At3g55440.1 triosephosphate isomerase, cytosolic, putative strong similarity to triosep...hosphate isomerase, cytosolic from Petunia hybrida [SP|P48495], from Coptis japonica [SP|P21820] 1e-106 ...

  10. Arabidopsis CDS blastp result: AK100708 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100708 J023115C24 At3g55440.1 triosephosphate isomerase, cytosolic, putative strong similarity to trioseph...osphate isomerase, cytosolic from Petunia hybrida [SP|P48495], from Coptis japonica [SP|P21820] 1e-106 ...

  11. Arabidopsis CDS blastp result: AK104461 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104461 006-301-B11 At3g55440.1 triosephosphate isomerase, cytosolic, putative strong similarity to triosep...hosphate isomerase, cytosolic from Petunia hybrida [SP|P48495], from Coptis japonica [SP|P21820] 1e-106 ...

  12. Arabidopsis CDS blastp result: AK107494 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK107494 002-129-D06 At4g24620.1 glucose-6-phosphate isomerase, putative similar to glucose...-6-phosphate isomerase [Spinacia oleracea] GI:3413511; contains Pfam profile PF00342: glucose-6-phosphate isomerase 0.0 ...

  13. Arabidopsis CDS blastp result: AK241096 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241096 J065076O13 At2g16060.1 68415.m01841 non-symbiotic hemoglobin 1 (HB1) (GLB1...) identical to SP|O24520 Non-symbiotic hemoglobin 1 (Hb1) (ARAth GLB1) {Arabidopsis thaliana} 1e-59 ...

  14. Arabidopsis CDS blastp result: AK240885 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240885 J065029A17 At3g10520.1 68416.m01262 non-symbiotic hemoglobin 2 (HB2) (GLB2...) identical to SP|O24521 Non-symbiotic hemoglobin 2 (Hb2) (ARAth GLB2) {Arabidopsis thaliana} 6e-34 ...

  15. Arabidopsis CDS blastp result: AK241096 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241096 J065076O13 At3g10520.1 68416.m01262 non-symbiotic hemoglobin 2 (HB2) (GLB2...) identical to SP|O24521 Non-symbiotic hemoglobin 2 (Hb2) (ARAth GLB2) {Arabidopsis thaliana} 1e-40 ...

  16. Arabidopsis CDS blastp result: AK240885 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240885 J065029A17 At2g16060.1 68415.m01841 non-symbiotic hemoglobin 1 (HB1) (GLB1...) identical to SP|O24520 Non-symbiotic hemoglobin 1 (Hb1) (ARAth GLB1) {Arabidopsis thaliana} 3e-49 ...

  17. Arabidopsis CDS blastp result: AK288081 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288081 J075172F18 At5g24520.3 68418.m02893 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 4e-13 ...

  18. Arabidopsis CDS blastp result: AK243285 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243285 J100051N01 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 1e-24 ...

  19. Arabidopsis CDS blastp result: AK243061 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243061 J100014C18 At5g24520.3 68418.m02893 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 1e-102 ...

  20. Arabidopsis CDS blastp result: AK243061 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243061 J100014C18 At5g24520.2 68418.m02892 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 1e-102 ...

  1. Arabidopsis CDS blastp result: AK289209 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK289209 J100058I16 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 1e-12 ...

  2. Arabidopsis CDS blastp result: AK287566 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287566 J065027L04 At1g34790.1 68414.m04337 transparent testa 1 protein (TT1) / zi...nc finger (C2H2 type) protein TT1 identical to transparent testa 1 GI:18253279 from [Arabidopsis thaliana]; contains Pfam profile PF00096: Zinc finger, C2H2 type 2e-77 ...

  3. Arabidopsis CDS blastp result: AK243061 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243061 J100014C18 At5g24520.1 68418.m02891 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 1e-102 ...

  4. Arabidopsis CDS blastp result: AK288081 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288081 J075172F18 At5g24520.2 68418.m02892 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 4e-13 ...

  5. Arabidopsis CDS blastp result: AK288081 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288081 J075172F18 At5g24520.1 68418.m02891 transparent testa glabra 1 protein (TTG1) identical to transpar...ent testa glabra 1 (Ttg1) protein (GI:10177852) {Arabidopsis thaliana}; contains Pfam PF00400: WD domain, G-beta repeat (4 copies,1 weak); 4e-13 ...

