Electrophoretic mobility shift in native gels indicates calcium-dependent structural changes of neuronal calcium sensor proteins

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• 作者：Jeffrey Viviano^a ; Anuradha Krishnan^a ; Hao Wu^a ; Venkat Venkataraman^a ; ^b ; ^{vvenkat2007@gmail.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Neuronal calcium sensor proteins ; Electrophoresis ; Native gels ; Calcium
• 刊名：Analytical Biochemistry
• 出版年：2016
• 出版时间：1 February 2016
• 年：2016
• 卷：494
• 期：Complete
• 页码：93-100
• 全文大小：971 K

文摘

In proteins of the neuronal calcium sensor (NCS) family, changes in structure as well as function are brought about by the binding of calcium. In this article, we demonstrate that these structural changes, solely due to calcium binding, can be assessed through electrophoresis in native gels. The results demonstrate that the NCS proteins undergo ligand-dependent conformational changes that are detectable in native gels as a gradual decrease in mobility with increasing calcium but not other tested divalent cations such as magnesium, strontium, and barium. Surprisingly, such a gradual change over the entire tested range is exhibited only by the NCS proteins but not by other tested calcium-binding proteins such as calmodulin and S100B, indicating that the change in mobility may be linked to a unique NCS family feature—the calcium–myristoyl switch. Even within the NCS family, the changes in mobility are characteristic of the protein, indicating that the technique is sensitive to the individual features of the protein. Thus, electrophoretic mobility on native gels provides a simple and elegant method to investigate calcium (small ligand)-induced structural changes at least in the superfamily of NCS proteins.