A lysine conserved in the monoamine oxidase family is involved in oxidation of the reduced flavin in mouse polyamine oxidase

Michelle Henderson Pozzi, Paul F. Fitzpatrick

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Lysine 315 of mouse polyamine amine oxidase corresponds to a lysine residue that is conserved in the flavoprotein amine oxidases of the monoamine oxidase structural family. In several structures, this lysine residue forms a hydrogen bond to a water molecule that is hydrogen-bonded to the flavin N(5). Mutation of Lys315 in polyamine oxidase to methionine was previously shown to have no effect on the kinetics of the reductive half-reaction of the enzyme (M. Henderson Pozzi, V. Gawandi, P.F. Fitzpatrick, Biochemistry 48 (2009) 1508-1516). In contrast, the mutation does affect steps in the oxidative half-reaction. The kcat value is unaffected by the mutation; this kinetic parameter likely reflects product release. At pH 10, the kcat/Km value for oxygen is 25-fold lower in the mutant enzyme. The kcat/KO2 value is pH-dependent for the wild-type enzyme, decreasing below a pKa of 7.0, while this kinetic parameter for the mutant enzyme is pH-independent. This is consistent with the neutral form of Lys315 being required for more rapid flavin oxidation. The solvent isotope effect on the kcat/KO2 value increases from 1.4 in the wild-type enzyme to 1.9 in the mutant protein, and the solvent inventory changes from linear to bowed. The effects of the mutation can be explained by the lysine orienting the bridging water so that it can accept the proton from the flavin N(5) during flavin oxidation. In the mutant enzyme the lysine amine would be replaced by a water chain.

Original languageEnglish (US)
Pages (from-to)83-88
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume498
Issue number2
DOIs
StatePublished - Jun 2010

Keywords

  • Enzyme kinetics
  • Enzyme mechanisms
  • Flavoproteins
  • Isotope effects

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Fingerprint Dive into the research topics of 'A lysine conserved in the monoamine oxidase family is involved in oxidation of the reduced flavin in mouse polyamine oxidase'. Together they form a unique fingerprint.

Cite this