Yeast α Mating Factor Structure-Activity Relationship Derived from Genetically Selected Peptide Agonists and Antagonists of Ste2p
| Autor: | William T. Wiesler, Joshua Trueheart, James R. Broach, Dana M. Fowlkes, Devon R. Byrd, Juan J. Herrero, Christine Klein, John Manfredi |
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| Rok vydání: | 1996 |
| Předmět: |
Microbiological Techniques
Agonist Receptors Peptide medicine.drug_class Molecular Sequence Data Saccharomyces cerevisiae Peptide Biology Structure-Activity Relationship medicine Amino Acid Sequence Amino Acids Selection Genetic Receptor Molecular Biology Peptide sequence chemistry.chemical_classification Base Sequence Cell Biology biology.organism_classification Protein Structure Tertiary Amino acid chemistry Biochemistry Receptors Mating Factor Signal transduction Mating Factor Peptides Function (biology) Research Article Signal Transduction Transcription Factors |
| Zdroj: | Molecular and Cellular Biology. 16:4700-4709 |
| ISSN: | 1098-5549 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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