CHOBIMALT: A Cholesterol-Based Detergent
CHOBIMALT: A Cholesterol-Based Detergent
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DOI：
10.1021/bi101334j
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发表时间：
2010-11-09
期刊：
影响因子：
2.9
通讯作者：
Sanders, Charles R.
中科院分区：
文献类型：
--
作者：
Howell, Stanley C.;Mittal, Ritesh;Sanders, Charles R.
胆固醇及其琥珀酸半酯和硫酸盐衍生物被广泛应用于纯化膜蛋白的研究中，但即使在洗涤剂胶束存在的情况下，也很难在水溶液中溶解。其他胆固醇衍生物不会形成传统的胶束，从而产生粘性溶液。为了解决这些问题，一种以胆固醇为基础的洗涤剂CHOBIMALT被合成并进行了表征。在3-4微米以上的浓度下，CHOBIMALT形成胶束，不需要高温或声波破坏。扩散和荧光测量表明，CHOBIMALT胶束较大(210+/-30 kDa)。利用G蛋白偶联受体，人kappa阿片受体1型(HKOR1)，探索了溶解功能性膜蛋白的能力。虽然CHOBIMALT本身并不能有效地作为膜萃取的表面活性剂，但当CHOBIMALT被添加到经典洗涤剂胶束中时，可以观察到CHOBIMALT显著提高了增溶的hKOR1的热稳定性。
Cholesterol and its hemisuccinate and sulfate derivatives are widely used in studies of purified membrane proteins but are difficult to solubilize in aqueous solution, even in the presence of detergent micelles. Other cholesterol derivatives do not form conventional micelles and lead to viscous solutions. To address these problems, a cholesterol-based detergent, CHOBIMALT, has been synthesized and characterized. At concentrations above 3-4 mu M, CHOBIMALT forms micelles without the need for elevated temperatures or sonic disruption. Diffusion and fluorescence measurements indicated that CHOBIMALT micelles are large (210 +/- 30 kDa). The ability to solubilize a functional membrane protein was explored using it G-protein coupled receptor, the human kappa opioid receptor type 1 (hKOR1). While CHOBIMALT alone was not found to be effective as a surfactant for membrane extraction, when added to classical detergent micelles CHOBIMALT was observed to dramatically enhance the thermal stability of solubilized hKOR1.