title: Purification and characterization of methionine gamma-lyase from Trichomonas vaginalis.
abstract: Methionine gamma-lyase (EC 4.4.1.11) was purified to homogeneity from the anaerobic protozoan parasite Trichomonas vaginalis by a series of f.p.l.c. procedures. The enzyme catalyses alpha gamma- and alpha beta-elimination reactions of a number of derivatives of methionine and cysteine. It also catalyses gamma-replacement reactions of the thiomethyl group of methionine, homocysteine and ethionine to yield the corresponding S-substituted homocysteine derivative. The enzyme is pyridoxal 5'-phosphate-dependent, has a native molecular mass of approx. 160 kDa and consists of four apparently identical subunits of molecular mass 43-45 kDa. The absorption spectrum of the enzyme is typical of those obtained for other pyridoxal 5'-phosphate-dependent enzymes, and the holoenzyme can be resolved to the apoenzyme by incubation with hydroxylamine and reconstituted by addition of the cofactor. The enzyme activity is significantly affected by carbonyl and thiol reagents, is competitively inhibited by a number of substrate analogues and is completely inactivated by the suicide inhibitor DL-propargylglycine. The T. vaginalis enzyme is similar, in terms of activity and properties, to the enzymes found in a number of species of bacteria that metabolize methionine under anaerobic conditions. It is suggested that methionine catabolism may be of particular importance to the survival of T. vaginalis under microaerophilic conditions in its host.
authors.affiliation: Department of Zoology, University of Glasgow, Scotland, U.K.
authors.forename: B C
authors.initials: BC
authors.lastname: Lockwood
authors.forename: G H
authors.initials: GH
authors.lastname: Coombs
mesh_terms: D000818:Animals; D013437:Carbon-Sulfur Lyases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003523:Cycloserine / Q000494:pharmacology; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D010396:Penicillamine / Q000494:pharmacology; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D014246:Trichomonas vaginalis / Q000201:enzymology*
chemical_list: D006898:Hydroxylamines; D015800:Protozoan Proteins; D019811:Hydroxylamine; D003523:Cycloserine; D013437:Carbon-Sulfur Lyases; C021558:L-methionine gamma-lyase; D010396:Penicillamine