Differential extraction of N-acetylglucosaminidase and trehalase from the cell envelope of Candida albicans.
Dithiothreitol (DTT) extraction of N-acetylglucosaminidase and trehalase from intact Candida albicans ATCC 10261 cells was monitored as an index of cell envelope porosity during N-acetylglucosamine-induced morphogenesis. Trehalase, which is secreted into the cell envelope during starvation and bud-formation, displayed similar extraction kinetics in starved, germ tube-forming, and bud-forming cells, indicating that the mother cell wall remains largely unchanged during morphogenic outgrowth and that the porosity of bud and mother cell walls is similar. N-acetylglucosaminidase, which is secreted specifically during morphogenesis, was released eightfold more rapidly from germ tube-forming than bud-forming cells, reflecting major differences in porosity between bud and germ tube. In addition, by assaying DTT extracts and extracted cell residues, it was found that the total extracellular N-acetylglucosaminidase activity increased 2- to 2.5-fold during DTT treatment. Thus, DTT unmasks a cryptic form of N-acetylglucosaminidase. The cryptic activity was associated with the cell wall fraction.
Molloy
C
C
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Shepherd
M G
MG
Sullivan
P A
PA
D000118:Acetylglucosaminidase / Q000302:isolation & purification* / Q000378:metabolism; D002176:Candida albicans / Q000201:enzymology* / Q000254:growth & development; D002473:Cell Wall / Q000201:enzymology; D004229:Dithiothreitol; D007700:Kinetics; D014198:Trehalase / Q000302:isolation & purification* / Q000378:metabolism
D014198:Trehalase; D000118:Acetylglucosaminidase; D004229:Dithiothreitol