Structure and Ca²⁺-binding properties of the tandem C₂ domains of E-Syt2.

Structure and Ca²⁺-binding properties of the tandem C₂ domains of E-Syt2.
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DOI:
10.1016/j.str.2013.11.011
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发表时间:
2014-02-04
期刊:
影响因子:
5.7
通讯作者:
Rizo, Josep
Rizo, Josep
中科院分区:
生物学2区
文献类型:
--
作者:
Xu, Junjie;Bacaj, Taulant;Zhou, Amy;Tomchick, Diana R.;Suedhof, Thomas C.;Rizo, Josep

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内质网和质膜之间的接触涉及哺乳动物中的延伸突触结合蛋白(E-Syts)或酵母中的三白蛋白(具有多个C2结构域的蛋白质)。E-Syt 2的串联C2结构域之一被预测为结合Ca 2+,但没有Ca 2+依赖性功能已归因于该蛋白质。我们已经确定了E-Syt 2的串联C2结构域的晶体结构的情况下和存在的Ca 2+,并分析其Ca 2+结合的性质,通过NMR光谱。我们的数据揭示了一个意想不到的V形结构与刚性取向之间的两个C2域,是没有实质性改变的Ca 2+。E-Syt 2 C2 A结构域结合多达四个Ca 2+离子,而C2B结构域不结合Ca 2+。这些结果表明,E-Syt 2执行一个尚未确定的Ca 2+依赖性功能,通过其C2 A域,并揭示E-Syts和突触结合蛋白的串联C2域的属性之间的根本差异。
Contacts between the endoplasmic reticulum and the plasma membrane involve extended synaptotagmins (E-Syts) in mammals or tricalbins in yeast, proteins with multiple C2 domains. One of the tandem C2 domains of E-Syt2 is predicted to bind Ca2+, but no Ca2+-dependent function has been attributed to this protein. We have determined the crystal structures of the tandem C2 domains of E-Syt2 in the absence and presence of Ca2+, and analyzed their Ca2+-binding properties by NMR spectroscopy. Our data reveal an unexpected V-shaped structure with a rigid orientation between the two C2 domains that is not substantially altered by Ca2+. The E-Syt2 C2A domain binds up to four Ca2+ ions, whereas the C2B domain does not bind Ca2+. These results suggest that E-Syt2 performs an as yet unidentified Ca2+-dependent function through its C2A domain, and uncover fundamental differences between the properties of the tandem C2 domains of E-Syts and synaptotagmins.
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