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High-throughput crystallographic screening device and method for crystalizing membrane proteins using a sub physiological resting membrane potential across a lipid matrix of variable composition

机译:高通量结晶学筛选设备和方法,利用跨组成可变的脂质基质的亚生理静息膜电位来结晶膜蛋白

摘要

The invention is a high-throughput voltage screening crystallographic device and methodology that uses multiple micro wells and electric circuits capable of assaying different crystallization condition for the same or different proteins of interest at the same of different voltages under a humidity and temperature controlled environment. The protein is solubilized in a lipid matrix similar to the lipid composition of the protein in the native environment to ensure stability of the protein during crystallization. The invention provides a system and method where the protein is transferred to a lipid matrix that holds a resting membrane potential, which reduces the degree of conformational freedom of the protein. The invention overcomes the majority of the difficulties associated with vapor diffusion techniques and essentially reconstitutes the protein in its native lipid environment under “cuasi” physiological conditions.
机译:本发明是使用多个微孔和电路的高通量电压筛选晶体学设备和方法,所述微孔和电路能够在湿度和温度受控的环境下以相同的不同电压测定相同或不同目的蛋白质的不同结晶条件。蛋白质溶解在类似于天然环境中蛋白质脂质组成的脂质基质中,以确保蛋白质在结晶过程中的稳定性。本发明提供了一种系统和方法,其中将蛋白质转移到具有静止膜电位的脂质基质中,这降低了蛋白质的构象自由度。本发明克服了与蒸气扩散技术有关的大多数困难,并在“易变”的生理条件下在其天然脂质环境中基本上重构了蛋白质。

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