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Construction of streptavidin-luciferase fusion protein for ATP sensing with fixed form

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Abstract

A fusion protein consisting of streptavidin and firefly luciferase was constructed to establish an accurate measuring technique of local ATP concentration. The fusion protein retained the binding ability of streptavidin and enzymatic activity of luciferase. Also, it could detect the concentration of antigens and could determine nanomolar concentrations of ATP in its fixed form via interactions with biotin-conjugated antibodies.

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References

  • Arcuino G, Lin JH-C, Takano T, Liu C, Jiang L, Gao Q, Kang J, Nedergaard M (2002) Intercellular calcium signaling mediated by point-source burst release of ATP. Proc. Natl. Acad. Sci. USA 99: 9840–9845.

    PubMed  Google Scholar 

  • Askenasy N, Yolcu ES, Wang Z, Shirwan H (2003) Display of fas ligand protein on cardiac vasculature as a novel means of regulating allograft rejection. Circulation 107: 525–1531.

    Google Scholar 

  • Beigi R, Kobatake E, Aizawa M, Dubyak GR (1999) Detection of local ATP release from activated platelets using cell surfaceattached firefly luciferase. Am. J. Physiol. Cell. Physiol. 276: C267–C278.

    Google Scholar 

  • Deluca M, McElroyWD (1978) Purification and properties of firefly luciferase. Meth. Enzymol. 57: 3–15.

    Google Scholar 

  • Ebihara T, Takayama H, Yanagida Y, Kobatake E, Aizawa M (2002) Thermostabilization of protein A-luciferase fusion protein by single amino acid mutation. Biotechnol. Lett. 24: 147–149.

    Google Scholar 

  • Funabashi H, Imajo T, Kojima J, Kobatake E, Aizawa M (1999) Bioluminescent monitoring of intracellular ATP during fermentation. Luminescence 14: 291–296.

    PubMed  Google Scholar 

  • Huang R-P, Huang R, Fan Y, Lin Y (2001) Simultaneous detection of multiple cytokines from conditioned media and patient's sera by an antibody-based protein array system. Anal. Biochem. 294: 55–62.

    PubMed  Google Scholar 

  • Lundin A (2000) Use of firefly luciferase in ATP-related assays of biomass, enzymes, and metabolites. Meth. Enzymol. 305: 346–370.

    Google Scholar 

  • Nishihara K, Kanemori M, Kitagawa M, Yanagi H, Yura T (1998) Chaperone coexpression plasmids: differential and synergistic roles of DnaK-DnaJ-GrpE and GroEL-GroES in assisting folding of an allergen of Japanese cedar pollen, Cryj2, in Escherichia coli. Appl. Env. Microbiol. 64: 1694–1699.

    Google Scholar 

  • Ohno K, Levin B, Meruelo D (1996) Cell-specific, multidrug delivery system using streptavidin-protein A fusion protein. Biochem. Mol. Med. 58: 227–233.

    PubMed  Google Scholar 

  • White PJ, Squirrel DJ, Arnaud P, Lowe CR, Murray JAH (1996) Improved thermostability of the North American firefly luciferase: saturation mutagenesis at position 354. Biochem. J. 319: 343–350.

    PubMed  Google Scholar 

  • Wilchek M, Bayer EA (1990) Avidin-biotin technology. Meth. Enzymol. 184: 1–671.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan

    Makiko Nakamura, Masayasu Mie, Hisakage Funabashi & Eiry Kobatake

Authors
  1. Makiko Nakamura
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  2. Masayasu Mie
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  3. Hisakage Funabashi
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  4. Eiry Kobatake
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Nakamura, M., Mie, M., Funabashi, H. et al. Construction of streptavidin-luciferase fusion protein for ATP sensing with fixed form. Biotechnology Letters 26, 1061–1066 (2004). https://doi.org/10.1023/B:BILE.0000032966.17759.08

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  • DOI: https://doi.org/10.1023/B:BILE.0000032966.17759.08

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