  6. Arabidopsis CDS blastp result: AK101554 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101554 J033050A06 At4g09020.1 isoamylase, putative / starch debranching enzyme, putative similar to isoamy...lase isoform 3 [Solanum tuberosum] GI:27728149, isoamylase [Oryza sativa] GI:325279...4; contains Pfam profiles PF00128: Alpha amylase catalytic domain, PF02922: Isoamylase N-terminal domain 1e-135 ...

  7. Arabidopsis CDS blastp result: AK242137 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242137 J075150A13 At4g09020.1 68417.m01489 isoamylase, putative / starch debranch...ing enzyme, putative similar to isoamylase isoform 3 [Solanum tuberosum] GI:27728149, isoamylase [Oryza sati...va] GI:3252794; contains Pfam profiles PF00128: Alpha amylase catalytic domain, PF02922: Isoamylase N-terminal domain 9e-13 ...

  8. Arabidopsis CDS blastp result: AK288307 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288307 J090020G01 At4g09020.1 68417.m01489 isoamylase, putative / starch debranch...ing enzyme, putative similar to isoamylase isoform 3 [Solanum tuberosum] GI:27728149, isoamylase [Oryza sati...va] GI:3252794; contains Pfam profiles PF00128: Alpha amylase catalytic domain, PF02922: Isoamylase N-terminal domain 8e-98 ...

  9. Arabidopsis CDS blastp result: AK100867 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100867 J023124E13 At2g29640.1 josephin family protein contains Pfam domain PF02099: Jose...phin; similar to Josephin-like protein (Swiss-Prot:O82391) [Arabidopsis thaliana] 7e-59 ...

  10. Arabidopsis CDS blastp result: AK069900 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK069900 J023035E20 At5g23420.1 high mobility group (HMG1/2) family protein similar to high... mobility group protein 2 HMG2 [Ipomoea nil] GI:1052956; contains Pfam profile PF00505: HMG (high mobility group) box 8e-32 ...

  11. Arabidopsis CDS blastp result: AK059151 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059151 001-023-B10 At4g25080.3 magnesium-protoporphyrin O-methyltransferase, putative / magnesium-protopor...phyrin IX methyltransferase, putative similar to SP|Q55467 Magnesium-protoporphyrin... O-methyltransferase (EC 2.1.1.1) (Magnesium-protoporphyrin IX methyltransferase) {Synechocystis sp.} 4e-31 ...

  12. Arabidopsis CDS blastp result: AK066771 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK066771 J013083K07 At1g01170.1 ozone-responsive stress-related protein, putative s...imilar to stress-related ozone-induced protein AtOZI1 (GI:790583) [Arabidopsis thaliana]; contains 1 predicted transmembrane domain; 2e-29 ...

  13. Arabidopsis CDS blastp result: AK059353 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059353 001-026-D01 At1g01170.1 ozone-responsive stress-related protein, putative ...similar to stress-related ozone-induced protein AtOZI1 (GI:790583) [Arabidopsis thaliana]; contains 1 predicted transmembrane domain; 2e-29 ...

  14. Arabidopsis CDS blastp result: AK059160 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK059160 001-023-D05 At1g01170.1 ozone-responsive stress-related protein, putative ...similar to stress-related ozone-induced protein AtOZI1 (GI:790583) [Arabidopsis thaliana]; contains 1 predicted transmembrane domain; 3e-28 ...

  15. Arabidopsis CDS blastp result: AK110681 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK110681 002-169-H04 At5g09650.1 inorganic pyrophosphatase family protein similar to SP|Q15181 Inorganic... pyrophosphatase (EC 3.6.1.1) (Pyrophosphate {Homo sapiens}; contains Pfam profile PF00719: inorganic pyrophosphatase 8e-63 ...

  16. Arabidopsis CDS blastp result: AK069313 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK069313 J023014E07 At3g53620.1 inorganic pyrophosphatase, putative [soluble] / pyr...ophosphate phospho-hydrolase, putative / PPase, putative similar to magnesium dependent soluble inorganic py...rophosphatase [Solanum tuberosum] GI:2706450; contains Pfam profile PF00719: inorganic pyrophosphatase 1e-105 ...

  17. Arabidopsis CDS blastp result: AK242307 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242307 J075194A02 At1g78920.1 68414.m09201 vacuolar-type H+-translocating inorganic... pyrophosphatase (AVPL1) identical to vacuolar-type H+-translocating inorganic pyrophosphatase GI:6901676 from [Arabidopsis thaliana] 5e-41 ...

