Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins
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Synthesis of a polyhistidine-bearing amphipol and its use for immobilizing membrane proteins. / Giusti, Fabrice; Kessler, Pascal; Hansen, Randi Westh; Della Pia, Eduardo Antonio; Le Bon, Christel; Mourier, Gilles; Popot, Jean-Luc; Martinez, Karen Laurence; Zoonens, Manuela.
I: Biomacromolecules, Bind 16, Nr. 12, 2015, s. 3751-3761.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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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
Y1 - 2015
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
VL - 16
SP - 3751
EP - 3761
JO - Biomacromolecules
JF - Biomacromolecules
SN - 1525-7797
IS - 12
ER -
ID: 147934539