title: Kinetic and spectral parameters of interaction of Citrobacter freundii methionine γ-lyase with amino acids.
abstract: Kinetic parameters of Citrobacter freundii methionine γ-lyase were determined with substrates in γ-elimination reactions as well as the inhibition of the enzyme in the γ-elimination of L-methionine by amino acids with different structure. The data indicate an important contribution of the sulfur atom and methylene groups to the efficiency of binding of substrates and inhibitors. The rate constants of the enzyme-catalyzed exchange of C-α- and C-β-protons with deuterium were determined, as well as the kinetic isotope effect of the deuterium label in the C-α-position of inhibitors on the rate of exchange of their β-protons. Neither stereoselectivity in the β-proton exchange nor noticeable α-isotope effect on the exchange rates of β-protons was found. The ionic and tautomeric composition of the external Schiff base of methionine γ-lyase was determined. Spectral characteristics (absorption and circular dichroism spectra) of complexes with substrates and inhibitors were determined. The spectral and kinetic data indicate that deamination of aminocrotonate should be the rate-determining stage of the enzymatic reaction.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.
authors.forename: E A
authors.initials: EA
authors.lastname: Morozova
authors.forename: N P
authors.initials: NP
authors.lastname: Bazhulina
authors.forename: N V
authors.initials: NV
authors.lastname: Anufrieva
authors.forename: D V
authors.initials: DV
authors.lastname: Mamaeva
authors.forename: Y V
authors.initials: YV
authors.lastname: Tkachev
authors.forename: S A
authors.initials: SA
authors.lastname: Streltsov
authors.forename: V P
authors.initials: VP
authors.lastname: Timofeev
authors.forename: N G
authors.initials: NG
authors.lastname: Faleev
authors.forename: T V
authors.initials: TV
authors.lastname: Demidkina
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000737:chemistry* / Q000378:metabolism; D016971:Citrobacter freundii / Q000201:enzymology*; D007700:Kinetics; D013379:Substrate Specificity / Q000502:physiology
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D013437:Carbon-Sulfur Lyases; C021558:L-methionine gamma-lyase