The interaction of methanol dehydrogenase and its electron acceptor, cytochrome cL in methylotrophic bacteria.
The interactions of methanol dehydrogenase (MDH, EC1.1.99.8) with its specific electron acceptor cytochrome cL has been investigated in Methylobacterium extorquens and Methylophilus methylotrophus. The MDHs of these two very different methylotrophs have the same alpha 2 beta 2 structure; the interaction of these MDHs with their specific electron acceptor, cytochrome cL, has been studied using a novel assay system. Electrostatic reactions are involved in 'docking' of the two proteins. EDTA inhibits the reaction by a process involving neither metal chelation nor the 'docking' process. Chemical modification studies showed that the two proteins interact by a 'docking' process involving interactions of lysyl residues on MDH and carboxyl residues on cytochrome cL. When 'zero length', two stage cross-linking was done (with proteins from both bacteria), the alpha-subunits of MDH cross-linked with cytochrome cL by way of lysyl groups on MDH and carboxyl groups on the cytochrome. Tuna mitochondrial cytochrome c provided a model for cytochrome cH which is the electron acceptor for cytochrome cL in the 'methanol oxidase' electron transport chain. Tuna cytochrome c was shown to form crosslinked products with carboxyl-modified cytochrome cL. MDH and tuna cytochrome c competed for the same domain on cytochrome cL. It was concluded that MDH reacts with cytochrome cL by an electrostatic reaction which involves carboxyl groups on cytochrome cL and amino groups on the alpha-subunit of MDH. The same domain on cytochrome cL is involved in subsequent 'docking' with its electron acceptor.
Cox
J M
JM
SERC Molecular Recognition Centre, Biochemistry Department, University of Southampton, U.K.
Day
D J
DJ
Anthony
C
C
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D015153:Blotting, Western; D003432:Cross-Linking Reagents / Q000494:pharmacology; D003574:Cytochrome c Group / Q000302:isolation & purification / Q000378:metabolism*; D006088:Gram-Negative Aerobic Bacteria / Q000201:enzymology*; D007096:Imidoesters / Q000494:pharmacology; D007202:Indicators and Reagents; D007700:Kinetics; D008239:Lysine; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D010658:Phenylglyoxal / Q000031:analogs & derivatives / Q000494:pharmacology; D012689:Sequence Homology, Nucleic Acid; D013050:Spectrometry, Fluorescence; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology
D003432:Cross-Linking Reagents; D003574:Cytochrome c Group; D007096:Imidoesters; D007202:Indicators and Reagents; D046911:Macromolecular Substances; C005905:methyl 4-mercaptobutyrimidate; D014302:Trinitrobenzenesulfonic Acid; C058498:cytochrome C(L); D000429:Alcohol Oxidoreductases; C019641:alcohol dehydrogenase (acceptor); D008239:Lysine; D010658:Phenylglyoxal