title: Formation of W(3)A(1) electron-transferring flavoprotein (ETF) hydroquinone in the trimethylamine dehydrogenase x ETF protein complex.
abstract: The electron-transferring flavoprotein (ETF) from Methylophilus methylotrophus (sp. W(3)A(1)) exhibits unusual oxidation-reduction properties and can only be reduced to the level of the semiquinone under most circumstances (including turnover with its physiological reductant, trimethylamine dehydrogenase (TMADH), or reaction with strong reducing reagents such as sodium dithionite). In the present study, we demonstrate that ETF can be reduced fully to its hydroquinone form both enzymatically and chemically when it is in complex with TMADH. Quantitative titration of the TMADH x ETF protein complex with sodium dithionite shows that a total of five electrons are taken up by the system, indicating that full reduction of ETF occurs within the complex. The results indicate that the oxidation-reduction properties of ETF are perturbed upon binding to TMADH, a conclusion further supported by the observation of a spectral change upon formation of the TMADH x ETF complex that is due to a change in the environment of the FAD of ETF. The results are discussed in the context of ETF undergoing a conformational change during formation of the TMADH x ETF electron transfer complex, which modulates the spectral and oxidation-reduction properties of ETF such that full reduction of the protein can take place.
authors.affiliation: Department of Medical Biochemistry, Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: M H
authors.initials: MH
authors.lastname: Jang
authors.forename: N S
authors.initials: NS
authors.lastname: Scrutton
authors.forename: R
authors.initials: R
authors.lastname: Hille
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D004227:Dithionite / Q000378:metabolism; D044927:Electron-Transferring Flavoproteins; D004583:Electrons; D005420:Flavoproteins / Q000737:chemistry* / Q000378:metabolism; D006873:Hydroquinones / Q000737:chemistry* / Q000378:metabolism; D020630:Methylophilus methylotrophus / Q000737:chemistry; D010084:Oxidation-Reduction; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry* / Q000378:metabolism; D010659:Phenylhydrazines / Q000494:pharmacology; D011485:Protein Binding; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D044927:Electron-Transferring Flavoproteins; D005420:Flavoproteins; D006873:Hydroquinones; D010659:Phenylhydrazines; C030299:phenylhydrazine; D004227:Dithionite; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase; C031927:hydroquinone