Saccharomyces cerevisiae structural cell wall mannoprotein.
A novel mannoprotein fraction with an average molecular weight of 180 000 has been isolated from Saccharomyces cerevisiae mnn9 mutant cell wall that was solubilized by beta-glucanase digestion. The same material could be extracted from purified wall fragments with 1% sodium dodecyl sulfate. The protein component, 12% by weight, is rich in proline, whereas the carbohydrate, mainly mannose, is about evenly distributed between asparagine and hydroxyamino acids. Endoglucosaminidase H digestion of the isolated mannoprotein reduced its average molecular weight to 150 000, but the mannoprotein, while still embedded in the cell wall, was inaccessible to the enzyme. Biosynthesis and translocation of the mannoprotein were investigated by following incorporation of [3H]proline into this fraction. In the presence of tunicamycin, both mnn9 and wild-type X2180 cells made a mannoprotein fraction with an average molecular weight of 140 000, whereas in the absence of the glycosylation inhibitor, the mnn9 mutant made material with a molecular weight of 180 000 and the mannoprotein made by wild-type cells was too large to penetrate the polyacrylamide gel. Although the cell wall mannoprotein was resistant to heat and proteolytic enzymes, attempts to isolate the carbohydrate-free component failed to yield any characteristic peptide material.
D000596:Amino Acids / Q000032:analysis; D002473:Cell Wall / Q000032:analysis; D006023:Glycoproteins / Q000302:isolation & purification*; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000302:isolation & purification*; D008970:Molecular Weight; D009154:Mutation; D009844:Oligosaccharides / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000235:genetics
D000596:Amino Acids; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009844:Oligosaccharides; C025363:mannoproteins