Bright fluorescence monitoring system utilizing Zoanthus sp. green fluorescent protein (ZsGreen) for human G-protein-coupled receptor signaling in microbial yeast cells

PloS One
Yasuyuki NakamuraAkihiko Kondo

Abstract

G-protein-coupled receptors (GPCRs) are currently the most important pharmaceutical targets for drug discovery because they regulate a wide variety of physiological processes. Consequently, simple and convenient detection systems for ligands that regulate the function of GPCR have attracted attention as powerful tools for new drug development. We previously developed a yeast-based fluorescence reporter ligand detection system using flow cytometry. However, using this conventional detection system, fluorescence from a cell expressing GFP and responding to a ligand is weak, making detection of these cells by fluorescence microscopy difficult. We here report improvements to the conventional yeast fluorescence reporter assay system resulting in the development of a new highly-sensitive fluorescence reporter assay system with extremely bright fluorescence and high signal-to-noise (S/N) ratio. This new system allowed the easy detection of GPCR signaling in yeast using fluorescence microscopy. Somatostatin receptor and neurotensin receptor (implicated in Alzheimer's disease and Parkinson's disease, respectively) were chosen as human GPCR(s). The facile detection of binding to these receptors by cognate peptide ligands was demonstrated...Continue Reading

References

Mar 25, 1992·Nucleic Acids Research·D GietzR H Schiestl
Jul 31, 1984·Biochemical and Biophysical Research Communications·N MinaminoH Matsuo
Feb 1, 1995·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·T GudermannG Schultz
Mar 1, 1995·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·B NürnbergG Schultz
Jan 26, 1995·Biochemical and Biophysical Research Communications·Y XuM Berelowitz
Jan 1, 1994·The Journal of Clinical Endocrinology and Metabolism·S W LambertsJ C Reubi
Jun 1, 1997·Nature Biotechnology·E T Boder, K D Wittrup
Dec 9, 1997·Applied Microbiology and Biotechnology·T MuraiA Tanaka
Oct 3, 1999·Nature Biotechnology·M V MatzS A Lukyanov
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Jun 1, 2001·Journal of Neurology, Neurosurgery, and Psychiatry·R M SchimpffW Rostène
Jun 1, 2001·Nature Biotechnology·E MedicoP Soriano
Jan 5, 2002·Applied Microbiology and Biotechnology·Y NakamuraA Kondo
Oct 11, 2003·Biotechnology Advances·M Ueda, A Tanaka
Jan 13, 2004·Applied Microbiology and Biotechnology·A Kondo, M Ueda
Jun 1, 2005·Trends in Biotechnology·Graham LaddsJohn Davey
Jul 25, 2006·Nature Methods·Amir AharoniStephen G Withers
Sep 18, 2007·Current Opinion in Structural Biology·S Annie Gai, K Dane Wittrup
Sep 27, 2007·PloS One·Dietmar Dirnberger, Klaus Seuwen
Oct 24, 2007·Nature·Søren G F RasmussenBrian K Kobilka
Jan 15, 2008·Molecular and Cellular Endocrinology·Anat Ben-Shlomo, Shlomo Melmed
Mar 14, 2008·Combinatorial Chemistry & High Throughput Screening·Lauren R PepperEric V Shusta
Jan 14, 2009·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Seiji ShibasakiMitsuyoshi Ueda

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Citations

Apr 20, 2017·Biomedical Microdevices·Raja K VadiveluNam-Trung Nguyen
May 29, 2018·Applied and Environmental Microbiology·Yasuyuki NakamuraAkihiko Kondo
Aug 27, 2014·FEMS Yeast Research·Adebola AdeniranKeith E J Tyo
Dec 12, 2019·FEMS Yeast Research·Bettina Lengger, Michael K Jensen
May 23, 2018·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Martha Lopez-YrigoyenLesley M Forrester
Dec 19, 2020·Biochemical Pharmacology·Xuesong WangAdriaan P IJzerman

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Methods Mentioned

BETA
fluorescence
flow cytometry
fluorescence microscopy
biosensor
PCR
Assay
peptide display

Software Mentioned

BD FACSDiva
ImageJ

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