Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins

Fabrice Giusti, Pascal Kessler, Randi Westh Hansen, Eduardo Antonio Della Pia, Christel Le Bon, Gilles Mourier, Jean-Luc Popot, Karen Laurence Martinez, Manuela Zoonens

12 Citationer (Scopus)

Abstract

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 (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.

OriginalsprogEngelsk
TidsskriftBiomacromolecules
Vol/bind16
Udgave nummer12
Sider (fra-til)3751-3761
Antal sider11
ISSN1525-7797
DOI
StatusUdgivet - 22 okt. 2015

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