This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Manfredi, J. P.
Right arrow Articles by Broach, J. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Manfredi, J. P.
Right arrow 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  
  • Gillece, P., Pilon, M., Romisch, K. (2000). The protein translocation channel mediates glycopeptide export across the endoplasmic reticulum membrane. Proc. Natl. Acad. Sci. USA 97: 4609-4614 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»