Title: | Development of a Highly Active Fluorescence-Based Detector for Yeast G Protein-Coupled Receptor Ste2p |
Author(s): | Hong JW; Ahn HJ; Baek JS; Hong EY; Jin DH; Khang YH; Hong NJ; |
Address: | "School of Biotechnology, Yeungnam University, Gyeongsan 38541, Republic of Korea. Asan Institute for Life Science, Department of Convergence Medicine, College of Medicine, University of Ulsan 44610, Republic of Korea" |
ISSN/ISBN: | 1738-8872 (Electronic) 1017-7825 (Linking) |
Abstract: | "Twenty analogs of [Orn(6),D-Ala(9)]alpha-factor were synthesized and assayed for their biological activities: seven analogs of [Orn(6),X(9)]alpha-factor, seven analogs of [X(6),D-Ala(9)]alpha-factor, five analogs of [X(5),X(6),D-Ala(9)]alpha-factor, and native alpha-factor (X = amino acids). Their biological activities (halo, gene induction, and affinity) were measured using S. cerevisiae Y7925 and LM102 and compared with those of native alpha-factor (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. [Dap(6),D-Ala(9)]alpha-factor with weak halo activity (10%) showed the highest receptor affinity (??? 230%) and the highest gene induction activity (167%). [Arg(6),D-Ala(9)]alpha-factor showed the highest halo activity (2,000%). The number of active binding sites per cell (about 20,000 for strain LM102) was determined using a newly-designed fluorescence-based detector, [Arg(6),D-Ala(9)]alpha-factor-Edan, with high sensitivity (12,500-fold higher than the absorption-based detector [Orn(6)]alpha-factor-[Cys](3))" |
Keywords: | "Amino Acid Sequence Binding Sites Binding, Competitive Fluorescence Gene Expression Genes, Reporter/genetics Mating Factor/*analysis/chemical synthesis/chemistry/*metabolism Protein Binding Receptors, G-Protein-Coupled/genetics/*metabolism Receptors, Mati;" |
Notes: | "MedlineHong, Jin Woo Ahn, Hee Jun Baek, Jee Su Hong, Eun Young Jin, Dong Hoon Khang, Yong Ho Hong, Nam Joo eng Korea (South) 2018/11/18 J Microbiol Biotechnol. 2018 Oct 28; 28(10):1589-1603. doi: 10.4014/jmb.1805.04045" |