Identity of soluble thiamine-binding protein with thiamine repressible acid phosphatase in Saccharomyces cerevisiae.
Two secretory glycoproteins of S. cerevisiae, a soluble thiamine-binding protein and a thiamine-repressible acid phosphatase, were shown to be repressed to a similar extent by excess thiamine in the growth medium. Thiamine-repressible acid phosphatase was co-purified throughout the purification of the soluble thiamine-binding protein. Purified and deglycosylated soluble thiamine-binding proteins exhibited both thiamine-binding and acid phosphatase activities on non-denaturing polyacrylamide gel electrophoresis. Heat treatment of the purified soluble thiamine-binding protein caused a decrease in both activities with a similar inactivation profile. Two thiamine-repressible acid phosphatase-defective mutants isolated were found to be also defective in soluble thiamine-binding activity. The uptake of [14C]thiamine phosphate esters, such as thiamine monophosphate and thiamine pyrophsphate, was remarkably impaired in the mutant cells, whereas the uptake of [14C]thiamine by the mutant was almost the same with that by the parent strain. From these results, it was concluded that the soluble thiamine-binding protein is identical to the thiamine-repressible acid phosphatase in S. cerevisiae, which is involved in the hydrolysis of exogenous thiamine phosphate esters in the periplasmic space prior to the uptake of their thiamine moiety by yeast cells.
Nosaka
K
K
Department of Biochemistry, Kyoto Prefectural University of Medicine, Japan.
Nishimura
H
H
Iwashima
A
A
D000135:Acid Phosphatase / Q000032:analysis* / Q000302:isolation & purification; D002352:Carrier Proteins / Q000032:analysis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013831:Thiamine / Q000378:metabolism*
D002352:Carrier Proteins; D029701:Saccharomyces cerevisiae Proteins; C026989:thiamine-binding protein; D000135:Acid Phosphatase; C488913:PHO3 protein, S cerevisiae; C488916:PHO5 protein, S cerevisiae; D013831:Thiamine