Veratrol-O-demethylase of Acetobacterium dehalogenans: ATP-dependent reduction of the corrinoid protein.
The anaerobic veratrol O-demethylase mediates the transfer of the methyl group of the phenyl methyl ether veratrol to tetrahydrofolate. The primary methyl group acceptor is the cobalt of a corrinoid protein, which has to be in the +1 oxidation state to bind the methyl group. Due to the negative redox potential of the cob(II)/cob(I)alamin couple, autoxidation of the cobalt may accidentally occur. In this study, the reduction of the corrinoid to the superreduced [Co(I)] state was investigated. The ATP-dependent reduction of the corrinoid protein of the veratrol O-demethylase was shown to be dependent on titanium(III) citrate as electron donor and on an activating enzyme. In the presence of ATP, activating enzyme, and Ti(III), the redox potential versus the standard hydrogen electrode (E (SHE)) of the cob(II)alamin/cob(I)alamin couple in the corrinoid protein was determined to be -290 mV (pH 7.5), whereas E (SHE) at pH 7.5 was lower than -450 mV in the absence of either activating enzyme or ATP. ADP, AMP, or GTP could not replace ATP in the activation reaction. The ATP analogue adenosine-5'-(beta,gamma-imido)triphosphate (AMP-PNP, 2-4 mM) completely inhibited the corrinoid reduction in the presence of ATP (2 mM).
Siebert
Anke
A
Institut für Mikrobiologie, FSU Jena, Lehrstuhl für Angewandte und Okologische Mikrobiologie, Philosophenweg 12, 07743 Jena, Germany.
Schubert
Torsten
T
Engelmann
Tina
T
Studenik
Sandra
S
Diekert
Gabriele
G
D045855:Acetobacterium / Q000201:enzymology*; D000255:Adenosine Triphosphate; D000840:Anisoles / Q000378:metabolism*; D019343:Citric Acid; D003035:Cobalt; D008780:Methyltransferases / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D014805:Vitamin B 12 / Q000031:analogs & derivatives / Q000378:metabolism
D000840:Anisoles; C419387:titanium citrate; C097751:cob(II)alamin; D019343:Citric Acid; D003035:Cobalt; C005299:veratrole; D000255:Adenosine Triphosphate; C060998:anisole; D008780:Methyltransferases; D014805:Vitamin B 12