| Title: | Characterization of novel peptide agonists of the alpha mating factor of Saccharomyces cerevisiae |
| Author(s): | Siegel EG; Gunther R; Schafer H; Folsch UR; Schmidt WE; |
| Address: | "Laboratory of Molecular Gastroenterology, Gastrointestinal Unit, 1st Department of Medicine, Christian-Albrechts-University of Kiel, Kiel, 24105, Germany" |
| ISSN/ISBN: | 0003-2697 (Print) 0003-2697 (Linking) |
| Abstract: | "Alpha-factor [WHWLQLKPGQPMY], a secreted tridecapeptide pheromone, is required for mating between the a- and alpha-haploid mating types of Saccharomyces cerevisiae (MATa, MATalpha). New analogues of alpha-factor were synthesized and evaluated by morphogenesis assays and receptor binding studies. The Y(0)Nle(12)F(13) analogue [YWHWLQLKPGQPNleF] (MFN5) caused growth arrest and morphological alteration in MATa cells in a fashion identical to that of the native pheromone. Binding of (125)I-labeled MFN5 was saturable, and reversible as shown by equipotent label displacement by MFN5 and native alpha-mating factor. Scatchard analysis of equilibrium binding data on plasma membranes and intact cells indicated the existence of a single high-affinity binding site (K(d) = 6.4 x 10(-8)). Specific binding of (125)I-labeled MFN5 was significantly reduced by guanosine nucleotides. Affinity cross-linking of (125)I-labeled MFN5 to MATa cell membranes identified a specifically labeled 49-kDa protein. The novel synthetic alpha-factor analogue MFN5 can be easily iodinated and used as a probe for the alpha-factor receptor" |
| Keywords: | Affinity Labels Amino Acid Sequence Binding Sites Biological Assay/methods Cell Membrane/metabolism Cross-Linking Reagents Iodine Radioisotopes Mating Factor Molecular Sequence Data Peptides/*agonists/*analysis/chemistry/metabolism/pharmacology Saccharomy; |
| Notes: | "MedlineSiegel, E G Gunther, R Schafer, H Folsch, U R Schmidt, W E eng 1999/11/05 Anal Biochem. 1999 Nov 1; 275(1):109-15. doi: 10.1006/abio.1999.4289" |
