Mutagenesis of the bovSERPINA3‐3 demonstrates the requirement of aspartate‐371 for intermolecular interaction and formation of dimers

Blanchet, Xavier; Péré-Brissaud, A; Duprat, Nathalie; Pinault, Émilie; Delourme, Didier; Ouali, Ahmed; Combet, Christophe; Maftah, Abderrahman; Pélissier, Patrick; Brémaud, Laure

doi:10.1002/pro.2078

Cited by 7 publications

(13 citation statements)

References 46 publications

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“…Trypsin is similarly inhibited by the recombinant WT and the M-D 37 /A 37 bovSERPINA3-3, and both mutated and WT serpins formed a SDS-stable complex with the serine protease. By contrast, caspase 3 is no more inhibited with the mutant M-D 37 /A 37 bovSERPINA3-3 as compared to the wild-type recombinant protein and, as expected, SDS-stable complex was obtained for the WT but not for the mutated serpin [8,35]. This allows us to conclude that the P1 residue for caspases is very likely Asp 37 or Asp 371 in the native protein.…”

Section: Nature and Localization Of The Target Scissile Bond Within Tsupporting

confidence: 72%

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Caspases and Thrombin Activity Regulation by Specific Serpin Inhibitors in Bovine Skeletal Muscle

Gagaoua

Hafid

Boudida

et al. 2015

Appl Biochem Biotechnol

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In living cells, after activation, protein inhibitors constitute the last step of proteases activity regulation. This review intends to provide original information about a group of bovine muscle serine proteases inhibitors belonging to the Serpin superfamily and characterized at the gene and protein level. This report is the only one and the first to provide much information on this group of proteases inhibitors of the serpin type and their potential biological functions. Amongst the eight genes identified in bovine, three serpins were purified from the muscle tissue and characterized. These are two members of the bovSERPINA3 family, i.e., bovSERPINA3-1 and A3-3, and the last one is antithrombin III (AT-III or BovSERPINC1). BovSERPINA3 family comprises at least eight protein members encoded by different genes mapped on chromosome 7q23-q26 cluster. BovSERPINA3-1 and A3-3 were shown to locate within muscle cells and are cross-class inhibitors strongly active against trypsin as well as against human initiator and effector caspases 8 and 3. They constitute a key apoptosis control in mammals. They were thus expressed in proliferating and confluent myoblasts phases where cells must be alive but not in myotubes. Antithrombin III inhibits trypsin and, in a heparin dependent manner, thrombin. AT-III and its mRNA were expressed in muscle cells and in differentiating primary myoblasts in culture.

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Section: Nature and Localization Of The Target Scissile Bond Within Tsupporting

confidence: 72%

“…In fact, as they were glycosylated [8] and phosphorylated to various extent, the number of isoforms is much larger. This assumption was confirmed by either one-or two-dimensional gel electrophoresis of fractionated muscle extracts [27,28].…”

Section: Cellular Localization Tissue Distribution and Polymorphismmentioning

confidence: 99%

Caspases and Thrombin Activity Regulation by Specific Serpin Inhibitors in Bovine Skeletal Muscle

Gagaoua

Hafid

Boudida

et al. 2015

Appl Biochem Biotechnol

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“…This was confirmed recently by PNGase F (progressive removal of all N-glycans) treatment of recombinant glycosylated bovSERPINA3-3 produced and purified from S. cerevisiae [26]. Such treatment revealed six states of glycosylation corresponding to six different forms of different Mr separated by SDS-PAGE.…”

Section: Polymorphism Of Bovserpina3s At the Protein Levelsupporting

confidence: 63%

New Caspases’ inhibitors belonging to the serpin superfamily: A novel key control point of apoptosis in mammalian tissues

Gagaoua¹,

Boudida²,

Becila³

et al. 2012

ABB

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The present report overviews a new family of bovine serpins able to inhibit pseudo-irreversibly initiator and effector caspases, a group of cysteine proteases in charge of cell dismantling during apoptosis, a finely regulated cell death process. The 8 members identified at the gene level showed a high homology with human SERPINA3 and were therefore designed bovSERPINA3-1 to A3-8. At least six of them are able to inhibit caspases. Two of them (bovSERPINA3-1 and A3-3) have been purified from bovine muscle and extensively investigated during these last years. After a general presentation of the serpin superfamily, the kinetic aspects of their interaction with human caspases 3 and 8 were studied and findings obtained suggest that caspases could be their target enzymes in living cells. In muscle and primary myoblast in culture, they showed an intracellular localization and because of their high level in blood, they can be exported. Two biological functions (potential regulator of apoptosis and expression during myoblast differentiation) were investigated and it was concluded that they are very likely an efficient regulator of apoptosis, a proposal supported by their high expression in proliferating myoblast (cell survival is essential during this differentiation phase) but not in myotubes.

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“…Despite the addition of urea, neither Dithiothreitol (DTT) nor betamercaptoethanol was able to modify the upper band, but peptides that allow the identification of PILP-B1 were found in both the upper (100 kDa) and lower (55 kDa) band (IC samples), which strongly suggests that the same protein is present in both bands as they are recognized by the mAb. Unfortunately, the mechanism of serpin dimerization/polymerization is not known [35,38] but it is believed that this process occurs due to a domain swap mechanism [39].…”

Section: Discussionmentioning

confidence: 99%

MSP22.8 is a protease inhibitor‐like protein involved in shell mineralization in the edible mussel Mytilus galloprovincialis

Calvo-Iglesias

Pérez-Estévez²,

González-Fernández

2017

FEBS Open Bio

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The mussel shell protein 22.8 (MSP22.8) is recognized by a monoclonal antibody (M22.8) directed against larvae of the mussel Mytilus galloprovincialis. After being secreted by cells of the mantle‐edge epithelium into the extrapallial (EP) space (the gap between the mantle and the shell), the protein is detected in the extrapallial fluid (EPF) and EP hemocytes and finally becomes part of the shell matrix framework in adult specimens of M. galloprovincialis. In the work described here, we show how MSP22.8 is detected in EPF samples from different species of mussels (M. galloprovincialis, Mytilus edulis, and Xenostrobus securis), and also as a shell matrix protein in M. galloprovincialis, Mytilus chilensis, and Perna canaliculus. A multistep purification strategy was employed to isolate the protein from the EPF, which was then analyzed by mass spectrometry in order to identify it. The results indicate that MSP22.8 is a serpin‐like protein that has great similarity with the protease inhibitor‐like protein‐B1, reported previously for Mytilus coruscus. We suggest that MSP22.8 is part of a system offering protection from proteolysis during biomineralization and is also part of the innate immune system in mussels.

show abstract

Mutagenesis of the bovSERPINA3‐3 demonstrates the requirement of aspartate‐371 for intermolecular interaction and formation of dimers

Cited by 7 publications

References 46 publications

Caspases and Thrombin Activity Regulation by Specific Serpin Inhibitors in Bovine Skeletal Muscle

Caspases and Thrombin Activity Regulation by Specific Serpin Inhibitors in Bovine Skeletal Muscle

New Caspases’ inhibitors belonging to the serpin superfamily: A novel key control point of apoptosis in mammalian tissues

MSP22.8 is a protease inhibitor‐like protein involved in shell mineralization in the edible mussel Mytilus galloprovincialis

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