| Title: | Probing the functional conformation of the tridecapeptide mating pheromone of Saccharomyces cerevisiae through study of disulfide-constrained analogs |
| Author(s): | Xue CB; McKinney A; Lu HF; Jiang Y; Becker JM; Naider F; |
| Address: | "Department of Chemistry, College of Staten Island, City University of New York, USA" |
| DOI: | 10.1111/j.1399-3011.1996.tb01336.x |
| ISSN/ISBN: | 0367-8377 (Print) 0367-8377 (Linking) |
| Abstract: | "Analogs of the Saccharomyces cerevisiae alpha-mating factor, Trp-His-Trp-Leu-Gln-Leu-Lys-Pro-Gly-Gln-Pro-Met-Tyr, where Lys7 and Gln10 were replaced with Cys, Cys(CH3), or Ser, were synthesized using solid-phase procedures on a phenylacetamidomethyl resin. Cyclo7,10[Cys7,X9,Cys10,Nle12]alpha-factor , where X=D-Val, D-Ala, L-Ala and Gly, were prepared by on-resin cyclization using thallic trifluoroacetate in yields of 20-30%. Linear sulfhydryl-containing peptides were generated from their corresponding cyclic peptide by treatment with dithioerythritol in basic solution. In the linear analogs, replacement of both Lys7 and Gln10 with a cysteine residue resulted in an over 100-fold loss of the biological activity when compared with the native pheromone. The corresponding cyclic disulfides were 5-10-fold more active than their sulfhydryl-containing homologs, and cyclo7,10[Cys7,L-Ala9,Cys10,Nle12] alpha-factor was 50-fold more potent than linear analogs containing Ser or Cys(CH3) in positions 7 and 10. Binding competition studies indicated that all analogs had low affinity for the alpha-factor receptor and there was a poor correlation between binding and activity in a growth arrest assay. A cyclic analog in which residues 8 and 9 were replaced by 5-aminopentanoic acid was not biologically active. Based on NMR studies, all cyclic peptides have a higher tendency to form beta-turns spanning residues 7-10 than their less active linear counterparts. The results provide strong evidence that this beta-turn is important for optimal signal transduction by alpha-factor" |
| Keywords: | "Amino Acid Sequence Binding, Competitive Chromatography, High Pressure Liquid Cystine/chemistry Disulfides/*chemistry Magnetic Resonance Spectroscopy Mating Factor Molecular Sequence Data Peptides/*chemistry/metabolism/pharmacology Pheromones/*chemistry/m;" |
| Notes: | "MedlineXue, C B McKinney, A Lu, H F Jiang, Y Becker, J M Naider, F eng GM-22087/GM/NIGMS NIH HHS/ GM22086/GM/NIGMS NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Denmark 1996/03/01 Int J Pept Protein Res. 1996 Mar; 47(3):131-41. doi: 10.1111/j.1399-3011.1996.tb01336.x" |