  18. Arabidopsis CDS blastp result: AK070310 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070310 J023047F16 At1g16780.1 vacuolar-type H+-translocating inorganic pyrophosph...atase, putative similar to vacuolar-type H+-translocating inorganic pyrophosphatase GI:6901676 from [Arabidopsis thaliana] 0.0 ...

  19. Arabidopsis CDS blastp result: AK242307 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242307 J075194A02 At1g16780.1 68414.m02016 vacuolar-type H+-translocating inorganic... pyrophosphatase, putative similar to vacuolar-type H+-translocating inorganic pyrophosphatase GI:6901676 from [Arabidopsis thaliana] 2e-40 ...

  20. Arabidopsis CDS blastp result: AK241684 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241684 J065194D01 At3g53620.1 68416.m05923 inorganic pyrophosphatase, putative [s...oluble] / pyrophosphate phospho-hydrolase, putative / PPase, putative similar to magnesium dependent soluble inorganic... pyrophosphatase [Solanum tuberosum] GI:2706450; contains Pfam profile PF00719: inorganic pyrophosphatase 1e-105 ...

  1. Arabidopsis CDS blastp result: AK241112 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241112 J065091K02 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 1e-14 ...

  2. Arabidopsis CDS blastp result: AK288612 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288612 J090053J15 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 5e-24 ...

  3. Arabidopsis CDS blastp result: AK287737 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287737 J065143M09 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 7e-14 ...

  4. Arabidopsis CDS blastp result: AK287434 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287434 J043012F24 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 2e-27 ...

  5. Arabidopsis CDS blastp result: AK241112 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241112 J065091K02 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 3e-13 ...

  6. Arabidopsis CDS blastp result: AK240855 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240855 J065021H02 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 7e-25 ...

  7. Arabidopsis CDS blastp result: AK289251 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK289251 J100081E23 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 6e-21 ...

  8. Arabidopsis CDS blastp result: AK241112 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241112 J065091K02 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 1e-16 ...

  9. Arabidopsis CDS blastp result: AK241784 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241784 J065206N09 At4g16390.1 68417.m02481 chloroplastic RNA-binding protein P67,... putative nearly identical to 67kD chloroplastic RNA-binding protein, P67 [Arabidopsis thaliana] GI:9755842 4e-11 ...

  10. Arabidopsis CDS blastp result: AK121784 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121784 J033094A11 At2g21270.1 ubiquitin fusion degradation UFD1 family protein si...milar to SP|P70362 Ubiquitin fusion degradation protein 1 homolog (UB fusion protein 1) {Mus musculus}; cont...ains Pfam profile PF03152: Ubiquitin fusion degradation protein UFD1 1e-106 ...

  11. Arabidopsis CDS blastp result: AK120576 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120576 J013135C24 At2g21270.1 ubiquitin fusion degradation UFD1 family protein si...milar to SP|P70362 Ubiquitin fusion degradation protein 1 homolog (UB fusion protein 1) {Mus musculus}; contains Pfam profile PF03152: Ubiquitin fusion degradation protein UFD1 4e-89 ...

  12. Arabidopsis CDS blastp result: AK119957 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK119957 002-184-C12 At2g21270.1 ubiquitin fusion degradation UFD1 family protein s...imilar to SP|P70362 Ubiquitin fusion degradation protein 1 homolog (UB fusion protein 1) {Mus musculus}; con...tains Pfam profile PF03152: Ubiquitin fusion degradation protein UFD1 1e-96 ...

  13. Arabidopsis CDS blastp result: AK073711 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK073711 J033061G23 At2g21270.1 ubiquitin fusion degradation UFD1 family protein si...milar to SP|P70362 Ubiquitin fusion degradation protein 1 homolog (UB fusion protein 1) {Mus musculus}; cont...ains Pfam profile PF03152: Ubiquitin fusion degradation protein UFD1 1e-110 ...

  14. Arabidopsis CDS blastp result: AK102921 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102921 J033114C10 At4g15770.1 60S ribosome subunit biogenesis protein, putative c...ontains similarity to 60S ribosome subunit biogenesis protein NIP7 (Swiss-Prot:Q08962) [Saccharomyces cerevisiae] 4e-26 ...

  15. Arabidopsis CDS blastp result: AK060349 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060349 001-008-F05 At4g15770.1 60S ribosome subunit biogenesis protein, putative ...contains similarity to 60S ribosome subunit biogenesis protein NIP7 (Swiss-Prot:Q08962) [Saccharomyces cerevisiae] 1e-29 ...

