Characterization of the NifS-like domain of ABA3 from Arabidopsis thaliana provides insight into the mechanism of molybdenum cofactor sulfuration

J Biol Chem. 2005 Feb 11;280(6):4213-8. doi: 10.1074/jbc.M411195200. Epub 2004 Nov 22.

Abstract

The molybdenum cofactor sulfurase ABA3 from Arabidopsis thaliana specifically regulates the activity of the molybdenum enzymes aldehyde oxidase and xanthine dehydrogenase by converting their molybdenum cofactor from the desulfo-form into the sulfo-form. ABA3 is a two-domain protein with an NH2-terminal domain sharing significant similarities to NifS proteins that catalyze the decomposition of l-cysteine to l-alanine and elemental sulfur for iron-sulfur cluster synthesis. Although different in its physiological function, the mechanism of ABA3 for sulfur mobilization was found to be similar to NifS proteins. The protein binds a pyridoxal phosphate cofactor and a substrate-derived persulfide intermediate, and site-directed mutagenesis of strictly conserved binding sites for the cofactor and the persulfide demonstrated that they are essential for molybdenum cofactor sulfurase activity. In vitro, the NifS-like domain of ABA3 activates aldehyde oxidase and xanthine dehydrogenase in the absence of the C-terminal domain, but in vivo, the C-terminal domain is required for proper activation of both target enzymes. In addition to its cysteine desulfurase activity, ABA3-NifS also exhibits selenocysteine lyase activity. Although l-selenocysteine is unlikely to be a natural substrate for ABA3, it is decomposed more efficiently than l-cysteine. Besides mitochondrial AtNFS1 and plastidial AtNFS2, which are both proposed to be involved in iron-sulfur cluster formation, ABA3 is proposed to be a third and cytosolic NifS-like cysteine desulfurase in A. thaliana. However, the sulfur transferase activity of ABA3 is used for post-translational activation of molybdenum enzymes rather than for iron-sulfur cluster assembly.

MeSH terms

  • Aldehyde Oxidase / metabolism
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins
  • Bacterial Proteins / chemistry
  • Binding Sites
  • Catalysis
  • Coenzymes / chemistry*
  • Cysteine / chemistry
  • Cytosol / chemistry
  • Fluorescent Dyes / pharmacology
  • Genetic Vectors
  • Iron-Sulfur Proteins / chemistry
  • Kinetics
  • Lyases / chemistry
  • Lysine / chemistry
  • Metalloproteins / chemistry*
  • Molybdenum / chemistry*
  • Mutagenesis, Site-Directed
  • Naphthalenesulfonates / pharmacology
  • Pichia / metabolism
  • Plant Proteins / chemistry
  • Protein Binding
  • Protein Structure, Tertiary
  • Pteridines / chemistry*
  • Pyridoxal Phosphate / chemistry
  • Selenocysteine / chemistry
  • Spectrophotometry
  • Substrate Specificity
  • Sulfides / chemistry
  • Sulfurtransferases / chemistry*
  • Sulfurtransferases / metabolism
  • Xanthine Dehydrogenase / metabolism

Substances

  • Arabidopsis Proteins
  • Bacterial Proteins
  • Coenzymes
  • Fluorescent Dyes
  • Iron-Sulfur Proteins
  • Metalloproteins
  • Naphthalenesulfonates
  • Plant Proteins
  • Pteridines
  • Sulfides
  • nifS protein, Bacteria
  • persulfides
  • Selenocysteine
  • Pyridoxal Phosphate
  • 1,5-I-AEDANS
  • Molybdenum
  • molybdenum cofactor
  • Xanthine Dehydrogenase
  • Aldehyde Oxidase
  • ABA3 protein, Arabidopsis
  • Sulfurtransferases
  • Lyases
  • selenocysteine lyase
  • Lysine
  • Cysteine