Title:
LIGAND FLUORESCENT SENSOR PROTEIN AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2017/094885
Kind Code:
A1
Abstract:
The present invention involves a ligand fluorescent sensor protein that specifically responds to ligands, leading to a change in fluorescence properties. The ligand fluorescent sensor protein includes a first fluorescent protein domain, an N terminal-side linker, a ligand-bonding domain, a C terminal-side linker, and a second fluorescent protein domain. The fluorescent protein used in the ligand fluorescent sensor protein has a β-barrel structure. The first fluorescent protein domain includes a β1–β3 β-sheet region from the N-terminal of the fluorescent protein, an α-helix region continuing therefrom, and a β4–β6 β-sheet region. The second fluorescent protein domain includes a β7–β11 β-sheet region of the same fluorescent protein as the first fluorescent protein domain. The N-terminal-side linker and the C-terminal-side linker are each independently a polypeptide comprising one or a plurality of amino acids.
Inventors:
TSUBOI TAKASHI (JP)
KITAGUCHI TETSUYA (SG)
ARAI SATOSHI (SG)
UEDA HIROSHI (JP)
KITAGUCHI TETSUYA (SG)
ARAI SATOSHI (SG)
UEDA HIROSHI (JP)
Application Number:
PCT/JP2016/085902
Publication Date:
June 08, 2017
Filing Date:
December 02, 2016
Export Citation:
Assignee:
UNIV TOKYO (JP)
UNIV WASEDA (JP)
UNIV WASEDA (JP)
International Classes:
C12N15/09; C07K14/435; C07K17/00; C12N5/10; C12Q1/02; G01N33/50
Domestic Patent References:
| WO2015060430A1 | 2015-04-30 | |||
| WO2015108102A1 | 2015-07-23 |
Other References:
BAIRD, GEOFFREY ET AL.: "Circular permutation and receptor insertion within green fluorescent proteins", PROC. NATL. ACAD. SCI. USA, vol. 96, September 1999 (1999-09-01), pages 11241 - 11246, XP002201852
KITAGUCHI, TETSUYA ET AL.: "Extracellular calucium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells", BIOCHEM. J., vol. 450, 2013, pages 365 - 373, XP055376360, DOI: doi:10.1042/BJ20121022
ODAKA, HARUKI ET AL.: "Genetically-Encoded Yellow Fluorescent cAMP Indicator with an Expanded Dynamic Range for Dual-Color Imaging", PLOS ONE, vol. 9, no. 6, June 2014 (2014-06-01), pages e100252, XP055599359, DOI: 10.1371/journal.pone.0100252
GFP G-C A MP, vol. 19, no. 2, 2002, pages 135 - 145
WANG, JING, W. ET AL.: "Two-Photon Calcium Imaging Reveals an Odor-Evoked Map of Activity in the Fly Brain", CELL, vol. 112, no. 2, 24 January 2003 (2003-01-24), pages 271 - 282, XP055599364, ISSN: 0092-8674, DOI: 10.1016/S0092-8674(03)00004-7
TANIMURA, AKIHIKO: "Development and application of fluorescent protein-based indicators for live cell imaging", JOURNAL OF ORAL BIOSCIENCES, vol. 57, no. 2, 7 April 2015 (2015-04-07), pages 54 - 60, XP055599366
BISWAS, KABIR ET AL.: "The GAF DOMAIN of the cGMP-Binding, cGMP-Specific Phosphodiesterase (PDE5) Is a Sensor and a Sink for cGMP", BIOCHEMISTRY, vol. 47, no. 11, 2008, pages 3534 - 3543, XP055599369, ISSN: 0006-2960, DOI: 10.1021/bi702025w
KITAGUCHI, TETSUYA ET AL.: "Extracellular calucium influx activates adenylate cyclase 1 and potentiates insulin secretion in MIN6 cells", BIOCHEM. J., vol. 450, 2013, pages 365 - 373, XP055376360, DOI: doi:10.1042/BJ20121022
ODAKA, HARUKI ET AL.: "Genetically-Encoded Yellow Fluorescent cAMP Indicator with an Expanded Dynamic Range for Dual-Color Imaging", PLOS ONE, vol. 9, no. 6, June 2014 (2014-06-01), pages e100252, XP055599359, DOI: 10.1371/journal.pone.0100252
GFP G-C A MP, vol. 19, no. 2, 2002, pages 135 - 145
WANG, JING, W. ET AL.: "Two-Photon Calcium Imaging Reveals an Odor-Evoked Map of Activity in the Fly Brain", CELL, vol. 112, no. 2, 24 January 2003 (2003-01-24), pages 271 - 282, XP055599364, ISSN: 0092-8674, DOI: 10.1016/S0092-8674(03)00004-7
TANIMURA, AKIHIKO: "Development and application of fluorescent protein-based indicators for live cell imaging", JOURNAL OF ORAL BIOSCIENCES, vol. 57, no. 2, 7 April 2015 (2015-04-07), pages 54 - 60, XP055599366
BISWAS, KABIR ET AL.: "The GAF DOMAIN of the cGMP-Binding, cGMP-Specific Phosphodiesterase (PDE5) Is a Sensor and a Sink for cGMP", BIOCHEMISTRY, vol. 47, no. 11, 2008, pages 3534 - 3543, XP055599369, ISSN: 0006-2960, DOI: 10.1021/bi702025w
Attorney, Agent or Firm:
TANAI Sumio et al. (JP)
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