  16. Arabidopsis CDS blastp result: AK243057 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243057 J100013P13 At3g62060.1 68416.m06973 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GI:1431629 from [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-103 ...

  17. Arabidopsis CDS blastp result: AK243057 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243057 J100013P13 At3g09410.3 68416.m01119 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GB:CAA67728 [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-106 ...

  18. Arabidopsis CDS blastp result: AK243057 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243057 J100013P13 At3g09410.2 68416.m01117 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GB:CAA67728 [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 2e-96 ...

  19. Arabidopsis CDS blastp result: AK243057 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK243057 J100013P13 At3g09410.1 68416.m01118 pectinacetylesterase family protein similar to pectin...acetylesterase precursor GB:CAA67728 [Vigna radiata]; contains Pfam profile: PF03283 pectinacetylesterase 1e-106 ...

  20. Arabidopsis CDS blastp result: AK240821 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240821 J065011J02 At3g16360.1 68416.m02070 phosphotransfer family protein similar to two-component phospho...relay mediators ATHP1 (GI:4156241), ATHP3 (GI:4156245) [Arabidopsis thaliana], hist

  1. Arabidopsis CDS blastp result: AK242899 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242899 J090081K17 At5g01670.1 68418.m00083 aldose reductase, putative similar to aldose... reductase [Hordeum vulgare][GI:728592], aldose reductase ALDRXV4 [Xerophyta viscosa][GI:4539944] 9e-66 ...

  2. Arabidopsis CDS blastp result: AK242899 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242899 J090081K17 At5g01670.2 68418.m00084 aldose reductase, putative similar to aldose... reductase [Hordeum vulgare][GI:728592], aldose reductase ALDRXV4 [Xerophyta viscosa][GI:4539944] 2e-61 ...

  3. Arabidopsis CDS blastp result: AK102864 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102864 J033111K17 At5g01670.1 aldose reductase, putative similar to aldose reduct...ase [Hordeum vulgare][GI:728592], aldose reductase ALDRXV4 [Xerophyta viscosa][GI:4539944] 1e-120 ...

  4. Arabidopsis CDS blastp result: AK107208 [KOME

    Lifescience Database Archive (English)

    Full Text Available Ala hydrolase, putative virtually identical to gr1-protein from [Arabidopsis thaliana] GI:3559811; similar t...AK107208 002-125-B11 At1g44350.1 IAA-amino acid hydrolase 6, putative (ILL6) / IAA-

  5. Arabidopsis CDS blastp result: AK287447 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287447 J043016O04 At3g61850.2 68416.m06946 Dof zinc finger protein DAG1 / Dof affect...ing germination 1 (DAG1) / transcription factor BBFa (BBFA) identical to SP|Q43385 DOF zinc finger protein DAG1 (Dof affect

  6. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At3g06500.1 68416.m00754 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  7. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At5g22510.1 68418.m02627 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  8. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At1g35580.2 68414.m04418 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-164 ...

  9. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At4g34860.1 68417.m04945 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-164 ...

  10. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At1g22650.1 68414.m02830 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-159 ...

  11. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At1g35580.1 68414.m04417 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-164 ...

  12. Arabidopsis CDS blastp result: AK103334 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103334 J033125P15 At1g56560.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  13. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At3g05820.1 68416.m00653 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  14. Arabidopsis CDS blastp result: AK100373 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK100373 J023084J15 At5g22510.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-128 ...

  15. Arabidopsis CDS blastp result: AK120720 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120720 J023002G12 At1g56560.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  16. Arabidopsis CDS blastp result: AK102741 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102741 J033106F23 At4g09510.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  17. Arabidopsis CDS blastp result: AK061240 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061240 006-211-C06 At1g56560.1 beta-fructofuranosidase, putative / invertase, put...ative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-133 ...

  18. Arabidopsis CDS blastp result: AK070884 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK070884 J023068M15 At4g09510.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  19. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At4g09510.1 68417.m01563 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-164 ...

  20. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At1g56560.1 68414.m06505 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  1. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At1g72000.1 68414.m08322 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-155 ...

  2. Arabidopsis CDS blastp result: AK065562 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK065562 J013030A21 At4g09510.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  3. Arabidopsis CDS blastp result: AK288558 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK288558 J090047H18 At4g09510.2 68417.m01564 beta-fructofuranosidase, putative / invertase..., putative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus ...carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 1e-135 ...

  4. Arabidopsis CDS blastp result: AK121301 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121301 J023110M10 At5g22510.1 beta-fructofuranosidase, putative / invertase, puta...tive / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 0.0 ...

