Steroid-based facial amphiphiles for stabilization and crystallization of membrane proteins
Steroid-based facial amphiphiles for stabilization and crystallization of membrane proteins
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DOI：
10.1073/pnas.1221442110
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发表时间：
2013-03-26
影响因子：
11.1
通讯作者：
Zhang, Qinghai
中科院分区：
文献类型：
--
作者：
Lee, Sung Chang;Bennett, Brad C.;Zhang, Qinghai
两亲体选择是膜蛋白（MPs）结构研究的关键步骤。我们已经开发了一个基于类固醇的面部两亲化合物（FAs）家族，它在结构上与传统的洗涤剂和以前开发的FAs不同。独特的FAs稳定MPs并形成相对较小的蛋白质-洗涤剂复合物（PDCs），这一特性被认为有利于MP的结晶。我们试图结晶几种属于不同蛋白家族的MPs，包括人类间隙连接通道蛋白连接蛋白26、ATP结合盒转运蛋白MsbA、7跨膜G蛋白偶联受体样细菌视紫红质和细胞色素p450（外周MPs）。单独使用FAs或与其他洗涤剂或脂类混合，我们获得了适合x射线晶体学分析的上述蛋白质的3D晶体。与传统的洗涤剂相比，FAs提高了聚羧酸的结晶性，这可以归因于几个特性，包括提高了蛋白质的稳定性，形成了小聚羧酸，降低了聚羧酸的表面柔韧性，以及调节晶格接触的潜力。
Amphiphile selection is a critical step for structural studies of membrane proteins (MPs). We have developed a family of steroid-based facial amphiphiles (FAs) that are structurally distinct from conventional detergents and previously developed FAs. The unique FAs stabilize MPs and form relatively small protein-detergent complexes (PDCs), a property considered favorable for MP crystallization. We attempted to crystallize several MPs belonging to different protein families, including the human gap junction channel protein connexin 26, the ATP binding cassette transporter MsbA, the seven-transmembrane G protein-coupled receptor-like bacteriorhodopsin, and cytochrome P450s (peripheral MPs). Using FAs alone or mixed with other detergents or lipids, we obtained 3D crystals of the above proteins suitable for X-ray crystallographic analysis. The fact that FAs enhance MP crystallizability compared with traditional detergents can be attributed to several properties, including increased protein stability, formation of small PDCs, decreased PDC surface flexibility, and potential to mediate crystal lattice contacts.