본문 바로가기
KERD
논문KCI 등재

Development of a Highly Active Fluorescence-Based Detector for Yeast G Protein-Coupled Receptor Ste2p

1

조회

0

다운로드

0

인용

0

참고문헌

키워드

주제분류생물학

초록

Twenty analogs of [Orn6,D-Ala9]α-factor were synthesized and assayed for their biological activities: seven analogs of [Orn6,X9]α-factor, seven analogs of [X6,D-Ala9]α-factor, five analogs of [X5,X6,D-Ala9]α-factor, and native α-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 α-factor (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. [Dap6,D-Ala9]α-factor with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). [Arg6,D-Ala9]α-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 newlydesigned fluorescence-based detector, [Arg6,D-Ala9]α-factor-Edan, with high sensitivity (12,500-fold higher than the absorption-based detector [Orn6]α-factor-[Cys]3).

Abstract

Twenty analogs of [Orn6,D-Ala9]α-factor were synthesized and assayed for their biological activities: seven analogs of [Orn6,X9]α-factor, seven analogs of [X6,D-Ala9]α-factor, five analogs of [X5,X6,D-Ala9]α-factor, and native α-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 α-factor (100%). G protein-coupled receptor was expressed in strain LM102 containing pESC-LEU-STE2 vector. [Dap6,D-Ala9]α-factor with weak halo activity (10%) showed the highest receptor affinity (> 230%) and the highest gene induction activity (167%). [Arg6,D-Ala9]α-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 newlydesigned fluorescence-based detector, [Arg6,D-Ala9]α-factor-Edan, with high sensitivity (12,500-fold higher than the absorption-based detector [Orn6]α-factor-[Cys]3).

참고문헌 (39)

  1. Dohlamn HG., "G proteins and pheromone signaling", Annu. Rev. Physiol, 64, pp.129~152 (2002)
  2. Schöneberg T, "Mutant G-protein-coupled receptors as a cause of human diseases", Pharmac. Ther, 104, pp.173~206 (2004)
  3. Klabunde T, "Drug design strategies for targeting G-protein-coupled receptors", Chembiochem, 3, pp.928~944 (2002)
  4. Naider F, "The alpha-factor mating pheromone of Saccharomyces cerevisiae : a model for studying the interaction of peptide hormones and G protein-coupled receptors", Peptides, 25, pp.1441~1463 (2004)
  5. Stotzler D, "Isolation and characterization of four related peptides exhibiting alpha factor activity from Saccharomyces cerevisiae", Eur. J. Biochem, 65, pp.257~262 (1976)
  6. Naider F, "Structure-activity relationships of the yeast alpha-factor", CRC Crit. Rev. Biochem, 21, pp.225~248 (1986)
  7. Garbow JR, "Conformational analysis of the Saccharomyces cerevisiae tridecapeptide mating pheromone by 13C, 15N rotational-echo double resonance nuclear magnetic resonance spectroscopy", Biochemistry, 33, pp.10094~10099 (1994)
  8. Henry LK, "Identification of a contact region between the tridecapeptide alpha-factor mating pheromone of Saccharomyces cerevisiae and its G protein-coupled receptor by photoaffinity labeling", Biochemistry, 41, pp.6128~6139 (2002)
  9. Ding FX, "Probing the binding domain of the Saccharomyces cerevisiae alpha-mating factor receptor with rluorescent ligands", Biochemistry, 40, pp.1102~1108 (2001)
  10. Abel MG, "Structure-function analysis of the Saccharomyces cerevisiae tridecapeptide pheromone using alanine-scanned analogs", J. Pept. Res, 52, pp.95~106 (1998)
  11. alpha-factor in solution and in the presence of lipid, "Biochemistry", 1993, 908~917
  12. Hee Jun Ahn, "Highly Active Analogs of α-Factor and Their Activities Against Saccharomyces cerevisiae", Bulletin of the Korean Chemical Society, 35(5), pp.1365~1374 (2014), 대한화학회[DOI]
  13. 안희준, "Spectrophotometric Determination of Affinities of α-Factors for Their G Protein-Coupled Receptors in Saccharomyces cerevisiae", Bulletin of the Korean Chemical Society, 36(7), pp.1885~1896 (2015), 대한화학회[DOI]
  14. Kim KM, "Multiple regulatory roles of the carboxy terminus of Ste2p a yeast GPCR", Pharmacol. Res, 65, pp.31~40 (2012)
  15. Hanahan D., "Studies on transformation of Escherichia coli with plasmids", J. Mol. Biol, 166, pp.557~580 (1983)
  16. Gietz RD, "High-efficiency yeast transformation using the LiAc", SS carrier DNA, Nat. Protoc, pp.2007, 2[DOI]
  17. 홍남주, "Structure-Activity Relationships of 13- and 14-Membered Cyclic Partial Retro-Inverso Pentapeptides Related to Enkephalin", Bulletin of the Korean Chemical Society, 31(4), pp.874~880 (2010), 대한화학회
  18. Dong Hee Kim, "Activity Profiles of Linear, Cyclic Monomer and Cyclic Dimer of Enkephalin", Bulletin of the Korean Chemical Society, 33(1), pp.261~269 (2012), 대한화학회
  19. Li J, "Green fluorescent protein in Saccharomyces cerevisiae: real-time studies of the GAL1promoter", Biotechnol. Bioeng, 70, pp.187~196 (2000)
  20. Taylor RG, "E. coli host strains significantly affect the quality of small scale plasmid DNA preparations used for sequencing", Nucleic Acids Res, 21, pp.1677~1678 (1993)
  21. Bimboim HC, "A rapid alkaline extraction procedure for screening recombinant plasmid DNA", Nucleic Acid Res, 7, pp.1513~1523 (1979)
  22. Chrambach A, "Polyacrylamide gel electrophoresis", Science, 172, pp.440~451 (1971)
  23. Thibodeau SA, "High-throughput β-galactosidase assay for bacterial cell-based reporter systems", Biotechniques, 36, pp.410~415 (2004)
  24. Vidal-Aroca, "One-step high-throughput assay for quantitative detection of beta-galactosidase activity in intact gram-negative bacteria, yeast, and mammalian cells", Biotechniques, 40, pp.433~434 (2006)
  25. Plovins A, "Use of fluorescein-di-beta-D-galactopyranoside (FDG)and C12-FDG as substrates for beta-galactosidase detection by flow cytometry in animal, bacterial, and yeast cells", Appl. Environ. Microbiol, 60, pp.4638~4641 (1994)
  26. Raths SK, "Peptide Analogues Compete with the Binding of α-factor to Its Receptor in Saccharomyces cerevisiae", J. Biol. Chem, 263, pp.17333~17341 (1988)
  27. Kippert F., "A rapid permeabilization procedure for accurate quantitative determination of β-galactosidase activity in yeast cells", FEMS Microbiol Lett, 128, pp.201~206 (1995)
  28. Jenness DD, "Down regulation of the α-factor pheromone receptor in S. cerevisiae", Cell, 46, pp.345~353 (1986)
  29. Mathew E, "Differential interactions of fluorescent agonists and antagonists with the yeast G protein coupled receptor Ste2p", J. Mol. Biol, 409, pp.513~528 (2011)
  30. Son CD, "Identification of ligand binding regions of the Saccharomyces cerevisiae α-factor pheromone receptor by photoaffinity cross-linking", Biochemistry, 43, pp.13193~13203 (2004)
  31. Xue CB, "Probing the functional conformation of the tridecapeptide mating pheromone of Saccharomyces cerevisiae through study of disulfide-constrained analogs", Int. J. Pept. Protein. Res, 47, pp.131~141 (1996)
  32. Banerjee R, "Conformational preferences of a short Aib", Ala-based water-soluble peptide as a function of temperature, 2009, pp.76, 184~200
  33. Wakamatsu K, "Conformations of yeast α-mating factor and analog peptides as bound to phospholipid bilayer", Eur. J. Biochem, 163, pp.331~338 (1987)
  34. Higashijima T, "Conformations of tridecapeptide α-mating factor from yeast Saccharomyces cerevisiae and analog peptides in aqueous solution", Eur. J. Biochem, 140, pp.163~171 (1984)
  35. Marsh L., "Substitutions in the hydrophobic core of the alpha-factor receptor of Saccharomyces cerevisiae permit response to Saccharomyces kluyveri alpha-factor and to antagonist", Mol. Cell. Biol, 12, pp.3959~3966 (1992)
  36. Liu S, "Position 13 analogs of the tridecapeptide mating pheromone from Saccharomyces cerevisiae: design of an iodinatable ligand for receptor binding", J. Pept. Res, 56, pp.24~34 (2000)
  37. Widmann C, "Mitogen-activated protein kinase: conservation of a threekinase module from yeast to human", Physiol. Rev, 79, pp.143~180 (1999)
  38. Oehlen LJ, "Ste12 and Mcm1regulate cell cycle-dependent transcription of FAR1", Mol. Cell. Biol, 16, pp.2830~2837 (1996)
  39. Bardwell L, "Signaling in the yeast pheromone response pathway: specific and high-affinity interaction of the mitogen-activated protein (MAP) kinases Kss1 and FUS3 with the upstream MAP kinase kinase Ste7", Mol. Cell. Biol, 16, pp.3637~3650 (1996)

