Kinetics of Membrane Protein-Detergent Interactions Depend on Protein Electrostatics.
Kinetics of Membrane Protein-Detergent Interactions Depend on Protein Electrostatics.
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膜蛋白-去污剂相互作用的动力学取决于蛋白质静电。
DOI：
10.1021/acs.jpcb.8b07889
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Movileanu,Liviu
中科院分区：
文献类型：
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作者：
Wolfe,AaronJ;Gugel,JackF;Chen,Min;Movileanu,Liviu
膜蛋白与洗涤剂胶束的相互作用代表了结构和化学生物学中具有实际意义的基本过程。蛋白质-去污剂复合物（PDC）相互作用动力学的定量评估一直受到膜蛋白和增溶去污剂在水相中的复杂行为的挑战。在这里，我们显示了含麦芽糖苷的洗涤剂从β-桶膜蛋白解吸的动力学读数。使用稳态荧光偏振（FP）各向异性测量，我们记录了实时的，特定的签名的PDC相互作用。这些测量的结果被用来推断模型依赖的速率常数的协会和解离的蛋白质胶束。值得注意的是，PDC相互作用的动力学取决于总的蛋白质电荷，尽管洗涤剂单体的非离子性质。在未来，这种方法可能被用于高通量筛选的动力学指纹的不同膜蛋白稳定在胶束中，含有各种洗涤剂的混合物。
Interactions of a membrane protein with a detergent micelle represent a fundamental process with practical implications in structural and chemical biology. Quantitative assessment of the kinetics of protein–detergent complex (PDC) interactions has always been challenged by complicated behavior of both membrane proteins and solubilizing detergents in aqueous phase. Here, we show the kinetic reads of the desorption of maltoside-containing detergents from β-barrel membrane proteins. Using steady-state fluorescence polarization (FP) anisotropy measurements, we recorded real-time, specific signatures of the PDC interactions. The results of these measurements were used to infer the model-dependent rate constants of association and dissociation of the proteomicelles. Remarkably, the kinetics of the PDC interactions depend on the overall protein charge despite the nonionic nature of the detergent monomers. In the future, this approach might be employed for high-throughput screening of kinetic fingerprints of different membrane proteins stabilized in micelles that contain mixtures of various detergents.