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  • Title: Biomimetic environment to study E. coli complex I through surface-enhanced IR absorption spectroscopy.
    Author: Kriegel S, Uchida T, Osawa M, Friedrich T, Hellwig P.
    Journal: Biochemistry; 2014 Oct 14; 53(40):6340-7. PubMed ID: 25225967.
    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.
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