title: Biomimetic environment to study E. coli complex I through surface-enhanced IR absorption spectroscopy.
abstract: In this study complex I was immobilized in a biomimetic environment on a gold layer deposited on an ATR-crystal in order to functionally probe the enzyme against substrates and inhibitors via surface-enhanced IR absorption spectroscopy (SEIRAS) and cyclic voltammetry (CV). To achieve this immobilization, two methods based on the generation of a high affinity self-assembled monolayer (SAM) were probed. The first made use of the affinity of Ni-NTA toward a hexahistidine tag that was genetically engineered onto complex I and the second exploited the affinity of the enzyme toward its natural substrate NADH. Experiments were also performed with complex I reconstituted in lipids. Both approaches have been found to be successful, and electrochemically induced IR difference spectra of complex I were obtained.
authors.affiliation: Laboratoire de bioelectrochimie et spectroscopie, UMR 7140, Chimie de la Matière Complexe, Université de Strasbourg-CNRS , Strasbourg 67000, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Kriegel
authors.forename: Taro
authors.initials: T
authors.lastname: Uchida
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Osawa
authors.forename: Thorsten
authors.initials: T
authors.lastname: Friedrich
authors.forename: Petra
authors.initials: P
authors.lastname: Hellwig
mesh_terms: D042967:Electron Transport Complex I / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D006046:Gold / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry; D018716:Molecular Mimicry; D009243:NAD / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D004800:Enzymes, Immobilized; D029968:Escherichia coli Proteins; D008051:Lipid Bilayers; D009243:NAD; D006046:Gold; D042967:Electron Transport Complex I