Distinct sequence determinants direct intracellular sorting and modification of a yeast vacuolar protease

Cell. 1987 Mar 13;48(5):875-85. doi: 10.1016/0092-8674(87)90084-5.

Abstract

We have mapped a sequence determinant in the vacuolar glycoprotein carboxypeptidase Y (CPY) that directs intracellular sorting of this enzyme. Through the study of hybrid proteins, consisting of amino-terminal segments of CPY fused to the secretory enzyme invertase, we have found that the N-terminal 50 amino acids of CPY are sufficient to direct delivery of a CPY-Inv hybrid protein to the yeast vacuole. Our data suggest that this 50 amino acid segment of CPY contains two distinct functional domains; an N-terminal signal peptide followed by a segment of 30 amino acids that contains the vacuolar sorting signal. Deletion of this putative vacuole sorting signal from an otherwise wild-type CPY protein leads to missorting of CPY. Furthermore, examination of the Asn-linked oligosaccharides present on CPY and CPY-Inv hybrid proteins suggests that an additional determinant in CPY specifies the extent to which these proteins are glycosylated in the Golgi complex.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Carboxypeptidases / genetics*
  • Carboxypeptidases / metabolism
  • Cathepsin A
  • DNA Restriction Enzymes
  • Glycoside Hydrolases / genetics
  • Glycoside Hydrolases / metabolism
  • Organoids / enzymology*
  • Plasmids
  • Protein Multimerization
  • Protein Processing, Post-Translational*
  • Protein Sorting Signals / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins
  • Vacuoles / enzymology*
  • beta-Fructofuranosidase

Substances

  • Protein Sorting Signals
  • Saccharomyces cerevisiae Proteins
  • DNA Restriction Enzymes
  • Glycoside Hydrolases
  • beta-Fructofuranosidase
  • Carboxypeptidases
  • Cathepsin A
  • PRC1 protein, S cerevisiae
  • serine carboxypeptidase