The interaction of trimethylamine dehydrogenase and electron-transferring flavoprotein.
The interaction between the physiological electron transfer partners trimethylamine dehydrogenase (TMADH) and electron-transferring flavoprotein (ETF) from Methylophilus methylotrophus has been examined with particular regard to the proposal that the former protein "imprints" a conformational change on the latter. The results indicate that the absorbance change previously attributed to changes in the environment of the FAD of ETF upon binding to TMADH is instead caused by electron transfer from partially reduced, as-isolated TMADH to ETF. Prior treatment of the as-isolated enzyme with the oxidant ferricenium essentially abolishes the observed spectral change. Further, when the semiquinone form of ETF is used instead of the oxidized form, the mirror image of the spectral change seen with as-isolated TMADH and oxidized ETF is observed. This is attributable to a small amount of electron transfer in the reverse of the physiological direction. Kinetic determination of the dissociation constant and limiting rate constant for electron transfer within the complex of (reduced) TMADH with (oxidized) ETF is reconfirmed and discussed in the context of a recently proposed model for the interaction between the two proteins that involves "structural imprinting" of ETF.
Shi
Weiwei
W
Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus, Ohio 43210-1218, USA.
Mersfelder
John
J
Hille
Russ
R
D016227:Benzoquinones; D004227:Dithionite / Q000737:chemistry; D044927:Electron-Transferring Flavoproteins / Q000737:chemistry* / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005182:Flavin-Adenine Dinucleotide / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D020630:Methylophilus methylotrophus / Q000737:chemistry; D010084:Oxidation-Reduction; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D010659:Phenylhydrazines / Q000494:pharmacology; D011487:Protein Conformation; D013053:Spectrophotometry
D016227:Benzoquinones; D044927:Electron-Transferring Flavoproteins; D010659:Phenylhydrazines; C030299:phenylhydrazine; D005182:Flavin-Adenine Dinucleotide; D004227:Dithionite; C025232:semiquinone radicals; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase