title: Monomeric alcohol oxidase is preferentially digested by a novel protease from Candida boidinii.
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.affiliation: Department of Pharmacology, University of Texas Southwestern Medical School, Dallas, TX 75390-9041, USA.
authors.forename: Mary Q
authors.initials: MQ
authors.lastname: Stewart
authors.forename: Ralf
authors.initials: R
authors.lastname: van Dijk
authors.forename: Marten
authors.initials: M
authors.lastname: Veenhuis
authors.forename: Joel M
authors.initials: JM
authors.lastname: Goodman
mesh_terms: D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D002175:Candida / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D003374:Coumarins; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D010455:Peptides / Q000737:chemistry; D020675:Peroxisomes / Q000201:enzymology; D011480:Protease Inhibitors / Q000494:pharmacology; D017510:Protein Folding; D013379:Substrate Specificity
chemical_list: D003374:Coumarins; D007202:Indicators and Reagents; D010455:Peptides; D011480:Protease Inhibitors; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D010450:Endopeptidases; C028743:7-amino-4-methylcoumarin