Synthesis of a Polyhistidine-bearing Amphipol and its Use for Immobilizing Membrane Proteins

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• 作者：Fabrice Giusti ; Pascal Kessler ; Randi Westh Hansen ; Eduardo A. Della Pia ; Christel Le Bon ; Gilles Mourier ; Jean-Luc Popot ; Karen L. Martinez ; Manuela Zoonens
• 刊名：Biomacromolecules
• 出版年：2015
• 出版时间：December 14, 2015
• 年：2015
• 卷：16
• 期：12
• 页码：3751-3761
• 全文大小：608K
• ISSN：1526-4602

文摘

Amphipols (APols) are short amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions. In the present study, A8鈥?5, a polyacrylate-based APol, was grafted with hexahistidine tags (His₆-tags). The synthesis and characterization of this novel functionalized APol, named HistAPol, are described. Its ability to immobilize MPs on nickel ion-bearing surfaces was tested using two complementary methods, immobilized metal affinity chromatography (IMAC) and surface plasmon resonance (SPR). Compared to a single His₆-tag fused at one extremity of a MP, the presence of several His₆-tags carried by the APol belt surrounding the transmembrane domain of a MP increases remarkably the affinity of the protein/APol complex for nickel ion-bearing SPR chips, whereas it does not show such a strong effect on an IMAC resin. HistAPol-mediated immobilization, which allows reversibility of the interaction and easy regeneration of the supports and dispenses with any genetic modification of the target protein, provides a novel, promising tool for attaching MPs onto solid supports while stabilizing them.