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首页> 外文期刊>The FEBS journal >The second C2-domain of copine-2, copine-6 and copine-7 is responsible for their calcium-dependent membrane association
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The second C2-domain of copine-2, copine-6 and copine-7 is responsible for their calcium-dependent membrane association

机译：copine-2，copine-6和copine-7的第二个C2结构域负责其钙依赖性膜结合

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摘要

The copine family of proteins contains nine members with a similar domain structure, namely two N-terminal C2-domains (C2A and C2B) and a C-terminal A-domain. The former are thought to be responsible for binding to the inner face of the plasma membrane following increases in intracellular calcium levels, whereas the A-domain has been suggested to be a protein-binding structure. In this study, we examined the effects of mutagenesis of selected residues in the linker area between the C2-domains and the A-domain, and mutagenesis of the aspartates of the C2-domains, which are predicted to bind calcium and promote membrane association of the copines. We found that Lys282-Lys284 of the linker area are important for the folding of the intact protein. We showed that substitution with asparagine, single or multiple, of the aspartates in the C2A-domain had no effect on the calcium-mediated membrane association of copine-2, copine-6, or copine-7. Similar mutagenesis of a single residue in the C2B-domain of copine-6 (but not copine-2 and copine-7) was sufficient to eliminate its calcium-mediated membrane binding, and simultaneous substitution of all four of the asparagines in the C2B-domain resulted in constitutive membrane association of copine-2, copine-6 and copine-7 with the plasma membrane. These data show that the C2B-domains of copine-2, copine-6 and copine-7 are the domains responsible for the protein calcium-dependent membrane association.
机译：蜂胶蛋白家族包含具有相似结构域结构的九个成员，即两个N端C2域（C2A和C2B）和一个C端A域。认为前者负责细胞内钙水平增加后与质膜内表面的结合，而A结构域被认为是一种蛋白质结合结构。在这项研究中，我们检查了C2域和A域之间的接头区域中选定残基的诱变作用以及C2域的天冬氨酸的诱变作用，这些突变预计会结合钙并促进膜的缔合。菲律宾人。我们发现，接头区域的Lys282-Lys284对于完整蛋白的折叠很重要。我们表明，在C2A域中用天冬氨酸（单个或多个）替换天冬酰胺对copine-2，copine-6或copine-7的钙介导的膜缔合没有影响。对copine-6的C2B域中的单个残基（但不是copine-2和copine-7）进行类似的诱变足以消除其钙介导的膜结合，并同时取代C2B-中的所有四个天冬酰胺。结构域导致copine-2，copine-6和copine-7与质膜的本构膜缔合。这些数据表明copine-2，copine-6和copine-7的C2B结构域是负责蛋白质钙依赖性膜结合的结构域。

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