Giusti, Fabrice Kessler, Pascal Hansen, Randi Westh A. Della Pia, Eduardo Bon, Christel Le Mourier, Gilles Popot, Jean-Luc Martinez, Karen L. Zoonens, Manuela Synthesis of a Polyhistidine-bearing Amphipol and its Use for Immobilizing Membrane Proteins Amphipols (APols) are short amphipathic polymers that stabilize membrane proteins (MPs) in aqueous solutions. In the present study, A8–35, a polyacrylate-based APol, was grafted with hexahistidine tags (His<sub>6</sub>-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 His<sub>6</sub>-tag fused at one extremity of a MP, the presence of several His<sub>6</sub>-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. SPR;surface plasmon resonance;hexahistidine tags;amphipathic polymers;metal affinity chromatography;membrane proteins;Immobilizing Membrane ProteinsAmphipols;transmembrane domain;APol belt;IMAC resin;novel functionalized APol;MP increases;immobilize MPs;target protein 2015-12-14
    https://acs.figshare.com/articles/journal_contribution/Synthesis_of_a_Polyhistidine_bearing_Amphipol_and_its_Use_for_Immobilizing_Membrane_Proteins/2099824
10.1021/acs.biomac.5b01010.s001