Specific binding of integrin alphaIIbbeta3 to RGD peptide immobilized on a nitrilotriacetic acid chip: a surface plasmon resonance study.
Nitrilotriacetic acid has been routinely used in protein purification for its high affinity for His-tagged protein in the presence of Ni2+. Here we reported a type of nitrilotriacetic acid chip (NTA-chip) prepared by transferring NTA-DOGS containing a lipid monolayer to a 50 nm thick gold layer deposited on a glass slide. The surface binding ability of His-tagged protein and regeneration of NTA chip were characterized using a synthetic polypeptide P1 (His-His-His-His-His-His-epsilon-aminohexanoic-Gly-Gly-Arg-Gly-Asp-Ser). The effect of divalent cations on integrin binding affinity for RGD ligand was investigated after P1 had been immobilized onto the sensor chip. The results show that the NTA-chip is a useful tool to immobilize His-tagged protein on the chip surface, and can provide a functional orientation for further investigation. The results also show that removing of Ca2+ bound on low affinity sites or adding of Mn2+ can increase the binding ability of integrin.
Lu
Y-J
YJ
Department of Biological Sciences and Biotechnology, State Key Laboratory of Biomembranes, Tsinghua University, Beijing 100084, P. R. China.
Zhang
F
F
Sui
S-F
SF
D008239:Lysine / Q000031:analogs & derivatives / Q000737:chemistry*; D009571:Nitrilotriacetic Acid / Q000737:chemistry*; D009829:Oleic Acids / Q000737:chemistry*; D009842:Oligopeptides / Q000378:metabolism*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry*; D011485:Protein Binding; D013379:Substrate Specificity; D013386:Succinates / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
C476561:N(alpha),N(alpha)-bis(carboxymethyl)-N(epsilon)-(1,2-dioleyl-sn-glycero-3-succinyl)-L-lysine; D009829:Oleic Acids; D009842:Oligopeptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D013386:Succinates; C047981:arginyl-glycyl-aspartic acid; D008239:Lysine; D009571:Nitrilotriacetic Acid