Kinet J P, Alcaraz G, Leonard A, Wank S, Metzger H
Biochemistry. 1985 Jul 16;24(15):4117-24. doi: 10.1021/bi00336a046.
We previously showed that, in the absence of phospholipids, exposure of the tetrameric receptor for immunoglobulin E to mild detergents dissociates the intact beta chain and two gamma chains from the alpha chains. Having developed a practical method for assaying the dissociation, we have now explored a variety of different detergents, detergent concentrations, temperatures, times, salts, pHs, and other factors that influence the detergent-induced dissociation. Our findings should be useful for optimizing the stability of the receptor and for future studies on recombination of the subunits. The data suggest the following: (1) The critical perturbant is micellar detergent. (2) Unlike solubilization of membranes, where a molar ratio of micellar detergent:lipid of 2 is adequate, dissociation of the receptor is incomplete even at molar ratios of micellar detergent:receptor of greater than 10(5) and may be limited by a reversible component. (3) Detergents that are best for solubilizing membranes are also best for dissociating the receptors. (4) The latter observation and other data implicate bound lipid as stabilizing the receptor. Our findings may be applicable to the study of interactions between membrane proteins in general.
我们先前表明，在没有磷脂的情况下，将四聚体免疫球蛋白E受体暴露于温和的去污剂中会使完整的β链和两条γ链与α链分离。在开发出一种检测这种解离的实用方法后，我们现在研究了多种不同的去污剂、去污剂浓度、温度、时间、盐、pH值以及其他影响去污剂诱导解离的因素。我们的研究结果对于优化受体的稳定性以及未来亚基重组的研究应该是有用的。数据表明如下：（1）关键的扰动因素是胶束去污剂。（2）与膜的溶解不同，胶束去污剂与脂质的摩尔比为2就足以溶解膜，但即使胶束去污剂与受体的摩尔比大于10⁵，受体的解离也不完全，并且可能受到可逆成分的限制。（3）最适合溶解膜的去污剂也最适合使受体解离。（4）后一观察结果和其他数据表明结合的脂质可稳定受体。我们的研究结果可能普遍适用于膜蛋白之间相互作用的研究。