  5. Arabidopsis CDS blastp result: AK061496 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061496 006-309-D01 At1g22650.1 beta-fructofuranosidase, putative / invertase, put...ative / saccharase, putative / beta-fructosidase, putative similar to neutral invertase [Daucus carota] GI:4200165; contains Pfam profile PF04853: Plant neutral invertase 2e-88 ...

  6. Arabidopsis CDS blastp result: AK240924 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240924 J065039N21 At3g16000.1 68416.m02024 matrix-localized MAR DNA-binding prote...in-related similar to matrix-localized MAR DNA binding protein MFP1 GI:1771158 from [Lycopersicon esculentum] 1e-96 ...

  7. Arabidopsis CDS blastp result: AK241886 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK241886 J065219F05 At1g75280.1 68414.m08745 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 5e-67 ...

  8. Arabidopsis CDS blastp result: AK287747 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK287747 J065152G10 At1g75280.1 68414.m08745 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 4e-36 ...

  9. Arabidopsis CDS blastp result: AK098946 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK098946 J013045H04 At1g75280.1 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 5e-85 ...

  10. Arabidopsis CDS blastp result: AK104669 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104669 006-312-B02 At1g75280.1 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 5e-85 ...

  11. Arabidopsis CDS blastp result: AK061325 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061325 006-302-G05 At1g75280.1 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 5e-85 ...

  12. Arabidopsis CDS blastp result: AK102701 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102701 J033103L17 At1g75280.1 isoflavone reductase, putative identical to SP|P52577 Isoflavon...e reductase homolog P3 (EC 1.3.1.-) {Arabidopsis thaliana}; contains Pfam profile PF02716: isoflavone reductase 9e-93 ...

  13. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At1g32180.1 68414.m03958 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-9 (gi:9622890) from Zea mays 1e-24 ...

  14. Arabidopsis CDS blastp result: AK109812 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK109812 002-147-H02 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 5e-90 ...

  15. Arabidopsis CDS blastp result: AK101487 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK101487 J033042D19 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 0.0 ...

  16. Arabidopsis CDS blastp result: AK105393 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK105393 001-123-B04 At5g16910.1 cellulose synthase family protein similar to gi:2827143 cellulose... synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  17. Arabidopsis CDS blastp result: AK120054 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK120054 J013000L05 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 1e-148 ...

  18. Arabidopsis CDS blastp result: AK121003 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK121003 J023045B21 At2g32540.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 1e-167 ...

  19. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At4g23990.1 68417.m03448 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 1e-45 ...

  20. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g32410.1 68417.m04614 cellulose synthase, catalytic subunit, putative similar to cell...ulose synthase-1 [gi:9622874] and -2 [gi:9622876] from Zea mays 0.0 ...

  1. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g23990.1 68417.m03448 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 8e-25 ...

  2. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At2g32540.1 68415.m03975 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 4e-98 ...

  3. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At4g24000.1 68417.m03449 cellulose synthase family protein similar to cellulose... synthase from Gossypium hirsutum [gi:1706956], cellulose synthase-5 from Zea mays [gi:9622882] 4e-48 ...

  4. Arabidopsis CDS blastp result: AK061639 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK061639 001-036-B01 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 4e-49 ...

  5. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At4g32410.1 68417.m04614 cellulose synthase, catalytic subunit, putative similar to cell...ulose synthase-1 [gi:9622874] and -2 [gi:9622876] from Zea mays 1e-105 ...

  6. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At2g32540.1 68415.m03975 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 2e-45 ...

  7. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At2g32540.1 68415.m03975 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 3e-31 ...

  8. Arabidopsis CDS blastp result: AK060286 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK060286 001-006-C08 At2g32540.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 6e-78 ...

  9. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At1g02730.1 68414.m00226 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-4 [gi:9622880] from Zea mays 7e-27 ...

  10. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At1g32180.1 68414.m03958 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-9 (gi:9622890) from Zea mays 3e-66 ...

  11. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At4g24000.1 68417.m03449 cellulose synthase family protein similar to cellulose... synthase from Gossypium hirsutum [gi:1706956], cellulose synthase-5 from Zea mays [gi:9622882] 1e-123 ...

  12. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At2g32530.1 68415.m03974 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 5e-48 ...

  13. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At2g32530.1 68415.m03974 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 2e-29 ...

  14. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g24000.1 68417.m03449 cellulose synthase family protein similar to cellulose... synthase from Gossypium hirsutum [gi:1706956], cellulose synthase-5 from Zea mays [gi:9622882] 4e-25 ...

