Secretion of an active nonglycosylated form of the repressible acid phosphatase of Saccharomyces cerevisiae in the presence of tunicamycin at low temperatures.
The role of mannan chains in the formation and secretion of active acid phosphatase of yeast (Saccharomyces cerevisiae), a repressible cell surface mannoprotein, was studied in yeast protoplast systems by using tunicamycin at various temperatures. At 30 degrees C, tunicamycin-treated protoplasts did not produce active acid phosphatase; however, at 25 or 20 degrees C they formed and secreted active enzyme. This form of acid phosphatase gave 59-, 57-, and 55-kDa bands on SDS-PAGE which neither bound to concanavalin A Sepharose, nor changed in molecular weight upon treatment with endoglycosidase H, indicating that the peptides are nonglycosylated. The nonglycosylated form, like its glycosylated counterpart, is a dimer on the basis of gel permeation chromatography. The Km for para-nitrophenyl-phosphate and Ki for inorganic phosphate of both glycosylated and nonglycosylated acid phosphatases were almost the same. These results suggested that 1) the conformation of the nonglycosylated acid phosphatase secreted at low temperatures is probably identical with that of the glycosylated one, and 2) the conformation of acid phosphatase is very important for its secretion. The rate of intracellular transport of nonglycosylated acid phosphatase is about one-fourth that of the glycosylated enzyme, indicating that glycosylation facilitates the transport of acid phosphatase proteins.
Mizunaga
T
T
Department of Agricultural Chemistry, Faculty of Agriculture, University of Tokyo.
Izawa
M
M
Ikeda
K
K
Maruyama
Y
Y
D000135:Acid Phosphatase / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006596:Hexosaminidases / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D011523:Protoplasts; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012685:Sepharose / Q000031:analogs & derivatives / Q000378:metabolism; D013696:Temperature; D014415:Tunicamycin / Q000494:pharmacology*
C028326:concanavalin A-sepharose; D014415:Tunicamycin; D012685:Sepharose; D000135:Acid Phosphatase; D006596:Hexosaminidases