人类srGAP2蛋白反向F-BAR结构域的纯化、结晶及初步X射线分析
Purification, crystallization and preliminary X-ray analysis of the inverse F-BAR domain of the human srGAP2 protein.
作者信息
Wang Hongpeng, Zhang Yan, Zhang Zhenyi, Jin Wei Lin, Wu Geng
机构信息
State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
出版信息
Acta Crystallogr F Struct Biol Commun. 2014 Jan;70(Pt 1):123-6. doi: 10.1107/S2053230X13033712. Epub 2013 Dec 24.
Bin-Amphiphysin-Rvs (BAR) domain proteins play essential roles in diverse cellular processes by inducing membrane invaginations or membrane protrusions. Among the BAR superfamily, the `classical' BAR and Fes/CIP4 homology BAR (F-BAR) subfamilies of proteins usually promote membrane invaginations, whereas the inverse BAR (I-BAR) subfamily generally incur membrane protrusions. Despite possessing an N-terminal F-BAR domain, the srGAP2 protein regulates neurite outgrowth and neuronal migration by causing membrane protrusions reminiscent of the activity of I-BAR domain proteins. In this study, the inverse F-BAR (IF-BAR) domain of human srGAP2 was overexpressed, purified and crystallized. The crystals of the srGAP2 IF-BAR domain protein diffracted to 3.50 Å resolution and belonged to space group P2(1). These results will facilitate further structural determination of the srGAP2 IF-BAR domain and the ultimate elucidation of its peculiar behaviour of inducing membrane protrusions rather than membrane invaginations.
Bin-发动蛋白-Rvs（BAR）结构域蛋白通过诱导膜内陷或膜突出在多种细胞过程中发挥重要作用。在BAR超家族中，“经典”BAR和Fes/CIP4同源性BAR（F-BAR）亚家族蛋白通常促进膜内陷，而反向BAR（I-BAR）亚家族一般引起膜突出。尽管srGAP2蛋白具有一个N端F-BAR结构域，但它通过引起类似于I-BAR结构域蛋白活性的膜突出，来调节神经突生长和神经元迁移。在本研究中，人srGAP2的反向F-BAR（IF-BAR）结构域被过量表达、纯化并结晶。srGAP2 IF-BAR结构域蛋白的晶体衍射分辨率达到3.50 Å，属于P2(1)空间群。这些结果将有助于进一步确定srGAP2 IF-BAR结构域的结构，并最终阐明其诱导膜突出而非膜内陷的特殊行为。
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