In vivo processing of nonanchored Yapsin 1 (Yap3p)

Arch Biochem Biophys. 2000 Mar 15;375(2):315-21. doi: 10.1006/abbi.1999.1665.

Abstract

A C-terminally truncated form of yapsin 1 (yeast aspartic protease 3) was overexpressed in yeast and its processing through the secretory pathway was followed by pulse-labeling and immunoprecipitation studies. In the soluble cell extract, three forms of yapsin 1-87, 74, and 18 kDa-were found. Identification of these forms of yapsin 1 using different antisera suggests that the 87-kDa form is pro-yapsin 1, which is processed into two subunits, alpha (18 kDa) and beta (74 kDa), by cleavage at a loop region not found in traditional aspartic proteases. By use of a temperature-sensitive mutant strain, sec18, the generation of the two subunits was found to occur in the endoplasmic reticulum. An active site-mutated yapsin 1 was not processed into the two subunits, suggesting that this process occurs in an autocatalytic manner.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases*
  • Amino Acid Sequence
  • Aspartic Acid Endopeptidases / chemistry
  • Aspartic Acid Endopeptidases / genetics
  • Aspartic Acid Endopeptidases / metabolism*
  • Binding Sites
  • Catalysis
  • Culture Media, Conditioned / chemistry
  • Endoplasmic Reticulum / enzymology
  • Endoplasmic Reticulum / metabolism
  • Fungal Proteins / chemistry
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism
  • Fungal Proteins / physiology
  • Genes, Fungal / genetics
  • Genes, Fungal / physiology
  • Glycosylation
  • Models, Biological
  • Molecular Sequence Data
  • Molecular Weight
  • Mutation / genetics
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism*
  • Precipitin Tests
  • Protein Precursors / chemistry
  • Protein Precursors / genetics
  • Protein Precursors / metabolism*
  • Protein Processing, Post-Translational*
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / enzymology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins*
  • Temperature
  • Vesicular Transport Proteins*

Substances

  • Culture Media, Conditioned
  • Fungal Proteins
  • Peptide Fragments
  • Protein Precursors
  • Saccharomyces cerevisiae Proteins
  • Vesicular Transport Proteins
  • aspartic proteinase A
  • Aspartic Acid Endopeptidases
  • YPS1 protein, S cerevisiae
  • Adenosine Triphosphatases
  • SEC18 protein, S cerevisiae