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

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  • ENDO Mizuki
    Department of Chemistry, School of Science, The University of Tokyo
  • MIYASAKI Masashi
    Department of Chemistry, School of Science, The University of Tokyo
  • LI Qiaojing
    Department of Chemistry, School of Science, The University of Tokyo
  • KAWAMURA Genki
    Department of Chemistry, School of Science, The University of Tokyo
  • OZAWA Takeaki
    Department of Chemistry, School of Science, The University of Tokyo

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Abstract

<p>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.</p>

Journal

  • Analytical Sciences

    Analytical Sciences 35 (8), 835-838, 2019-08-10

    The Japan Society for Analytical Chemistry

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