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A Detection Method for GLUT4 Exocytosis Based on Spontaneous Split Luciferase Complementation

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Abstract

Glucose transporter 4 (GLUT4) is an insulin-regulated glucose transporter, which is vital for blood glucose homeostasis. To clarify the physiological roles of GLUT4, quantitative measurement of GLUT4 exocytosis is indispensable. Herein, we show a rapid detection system for GLUT4 on the cell surface using spontaneous split-luciferase reconstitution. Upon insulin-induced GLUT4 exocytosis, GLUT4 was exposed outside, where luciferase is reconstituted and emitted luminescence. Pretreatment with inhibitors reduced the insulin-induced signal elevation. The results indicate that the developed method is applicable to high-throughput analysis on GLUT4 trafficking, which will greatly accelerate comprehensive research on the physiological roles of GLUT4.

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Acknowledgements

This work was supported by CREST (JPMJCR1752 to T. O.) from the Japan Science and Technology (JST), and the Japan Society for the Promotion of Science (JSPS) KAKENHI (Grants-in-Aid for Scientific Research (S) 26220805, (A) 19H00900, and Innovative Areas JP 19H04952 to T. O.).

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Authors and Affiliations

  1. Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan

    Mizuki Endo, Masashi Miyasaki, Qiaojing Li, Genki Kawamura & Takeaki Ozawa

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  1. Mizuki Endo
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  2. Masashi Miyasaki
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  3. Qiaojing Li
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  4. Genki Kawamura
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  5. Takeaki Ozawa
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Correspondence to Takeaki Ozawa.

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Endo, M., Miyasaki, M., Li, Q. et al. A Detection Method for GLUT4 Exocytosis Based on Spontaneous Split Luciferase Complementation. ANAL. SCI. 35, 835–838 (2019). https://doi.org/10.2116/analsci.19C003

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  • DOI: https://doi.org/10.2116/analsci.19C003

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