This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Manfredi, J. P. Articles by Broach, J. R. Search for Related Content PubMed PubMed Citation Articles by Manfredi, J. P. Articles by Broach, J. R.

 Previous Article  |  Next Article 

Mol. Cell. Biol., Sep 1996, 4700-4709, Vol 16, No. 9
Copyright © 1996, American Society for Microbiology

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

JP Manfredi, C Klein, JJ Herrero, DR Byrd, J Trueheart, WT Wiesler, DM Fowlkes and JR Broach
Cadus Pharmaceutical Corporation, Tarrytown, New York 10591-6705, USA.

alpha-Factor, a 13-amino-acid pheromone secreted by haploid alpha cells of Saccharomyces cerevisiae, binds to Ste2p, a seven-transmembrane, G- protein-coupled receptor present on haploid alpha cells, to activate a signal transduction pathway required for conjugation and mating. To determine the structural requirements for alpha-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-protein-coupled receptors heterologously expressed in S. cerevisiae.

This article has been cited by other articles:

• Ishii, J., Tanaka, T., Matsumura, S., Tatematsu, K., Kuroda, S., Ogino, C., Fukuda, H., Kondo, A. (2008). Yeast-Based Fluorescence Reporter Assay of G Protein-coupled Receptor Signalling for Flow Cytometric Screening: FAR1-Disruption Recovers Loss of Episomal Plasmid Caused by Signalling in Yeast. J Biochem 143: 667-674 [Abstract] [Full Text]  
• Ault, A. D., Broach, J. R. (2006). Creation of GPCR-based chemical sensors by directed evolution in yeast. Protein Eng Des Sel 19: 1-8 [Abstract] [Full Text]  
• Sachpatzidis, A., Benton, B. K., Manfredi, J. P., Wang, H., Hamilton, A., Dohlman, H. G., Lolis, E. (2003). Identification of Allosteric Peptide Agonists of CXCR4. J. Biol. Chem. 278: 896-907 [Abstract] [Full Text]  
• Miret, J. J., Rakhilina, L., Silverman, L., Oehlen, B. (2002). Functional Expression of Heteromeric Calcitonin Gene-related Peptide and Adrenomedullin Receptors in Yeast. J. Biol. Chem. 277: 6881-6887 [Abstract] [Full Text]  
• Chambers, J. K., Macdonald, L. E., Sarau, H. M., Ames, R. S., Freeman, K., Foley, J. J., Zhu, Y., McLaughlin, M. M., Murdock, P., McMillan, L., Trill, J., Swift, A., Aiyar, N., Taylor, P., Vawter, L., Naheed, S., Szekeres, P., Hervieu, G., Scott, C., Watson, J. M., Murphy, A. J., Duzic, E., Klein, C., Bergsma, D. J., Wilson, S., Livi, G. P. (2000). A G Protein-coupled Receptor for UDP-glucose. J. Biol. Chem. 275: 10767-10771 [Abstract] [Full Text]  
• Tanaka, T., Nakayama, K., Machida, K., Taniguchi, M. (2000). Long-chain alkyl ester of AMP acts as an antagonist of glucose-induced signal transduction that mediates activation of plasma membrane proton pump in Saccharomyces cerevisiae. Microbiology 146: 377-384 [Abstract] [Full Text]  
• Parandoosh, Z. (1997). Cell-Based Assays. J Biomol Screen 2: 201-202