Prm1 functions as a disulfide-linked complex in yeast mating

Valerie N. Olmo, Eric Grote

Research output: Contribution to journalArticle

Abstract

Prm1 is a pheromone-induced membrane glycoprotein that promotes plasma membrane fusion in yeast mating pairs. HA-Prm1 migrates at twice its expected molecular weight on nonreducing SDS-PAGE gels and coprecipitates with Prm1-TAP, indicating that Prm1 is a disulfide-linked homodimer. The N terminus of a plasma membrane-localized GFP-Prm1 endocytic mutant projects into the cytoplasm, where it is protected from low pH quenching in live cells and from external protease in spheroplasts. In a revised topological map, Prm1 has four transmembrane domains and two large extracellular loops. Mutation of all four cysteines in the extracellular loops blocked disulfide bond formation and destabilized the Prm1 homodimer without preventing Prm1 transport to contact sites in mating pairs. Cys120 in loop 1 and Cys545 in loop 2 form disulfide cross-links in the Prm1 homodimer and are required for fusion activity. Cys 120 lies between a hydrophobic segment formerly thought to be a transmembrane domain and an amphipathic helix. An interaction between either of these regions and the opposing membrane could promote fusion.

Original languageEnglish (US)
Pages (from-to)2274-2283
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number4
DOIs
StatePublished - Jan 22 2010

Fingerprint

Disulfides
Yeast
Fusion reactions
Yeasts
Cell membranes
Cell Membrane
Spheroplasts
Membrane Fusion
Pheromones
Membrane Glycoproteins
Cysteine
Polyacrylamide Gel Electrophoresis
Quenching
Cytoplasm
Peptide Hydrolases
Molecular Weight
Gels
Molecular weight
Membranes
Mutation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Prm1 functions as a disulfide-linked complex in yeast mating. / Olmo, Valerie N.; Grote, Eric.

In: Journal of Biological Chemistry, Vol. 285, No. 4, 22.01.2010, p. 2274-2283.

Research output: Contribution to journalArticle

Olmo, Valerie N. ; Grote, Eric. / Prm1 functions as a disulfide-linked complex in yeast mating. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 4. pp. 2274-2283.
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