Effects of solubilization on the structure and function of the sensory rhodopsin II/transducer complex
Effects of solubilization on the structure and function of the sensory rhodopsin II/transducer complex
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DOI：
10.1016/j.jmb.2005.12.015
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发表时间：
2006-03-10
影响因子：
5.6
通讯作者：
Engelhard, M
中科院分区：
文献类型：
--
作者：
Klare, JP;Bordignon, E;Engelhard, M
脂蛋白相互作用在膜整体蛋白的结构和生理活性中起着至关重要的作用。然而，目前的膜蛋白纯化技术需要从膜上提取到洗涤剂胶束中。此外，根据实验方案，膜蛋白的大部分可用数据都是使用洗涤剂增溶的样品获得的。因此，膜蛋白的稳定增溶是生物技术以及生物化学和结构生物学中的一个重要问题。了解增溶作用对特定蛋白质的结构和功能性质的影响对于评价和解释实验结果是非常重要的。在这项研究中，比较了古细菌光感受器/转导复合体(NpSRII/NPHtrII)在洗涤剂溶解和脂类重组状态下的结构和动力学数据。激光闪光光解、荧光光谱和电子顺磁共振光谱数据表明，增溶作用对受体蛋白的光循环动力学和转导蛋白的结构有相当大的影响。尤其是蛋白质-膜近端区域和蛋白质-蛋白质界面区域对非天然条件敏感。这些数据表明，从溶解的膜蛋白或膜蛋白复合体中获得的生化和结构信息的相关性必须仔细评估。(C)2006爱思唯尔有限公司。保留所有权利。
Lipid-protein interactions are known to play a crucial role in structure and physiological activity of integral membrane proteins. However, current technology for membrane protein purification necessitates extraction from the membrane into detergent micelles. Also, due to experimental protocols, most of the data available for membrane proteins is obtained using detergent-solubilized samples. Stable solubilization of membrane proteins is therefore an important issue in biotechnology as well as in biochemistry and structural biology. An understanding of solubilization effects on structural and functional properties of specific proteins is of utmost relevance for the evaluation and interpretation of experimental results. In this study, a comparison of structural and kinetic data obtained for the archaebacterial photoreceptor/transducer complex from Natronomonas pharaonis (NpSRII/NpHtrII) in detergent-solubilized and lipid-reconstituted states is presented. Laser flash photolysis, fluorescence spectroscopy, and electron paramagnetic resonance spectroscopy data reveal considerable influence of solubilization on the photocycle kinetics of the receptor protein and on the structure of the transducer protein. Especially the protein-membrane proximal region and the protein-protein interfacial domains are sensitive towards non-native conditions. These data demonstrate that relevance of biochemical and structural information obtained from solubilized membrane proteins or membrane protein complexes has to be evaluated carefully. (c) 2006 Elsevier Ltd. All rights reserved.