Interhelical Packing in Detergent Micelles

Interhelical Packing in Detergent Micelles
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洗涤剂胶束中的螺旋间堆积

DOI:
10.1074/jbc.m110264200
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发表时间:
2002
期刊:
The Journal of Biological Chemistry
影响因子:
--
通讯作者:
C. Deber
C. Deber
中科院分区:
--
文献类型:
--
作者:
A. Therien;C. Deber

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利用由囊性纤维化跨膜电导调节器(CFTR)的相邻跨膜片段3和4组成的螺旋-环-螺旋结构,我们描述了一个研究多发性膜蛋白分子内螺旋-螺旋相互作用的系统。通过测量作为螺旋-螺旋接近度决定因素的Py准分子带强度,我们发现螺旋在洗涤剂胶束中保持三级接触。值得注意的是,胶束洗涤剂的性质可以改变这些接触的稳定性,全氟辛酸盐高度支持,溶血磷脂酰胆碱和溶血磷脂酰甘油略有耐受性,而十二烷基硫酸钠在很大程度上不能容忍这种相互作用。这一结构被进一步用于研究胶束洗涤剂的酰基链长在调节螺旋间堆积中的作用;具有9个碳的酰基链的洗涤剂显示最大程度的螺旋堆积。这些结果提供了关于脂类在膜蛋白折叠和构象上的作用的重要信息,并证明了基于芘的系统在研究支配螺旋间堆积的力方面的有效性。
Using a helix-loop-helix construct consisting of the adjacent transmembrane segments 3 and 4 of the cystic fibrosis transmembrane conductance regulator (CFTR) labeled with pyrene at both N and C termini, we describe a system for the study of intramolecular helix-helix interactions within a polytopic membrane protein. Through measurement of pyrene excimer band intensity as a determinant of helix-helix proximity, we show that the helices retain tertiary contacts in detergent micelles. Notably, the nature of the micellar detergent can alter the stability of these contacts, with perfluorooctanoate highly supportive, lysophosphatidylcholine and lysophosphatidylglycerol somewhat less tolerant, and SDS largely intolerant of such interactions. This construct is further employed to study the role of the acyl chain length of micellar detergents in modulating interhelical packing; detergents having acyl chains of 9 carbons display the greatest extent of helical packing. These results provide important information regarding the role of lipids on membrane protein folding and conformation as well as demonstrate the usefulness of a pyrene-based system in studying the forces that govern interhelical packing.
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