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

TY - JOUR

T1 - Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins

AU - Giusti, Fabrice

AU - Kessler, Pascal

AU - Hansen, Randi Westh

AU - Della Pia, Eduardo Antonio

AU - Le Bon, Christel

AU - Mourier, Gilles

AU - Popot, Jean-Luc

AU - Martinez, Karen Laurence

AU - Zoonens, Manuela

PY - 2015/10/22

Y1 - 2015/10/22

N2 - 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.

AB - 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.

U2 - 10.1021/acs.biomac.5b01010

DO - 10.1021/acs.biomac.5b01010

M3 - Journal article

C2 - 26492302

SN - 1525-7797

VL - 16

SP - 3751

EP - 3761

JO - Biomacromolecules

JF - Biomacromolecules

IS - 12

ER -