On-bead purification and nanodisc reconstitution of human chemokine receptor complexes for structural and biophysical studies

人类趋化因子受体复合物的珠上纯化和纳米盘重建,用于结构和生物物理研究

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作者:Siyi Gu, Mian Huang, Tracy M Handel

Abstract

Chemokine receptors, a subfamily of G-protein-coupled receptors (GPCRs), are responsible for cell migration during physiological processes as well as in diseases like inflammation and cancers. Here, we present a protocol for solubilizing, purifying, and reconstituting complexes of chemokine receptors with their ligands in "nanodiscs," soluble lipid bilayers that mimic the native environment of membrane receptors. The protocol yields chemokine receptor complexes with sufficient purity and yield for structural and biophysical studies and should be applicable to other GPCRs.

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