Specific binding of integrin alphaIIbbeta3 to RGD peptide immobilized on a nitrilotriacetic acid chip: a surface plasmon resonance study |
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Authors: | Lu Y-J Zhang F Sui S-F |
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Affiliation: | (1) Department of Biological Sciences and Biotechnology, State Key Laboratory of Biomembranes, Tsinghua University, Beijing, 100084, P. R. China |
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Abstract: | 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--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. |
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Keywords: | nitrilotriacetic acid surface plasmon resonance RGD integrin /content/k602u87407h8x572/xxlarge945.gif" alt=" agr" align=" BASELINE" BORDER=" 0" >IIb /content/k602u87407h8x572/xxlarge946.gif" alt=" beta" align=" MIDDLE" BORDER=" 0" >3 His-tagged peptide |
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