Yeast α mating factor structure-activity relationship derived from genetically selected peptide agonists and antagonists of Ste2p

John P. Manfredi, Christine Klein, Juan J. Herrero, Devon R. Byrd, Joshua Trueheart, William T. Wiesler, Dana M. Fowlkes, James Broach

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

α-Factor, a 13-amino-acid pheromone secreted by haploid α cells of Saccharomyces cerevisiae, binds to Ste2p, a seven-transmembrane, G-protein- coupled receptor present on haploid a cells, to activate a signal transduction pathway required for conjugation and mating. To determine the structural requirements for α-factor activity, we developed a genetic screen to identify from random and semirandom libraries novel peptides that function as agonists or antagonists of Ste2p. The selection scheme was based on autocrine strains constructed to secrete random peptides and respond by growth to those that were either agonists or antagonists of Ste2p. Analysis of a number of peptides obtained by this selection procedure indicates that Trp1, Trp3, Pro8, and Gly9 are important for agonist activity specifically. His2, Leu4, Leu6, Pro10, a hydrophobic residue 12, and an aromatic residue 13 are important for both agonist and antagonist activity. Our results also show that activation of Ste2p can be achieved with novel, unanticipated combinations of amino acids. Finally, the results suggest the utility of this selection scheme for identifying novel ligands for mammalian G-prntein- coupled receptors heterologously expressed in S. cerevisiae.

Original languageEnglish (US)
Pages (from-to)4700-4709
Number of pages10
JournalMolecular and cellular biology
Volume16
Issue number9
StatePublished - Aug 27 1996

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Mating Factor
Haploidy
Structure-Activity Relationship
Saccharomyces cerevisiae
Yeasts
Amino Acids
Peptide Library
Peptides
Pheromones
Signal Transduction
Ligands
Growth

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Manfredi, John P. ; Klein, Christine ; Herrero, Juan J. ; Byrd, Devon R. ; Trueheart, Joshua ; Wiesler, William T. ; Fowlkes, Dana M. ; Broach, James. / Yeast α mating factor structure-activity relationship derived from genetically selected peptide agonists and antagonists of Ste2p. In: Molecular and cellular biology. 1996 ; Vol. 16, No. 9. pp. 4700-4709.
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abstract = "α-Factor, a 13-amino-acid pheromone secreted by haploid α cells of Saccharomyces cerevisiae, binds to Ste2p, a seven-transmembrane, G-protein- coupled receptor present on haploid a cells, to activate a signal transduction pathway required for conjugation and mating. To determine the structural requirements for α-factor activity, we developed a genetic screen to identify from random and semirandom libraries novel peptides that function as agonists or antagonists of Ste2p. The selection scheme was based on autocrine strains constructed to secrete random peptides and respond by growth to those that were either agonists or antagonists of Ste2p. Analysis of a number of peptides obtained by this selection procedure indicates that Trp1, Trp3, Pro8, and Gly9 are important for agonist activity specifically. His2, Leu4, Leu6, Pro10, a hydrophobic residue 12, and an aromatic residue 13 are important for both agonist and antagonist activity. Our results also show that activation of Ste2p can be achieved with novel, unanticipated combinations of amino acids. Finally, the results suggest the utility of this selection scheme for identifying novel ligands for mammalian G-prntein- coupled receptors heterologously expressed in S. cerevisiae.",
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Manfredi, JP, Klein, C, Herrero, JJ, Byrd, DR, Trueheart, J, Wiesler, WT, Fowlkes, DM & Broach, J 1996, 'Yeast α mating factor structure-activity relationship derived from genetically selected peptide agonists and antagonists of Ste2p', Molecular and cellular biology, vol. 16, no. 9, pp. 4700-4709.

Yeast α mating factor structure-activity relationship derived from genetically selected peptide agonists and antagonists of Ste2p. / Manfredi, John P.; Klein, Christine; Herrero, Juan J.; Byrd, Devon R.; Trueheart, Joshua; Wiesler, William T.; Fowlkes, Dana M.; Broach, James.

In: Molecular and cellular biology, Vol. 16, No. 9, 27.08.1996, p. 4700-4709.

Research output: Contribution to journalArticle

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T1 - Yeast α mating factor structure-activity relationship derived from genetically selected peptide agonists and antagonists of Ste2p

AU - Manfredi, John P.

AU - Klein, Christine

AU - Herrero, Juan J.

AU - Byrd, Devon R.

AU - Trueheart, Joshua

AU - Wiesler, William T.

AU - Fowlkes, Dana M.

AU - Broach, James

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AB - α-Factor, a 13-amino-acid pheromone secreted by haploid α cells of Saccharomyces cerevisiae, binds to Ste2p, a seven-transmembrane, G-protein- coupled receptor present on haploid a cells, to activate a signal transduction pathway required for conjugation and mating. To determine the structural requirements for α-factor activity, we developed a genetic screen to identify from random and semirandom libraries novel peptides that function as agonists or antagonists of Ste2p. The selection scheme was based on autocrine strains constructed to secrete random peptides and respond by growth to those that were either agonists or antagonists of Ste2p. Analysis of a number of peptides obtained by this selection procedure indicates that Trp1, Trp3, Pro8, and Gly9 are important for agonist activity specifically. His2, Leu4, Leu6, Pro10, a hydrophobic residue 12, and an aromatic residue 13 are important for both agonist and antagonist activity. Our results also show that activation of Ste2p can be achieved with novel, unanticipated combinations of amino acids. Finally, the results suggest the utility of this selection scheme for identifying novel ligands for mammalian G-prntein- coupled receptors heterologously expressed in S. cerevisiae.

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