title: [DNA-[N4-cytosine]-methyltransferase from Bacillus amyloliquefaciens: mechanism of action derived from steady state kinetics].
abstract: Kinetic analysis of methyl group transfer from S-adenosyl-L-methionine (SAM) to the 5'-GGATCC recognition site catalyzed by the DNA-[N4-cytosine]-methyltransferase from Bacillus amyloliquefaciens [EC 2.1.1.113] has shown that the dependence of the rate of methylation of the 20-meric substrate duplex on SAM and DNA concentration are normally hyperbolic, and the maximal rate is attained upon enzyme saturation with both substrates. No substrate inhibition is observed even at concentrations many times higher than the Km values (0.107 microM for DNA and 1.45 microM for SAM), which means that no nonreactive enzyme-substrate complexes are formed during the reaction. The overall pattern of product inhibition corresponds to an ordered steady-state mechanism following the sequence SAM decreases DNA decreases metDNA increases SAH increases (S-adenosyl-L-homocysteine). However, more detailed numerical analysis of the aggregate experimental data admits an alternative order of substrate binding, DNA decreases SAM decreases, though this route is an order of magnitude slower.
authors.affiliation: Institute of Molecular Biology, State Research Center for Virology and Biotechnology VECTOR, Kol'tsovo, Novosibirsk Oblast, 630559 Russia.
authors.forename: V V
authors.initials: VV
authors.lastname: Zinov'ev
authors.forename: A A
authors.initials: AA
authors.lastname: Evdokimov
authors.forename: E G
authors.initials: EG
authors.lastname: Malygin
mesh_terms: D001407:Bacillus / Q000201:enzymology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D015257:DNA-Cytosine Methylases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D008956:Models, Chemical; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism
chemical_list: D004269:DNA, Bacterial; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; D015257:DNA-Cytosine Methylases