Yeast mannosyl transferases requiring dolichyl phosphate and dolichyl phosphate mannose as substrate. Partial purification and characterization of the solubilized enzyme

Eur J Biochem. 1980 Apr;105(3):509-15. doi: 10.1111/j.1432-1033.1980.tb04526.x.


The first mannosyl unit of manno-oligosaccharides of fungal mannoproteins is transferred in a dolichyl-phosphate-dependent reaction sequence to serine/threonine residues of the protein. The two membrane-bound enzymes catalyzing this transfer in the yeast Saccharomyces cerevisiae have been solubilized by detergents. The enzyme transferring mannose from guanosine diphosphate mannose to dolichyl phosphate has been purified 18-fold when based on membrane protein and 140-fold when based on total cell protein. The enzyme transferring mannose from dolichyl phosphate mannose to protein has been purified 48-fold and 380-fold, respectively. A HCl-treated cell-wall mannoprotein from yeast served as acceptor protein for the second enzyme. The solubilized enzyme catalyzing the formation of dolichyl diphosphate mannose has a Km for guanosine diphosphate mannose of 7 x 10(-6) M and is saturated with about 0.15 mM yeast dolichyl phosphate. The metal requirement, pH-optima, and the detergent concentration necessary for optimal activity have been determined for both solubilized enzymes.

MeSH terms

  • Chromatography
  • Dolichol Monophosphate Mannose / metabolism*
  • Dolichol Phosphates / metabolism*
  • Hexosyltransferases / metabolism*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Magnesium / metabolism
  • Manganese / metabolism
  • Mannosyltransferases / isolation & purification
  • Mannosyltransferases / metabolism*
  • Octoxynol
  • Polyethylene Glycols
  • Polyisoprenyl Phosphate Sugars / metabolism*
  • Polyisoprenyl Phosphates / metabolism*
  • Saccharomyces cerevisiae / enzymology*
  • Solubility


  • Dolichol Phosphates
  • Polyisoprenyl Phosphate Sugars
  • Polyisoprenyl Phosphates
  • Polyethylene Glycols
  • Manganese
  • Dolichol Monophosphate Mannose
  • Octoxynol
  • Hexosyltransferases
  • Mannosyltransferases
  • Magnesium