TY - JOUR AU - Ataka, K. AU - Giess, F. AU - Knoll, W. AU - Naumann, R. AU - Haber-Pohlmeier, S. AU - Richter, B. AU - Heberle, J. TI - Oriented attachment and membrane reconstitution of His-tagged cytochrome c oxidase to a gold electrode: in-situ monitoring by Surface Enhanced Infrared Absorption Spectroscopy JO - Journal of the American Chemical Society VL - 126 SN - 0002-7863 CY - Washington, DC PB - American Chemical Society M1 - PreJuSER-43304 SP - 16199 - 16206 PY - 2004 N1 - Record converted from VDB: 12.11.2012 AB - A novel concept is introduced for the oriented incorporation of membrane proteins into solid supported lipid bilayers. Recombinant cytochrome c oxidase solubilized in detergent was immobilized on a chemically modified gold surface via the affinity of its histidine-tag to a nickel-chelating nitrilo-triacetic acid (NTA) surface. The oriented protein monolayer was reconstituted into the lipid environment by detergent substitution. The individual steps of the surface modification, including (1) chemical modification of the gold support, (2) adsorption of the protein, and (3) reconstitution of the lipid bilayer, were followed in situ by means of surface-enhanced infrared absorption spectroscopy (SEIRAS) and accompanied by normal-mode analysis. The high surface sensitivity of SEIRAS allows for the identification of each chemical reaction process within the monolayer at the molecular level. Finally, full functionality of the surface-tethered cytochrome c oxidase was demonstrated by cyclic voltammetry after binding of the natural electron donor cytochrome c. KW - Electrochemistry KW - Electron Transport Complex IV: chemistry KW - Enzymes, Immobilized: chemistry KW - Gold: chemistry KW - Histidine: chemistry KW - Lipid Bilayers: chemistry KW - Models, Molecular KW - Nitrilotriacetic Acid: chemistry KW - Recombinant Proteins: chemistry KW - Spectroscopy, Fourier Transform Infrared: methods KW - Surface Properties KW - Enzymes, Immobilized (NLM Chemicals) KW - Lipid Bilayers (NLM Chemicals) KW - Recombinant Proteins (NLM Chemicals) KW - Nitrilotriacetic Acid (NLM Chemicals) KW - Histidine (NLM Chemicals) KW - Gold (NLM Chemicals) KW - Electron Transport Complex IV (NLM Chemicals) KW - J (WoSType) LB - PUB:(DE-HGF)16 C6 - pmid:15584756 UR - <Go to ISI:>//WOS:000225697000055 DO - DOI:10.1021/ja045951h UR - https://juser.fz-juelich.de/record/43304 ER -