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J Microbiol Biotechnol


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"
Journal Title:J Microbiol Biotechnol
Year:2018
Volume:28
Issue:10
Page Number:1589 - 1603
DOI: 10.4014/jmb.1805.04045
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"

 
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