  15. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At5g16910.1 68418.m01982 cellulose synthase family protein similar to gi:2827143 cellulo...se synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 1e-130 ...

  16. Arabidopsis CDS blastp result: AK067424 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK067424 J013107C16 At1g02730.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-4 [gi:9622880] from Zea mays 0.0 ...

  17. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At4g32410.1 68417.m04614 cellulose synthase, catalytic subunit, putative similar to cell...ulose synthase-1 [gi:9622874] and -2 [gi:9622876] from Zea mays 6e-43 ...

  18. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At1g55850.1 68414.m06405 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 2e-22 ...

  19. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At2g32530.1 68415.m03974 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 8e-98 ...

  20. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g23990.1 68417.m03448 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 2e-26 ...

  1. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At1g02730.1 68414.m00226 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-4 [gi:9622880] from Zea mays 1e-69 ...

  2. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At5g44030.1 68418.m05388 cellulose synthase, catalytic subunit ...(IRX5) nearly identical to cellulose synthase [Arabidopsis thaliana] GI:27462651; contains Pfam profile PF03552: Cellulose synthase 6e-54 ...

  3. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At1g55850.1 68414.m06405 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 1e-52 ...

  4. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At1g02730.1 68414.m00226 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-4 [gi:9622880] from Zea mays 1e-131 ...

  5. Arabidopsis CDS blastp result: AK102766 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK102766 J033107E04 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 0.0 ...

  6. Arabidopsis CDS blastp result: AK107881 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK107881 002-134-D06 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 5e-51 ...

  7. Arabidopsis CDS blastp result: AK069071 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK069071 J023010H01 At2g32540.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 1e-167 ...

  8. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At2g32540.1 68415.m03975 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 4e-47 ...

  9. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At1g02730.1 68414.m00226 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-4 [gi:9622880] from Zea mays 0.0 ...

  10. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At5g16910.1 68418.m01982 cellulose synthase family protein similar to gi:2827143 cellulo...se synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 1e-28 ...

  11. Arabidopsis CDS blastp result: AK103810 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK103810 J033147A19 At1g55850.1 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 1e-179 ...

  12. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At2g32530.1 68415.m03974 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 4e-50 ...

  13. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At5g16910.1 68418.m01982 cellulose synthase family protein similar to gi:2827143 cellulo...se synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 0.0 ...

  14. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At5g16910.1 68418.m01982 cellulose synthase family protein similar to gi:2827143 cellulo...se synthase catalytic subunit, Arabidopsis thaliana, gi:9622886 cellulose synthase-7 from Zea mays 2e-65 ...

  15. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At4g23990.1 68417.m03448 cellulose synthase family protein similar to cellulose... synthase catalytic subunit from Arabidopsis thaliana [gi:5230423], cellulose synthase-5 from Zea mays [gi:9622882] 5e-25 ...

  16. Arabidopsis CDS blastp result: AK242890 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242890 J090079L19 At4g38190.1 68417.m05391 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-5 (gi:9622882) from Zea mays 1e-125 ...

  17. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At1g55850.1 68414.m06405 cellulose synthase family protein similar to cellulose... synthase catalytic subunit [gi:13925881] from Nicotiana alata, cellulose synthase-5 [gi:9622882] from Zea mays 1e-61 ...

  18. Arabidopsis CDS blastp result: AK242601 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242601 J090014G03 At1g32180.1 68414.m03958 cellulose synthase family protein similar to cellulose... synthase catalytic subunit gi:2827143 from [Arabidopsis thaliana], cellulose synthase-9 (gi:9622890) from Zea mays 0.0 ...

  19. Arabidopsis CDS blastp result: AK242585 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK242585 J090010M20 At5g44030.1 68418.m05388 cellulose synthase, catalytic subunit ...(IRX5) nearly identical to cellulose synthase [Arabidopsis thaliana] GI:27462651; contains Pfam profile PF03552: Cellulose synthase 2e-29 ...

  20. Arabidopsis CDS blastp result: AK072735 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK072735 J023141D23 At3g05000.1 transport protein particle (TRAPP) component Bet3 f...amily protein similar to Transport protein particle 33 kDa subunit (TRAPP 33 kDa subunit) (Swiss-Prot:Q99394...) [Saccharomyces cerevisiae]; contains Pfam profile PF04051: Transport protein particle (TRAPP) component, Bet3 1e-49 ...