title: Studies on porphobilinogen-deaminase from Saccharomyces cerevisiae.
abstract: Porphobilinogen-deaminase from Saccharomyces cerevisiae has been isolated and partially purified 80- and 230-fold in the absence or presence of phenylmethylsulphonyl fluoride, respectively. Some properties of the isolated enzyme were studied. Porphyrin formation was linear with time and protein concentration. Optimum pH was about 7.5-7.8. Molecular mass of the protein was 30,000 +/- 3000 Dalton when the enzyme was purified in the presence of phenylmethylsulphonyl fluoride. A less active and unstable 20,000 Da molecular mass species was obtained when purification was performed in the absence of the protease inhibitor. Porphobilinogen-deaminase exhibited classical Michaelis-Menten kinetics. The apparent Km for uroporphyrinogen formation was 19 microM; Vmax was 3.6 nmol uroporphyrin/h and the Hill coefficient was n = 1. Also the action of several reagents on the activity was studied. Protective thiol agents had no effect. Heavy metals inhibited both porphyrin formation and porphobilinogen consumption, but known sulphydryl inactivating chemicals inhibit the former without modifying the latter. Ammonium ions had no effect on the activity while hydroxylamine completely inhibited both porphyrin formation and porphobilinogen consumption.
authors.affiliation: Centro de Investigaciones sobre Porfirinas y Porfirias-CIPYP (CONICET, F.C.E. y N., UBA)-Ciudad Universitaria, Buenos Aires, Argentine.
authors.forename: S R
authors.initials: SR
authors.lastname: Correa García
authors.forename: M V
authors.initials: MV
authors.lastname: Rossetti
authors.forename: A M
authors.initials: AM
authors.lastname: Batlle
mesh_terms: D002850:Chromatography, Gel / Q000379:methods; D006863:Hydrogen-Ion Concentration; D011163:Hydroxymethylbilane Synthase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D010664:Phenylmethylsulfonyl Fluoride; D011163:Hydroxymethylbilane Synthase