Monomeric alcohol oxidase is preferentially digested by a novel protease from Candida boidinii.

Academic Article

Abstract

  • A protease activity has been partially purified from peroxisomal matrix fractions of the methylotrophic yeast Candida boidinii. The enzyme migrates as a single peak on a sucrose velocity gradient with an apparent native molecular mass of approximately 80-90 kDa. Activity can be recovered from nonreducing sodium dodecyl sulfate gels as a approximately 20 kDa species, suggesting it is an oligomer. The protein exhibits chymotrypsin-like activity and cleaves the model compound suc-L-L-V-Y-AMC. Additionally, monomers of alcohol oxidase (AO), an abundant protein of C. boidinii peroxisomes, generated in vitro or in pulse-radiolabeled cells, are preferentially sensitive to degradation by the protease. Sensitivity is lost over time in vivo as AO folds and matures into octamers, suggesting that the protease may be involved in these processes.
  • Authors

  • Stewart, Mary
  • van Dijk, Ralf
  • Veenhuis, Marten
  • Goodman, Joel M
  • Status

    Publication Date

  • January 30, 2002
  • Published In

    Keywords

  • Alcohol Oxidoreductases
  • Binding Sites
  • Candida
  • Centrifugation, Density Gradient
  • Chromatography
  • Coumarins
  • Endopeptidases
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Indicators and Reagents
  • Peptides
  • Peroxisomes
  • Protease Inhibitors
  • Protein Folding
  • Substrate Specificity
  • Digital Object Identifier (doi)

    Pubmed Id

  • 11853889
  • Start Page

  • 160
  • End Page

  • 172
  • Volume

  • 1542
  • Issue

  • 1-3