Biosynthesis of yeast mannan. Properties of a mannosylphosphate transferase in Saccharomyces cerevisiae.
A homogenate of mechanically broken, freshly grown Saccharomyces cerevisiae X2180 cells catalyzes the transfer of mannosylphosphate units from guanosine diphosphate mannose to reduced alpha1 leads to 2-[3H]mannotetraose to yield reduced mannosylphosphoryl [3H]-mannotetraose. The product is analogous in structure to the phosphorylated mannan side chains, which suggests that the enzymic activity is involved in mannoprotein biosynthesis in the intact cell. The mannosylphosphate transferase activity, localized in a membrane fraction obtained by differential centrifugation at 100,000 x g, was solubilized by Triton X-155 and purified 250-fold by ammonium sulfate precipitation and by ion exchange and gell filtration chromatographies. The enzyme requires MN2+ OR Co2+ ions for activity and is stimulated by various detergents. The mnn2 and mnn3 mannan mutants of S. cerevisiae possess normal levels of mannosylphosphate transferase activity, whereas the mnn4 mutant cells contain very low, if any, activity. This is consistent with a previous conclusion that the mnn4 mutation affects the mannosylphosphate transferase activity, whereas the mnn2 and mnn3 strains possess phosphate-deficient mannans because they are unable to synthesize the appropriate side chain precursors. A new mannan mutant class with the mnn4 chemotype was isolated, but the mutation proved to be recessive and nonallelic with the mnn4 locus. This new locus is designated mnn6.
D004287:Dolichol Monophosphate Mannose / Q000378:metabolism; D006602:Hexosyltransferases / Q000378:metabolism*; D008351:Mannans / Q000096:biosynthesis*; D008360:Mannosephosphates / Q000378:metabolism; D008364:Mannosyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009154:Mutation; D011134:Polysaccharides / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D008351:Mannans; D008360:Mannosephosphates; D011134:Polysaccharides; D004287:Dolichol Monophosphate Mannose; D006602:Hexosyltransferases; D008364:Mannosyltransferases