Functional mapping of factor VIII C2 domain
| Autor: | Marc Delcourt, Jean-Luc Pellequer, Jean-Luc Plantier, Didier Saboulard, Claude Negrier, Shu-wen W. Chen |
|---|---|
| Přispěvatelé: | Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service de Biochimie et Toxicologie Nucléaire (SBTN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), BIOMETHODES, Centre Hospitalier Universitaire de Lyon (CHU Lyon), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Mutant MESH: Protein Structure Secondary 030204 cardiovascular system & hematology Epitope Protein Structure Secondary law.invention MESH: Protein Structure Tertiary 0302 clinical medicine MESH: Structure-Activity Relationship law MESH: Chlorocebus aethiops hemic and lymphatic diseases Chlorocebus aethiops MESH: Animals MESH: von Willebrand Factor C2 domain Alanine biology [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] Chemistry Hematology MESH: Factor VIII MESH: COS Cells MESH: Mutagenesis Site-Directed Biochemistry MESH: Blood Coagulation COS Cells Recombinant DNA Blood Coagulation Tests Protein Binding MESH: Mutation MESH: Alanine In silico Cofactor 03 medical and health sciences Structure-Activity Relationship Von Willebrand factor von Willebrand Factor Animals Humans MESH: Protein Binding Blood Coagulation Factor VIII MESH: Humans MESH: Blood Coagulation Tests Protein Structure Tertiary 030104 developmental biology Mutation biology.protein Mutagenesis Site-Directed |
| Zdroj: | Thrombosis and Haemostasis Thrombosis and Haemostasis, 2017, 106 (07), pp.121-131. ⟨10.1160/TH10-09-0572⟩ Thrombosis and Haemostasis, Schattauer, 2017, 106 (07), pp.121-131. ⟨10.1160/TH10-09-0572⟩ |
| ISSN: | 0340-6245 |
| Popis: | SummaryThe factor VIII (FVIII) is a cofactor of the coagulation cascade. The FVIII C2 domain is a critical domain that participates in the interactions with the von Willebrand factor and the phospholipidic surfaces. To assess the importance of each residue of this domain in the maintenance of the structure and the function of FVIII, a number (n=139) of mutants were generated by substituting the original residues, from Ser2173 to Gly2325, by an alanine. Mutants were built within a complete B domain- deleted FVIII and expressed in COS-1 cells. Mutant antigen levels and procoagulant activities were measured. Two in silico analyses, a sliding average procedure and an analysis of the mutation energy cost were conducted in parallel on the FVIII structure. Both results were in agreement with the functional data, and illustrated the benefit of using such strategies prior to targeting specific residues in the aim of generating active recombinant molecules. The functional assays identify the residues that are important to maintaining the structure of the C2 domain, mainly those forming β-sheet, and those that can afford substitution, establishing a detailed functional relation with the available crystallographic data. This study provided a comprehensive functional mapping of the FVIII C2 domain and discussed the implication of specific residues in respect to the maintenance in the activity and structure stability, the efficiency in secretion, the binding to phospholipids and the formation of epitope. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|