Cation-driven Optical Properties of Artificial Luciferases

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Author(s)

    • MILLER Simon MILLER Simon
    • Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)
    • SENDA Toshiya
    • Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)|Department of Materials Structure Science, School of High Energy Accelerator Science, The Graduate University of Advanced Studies (Soken-dai)
    • NISHIHARA Ryo
    • Department of Applied Chemistry, Faculty of Science and Technology, Keio University
    • SUZUKI Koji
    • Department of Applied Chemistry, Faculty of Science and Technology, Keio University

Abstract

The present study demonstrates cation-driven optical properties of artificial luciferases (ALucs) from copepod luciferases, as an optical readout for bioanalysis. An assignment of the supersecondary structure code (SSC) of ALucs revealed that ALucs carry a helix-loop-helix structure, which appears at the same sites of the EF-hands of typical Ca<sup>2+</sup>-binding proteins. A mutagenesis study shows that the EF-hand-like structure is a pivotal site for enzymatic activity. The effects of 20 kinds of mono- and multivalent cations on ALuc activities were estimated with column-purified ALuc16. High pH values boost the ALuc activities with both the native coelenterazine and an analog called 6-pi-OH-CTZ. Multivalent cations, Ca(II), Mg(II), and Cr(VI), elevate and prolong the ALuc activities, whereas Co(II), Cu(II) and Pb(II) greatly hamper the ALuc activities. Ca(II) greatly prolongs the optical intensities, suggesting a contribution to the structural robustness of ALucs. The inhibitory effect of multivalent cations on the ALuc activities was utilized for creating dose-response curves. The intrinsic cation-driven selectivity and optical intensity of ALucs enable researchers to constitute <i>de novo</i> biosensors for multivalent cations.

Journal

  • Analytical Sciences

    Analytical Sciences 31(10), 955-960, 2015

    The Japan Society for Analytical Chemistry

Codes

  • NII Article ID (NAID)
    130005102883
  • NII NACSIS-CAT ID (NCID)
    AA10500785
  • Text Lang
    ENG
  • ISSN
    0910-6340
  • NDL Article ID
    026791462
  • NDL Call No.
    Z54-F482
  • Data Source
    NDL  J-STAGE 
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