Bead-linked Proteoliposomes: A Reconstitution Method for NMR Analyses of Membrane Protein-Ligand Interactions

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• 作者：Mariko Yokogawa ; Koh Takeuchi ; and Ichio Shimada
• 刊名：Journal of the American Chemical Society
• 出版年：2005
• 出版时间：August 31, 2005
• 年：2005
• 卷：127
• 期：34
• 页码：12021 - 12027
• 全文大小：247K
• 年卷期：v.127,no.34(August 31, 2005)
• ISSN：1520-5126

文摘

Structural information about the interactions between membrane proteins and their ligandsprovides insights into the membrane protein functions. A variety of surfactants have been used for structuralanalyses of membrane proteins, and in some cases, they yielded successful results. However, the use ofsurfactants frequently increases the conformational instability of membrane proteins and distorts their normalfunction. Here, we propose a new strategy of membrane protein reconstitution into lipid bilayers on affinitybeads, which maintains the native conformation and function of the protein for NMR studies. Thereconstituted membrane proteins are suitable for NMR analyses of interactions, by using the transferredcross-saturation method. The strategy was successfully applied to the interaction between a potassiumion channel, KcsA, and a pore-blocker, agitoxin2 (AgTx2). This strategy would be useful for analyzing theinteractions between various membrane proteins and their ligands.