| Title: | Probing the binding domain of the Saccharomyces cerevisiae alpha-mating factor receptor with rluorescent ligands |
| Author(s): | Ding FX; Lee BK; Hauser M; Davenport L; Becker JM; Naider F; |
| Address: | "Department of Chemistry, The College of Staten Island of the City University of New York, Staten Island, New York 10314, USA" |
| ISSN/ISBN: | 0006-2960 (Print) 0006-2960 (Linking) |
| Abstract: | "Three analogues of the alpha-mating factor pheromone of Saccharomyces cerevisiae containing the 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) group were synthesized that had high binding affinity to the receptor and retained biological activity. The fluorescence emission maximum of the NBD group in [K7(NBD),Nle(12)]-alpha-factor was blue shifted by 35 nm compared to buffer when the pheromone bound to its receptor. Fluorescence quenching experiments revealed that the NBD group in [K7(NBD),Nle(12)]-alpha-factor bound to the receptor was shielded from collision with iodide anion when in aqueous buffer. In contrast, the emission maximum of NBD in [K7(ahNBD),Nle(12)]-alpha-factor or [Orn7(NBD),Nle(12)]-alpha-factor was not significantly shifted and iodide anion efficiently quenched the fluorescence of these derivatives when they were bound to receptor. The fluorescence investigation suggests that when the alpha-factor is bound to its receptor, K7 resides in an environment that has both hydrophobic and hydrophilic groups within a few angstroms of each other" |
| Keywords: | "4-Chloro-7-nitrobenzofurazan/chemical synthesis/metabolism Binding, Competitive Cell Membrane/metabolism Fluorescent Dyes/chemical synthesis/*metabolism Fungal Proteins/*metabolism Ligands Mating Factor Peptides/*metabolism Receptors, Mating Factor Recept;" |
| Notes: | "MedlineDing, F X Lee, B K Hauser, M Davenport, L Becker, J M Naider, F eng GM22086/GM/NIGMS NIH HHS/ GM22087/GM/NIGMS NIH HHS/ Research Support, U.S. Gov't, P.H.S. 2001/02/15 Biochemistry. 2001 Jan 30; 40(4):1102-8. doi: 10.1021/bi0021535" |
