Relating surfactant properties to activity and solubilization of the human adenosine A3 receptor
Relating surfactant properties to activity and solubilization of the human adenosine A3 receptor
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DOI：
10.1529/biophysj.104.051417
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发表时间：
2005-07-01
影响因子：
3.4
通讯作者：
Robinson, CR
中科院分区：
文献类型：
--
作者：
Berger, BW;García, RY;Robinson, CR
研究了不同表面活性剂对人腺苷A(3)受体（A(3)）活性和稳定性的影响。以每毫克膜蛋白44 pmol功能受体的浓度稳定转染HEK293细胞表达该受体，并使用50多种不同的非离子表面活性剂纯化。表面活性剂从膜上去除A(3)的能力与表面活性剂相对于其他膜蛋白选择性去除A(3)的能力之间存在很强的相关性。纯化后的A(3)活性也与所用表面活性剂的选择性密切相关。改变表面活性剂的效果比在纯化方案中加入A(3)配体的效果强得多。值得注意的是，所有具有高效率、选择性和活性的表面活性剂都落在亲水和亲脂平衡值的狭窄范围内。这种效应可能反映了表面活性剂在疏水跨膜界面上有效堆积的能力。这些发现强调了为特定膜蛋白鉴定合适的表面活性剂的重要性，并为开发快速、有效和系统的方法来促进膜蛋白纯化提供了希望。
The effects of various surfactants on the activity and stability of the human adenosine A(3) receptor (A(3)) were investigated. The receptor was expressed using stably transfected HEK293 cells at a concentration of 44 pmol functional receptor per milligram membrane protein and purified using over 50 different nonionic surfactants. A strong correlation was observed between a surfactant's ability to remove A(3) from the membrane and the ability of the surfactant to remove A(3) selectively relative to other membrane proteins. The activity of A(3) once purified also correlates well with the selectivity of the surfactant used. The effects of varying the surfactant were much stronger than those achieved by including A(3) ligands in the purification scheme. Notably, all surfactants that gave high efficiency, selectivity and activity fall within a narrow range of hydrophile-lipophile balance values. This effect may reflect the ability of the surfactant to pack effectively at the hydrophobic transmembrane interface. These findings emphasize the importance of identifying appropriate surfactants for a particular membrane protein, and offer promise for the development of rapid, efficient, and systematic methods to facilitate membrane protein purification.