관련 논문AI추천

Probing spatiotemporally organized GPCR signaling using genetically encoded molecular toolsKwon Yonghoon, Mehta Sohum, Zhang Jin생화학분자생물학회 · Experimental and Molecular Medicine2025 · 제57권 Global Transcriptomic Analysis Identifies Gene Expression Programs Regulated by α-Factor in YeastLee Jung-Shin, Yeom Soojin한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2025 · 제35권 Signal Transduction of C-Terminal Phosphorylation Regions for Equine Luteinizing Hormone/Chorionic Gonadotropin Receptor (eLH/CGR)민관식한국발생생물학회 · 발생과 생식2022 · 제26권 제1호Novel Bombesin Analogues Conjugated with DOTA-Ala(SO3H)-4 aminobenzoic acid and DOTA-Lys(glucose)-4 aminobenzoic acid: Synthesis, Radiolabeling, and Gastrin Releasing Peptide Receptor Binding Affinity김진주, 임재청, 조은하, 최상무(사)한국방사선산업학회 · 방사선산업학회지2013 · 제7권 제2호Modulation of Kex2p Cleavage Site for In Vitro Processing of Recombinant Proteins Produced by Saccharomyces cerevisiaeBae Jung-Hoon, Kim Mi-Jin, Kim Seung Hwa, Park Hyun-Joo, Park Se-Lin, Sohn Jung-Hoon, Sung Bong Hyun한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2023 · 제33권 제11호The C-terminal Phosphorylation Sites of eel Follicle-Stimulating Hormone Receptor are Important Role in the Signal Transduction강명화, 민관식, Jeong-Min Kim, Munkhzaya Byambaragchaa한국발생생물학회 · 발생과 생식2018 · 제22권 제2호

이 간행물의 다른 논문 (12건)