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A Demetallation Method for IMP-1 Metallo-β-Lactamase with Restored Enzymatic Activity Upon Addition of Metal Ion(s)

Authors

  • Prof. Dr. Yoshihiro Yamaguchi,

    Corresponding author
    1. Environmental Safety Center, Kumamoto University, 39-1 Kurokami 2-Chome, Kumamoto 860-8555 (Japan)
    • Environmental Safety Center, Kumamoto University, 39-1 Kurokami 2-Chome, Kumamoto 860-8555 (Japan)
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  • Prof. Dr. Shijia Ding,

    1. Department of Medical Laboratory, Chongqing University of Medical Sciences, Chongqing 400016 (China)
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  • Emi Murakami,

    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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  • Kayo Imamura,

    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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  • Sachiko Fuchigami,

    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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  • Ryo Hashiguchi,

    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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  • Prof. Dr. Katsuhide Yutani,

    1. RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo, Hyogo 679-5148 (Japan)
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  • Dr. Hiromasa Mori,

    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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  • Prof. Dr. Shinnichiro Suzuki,

    1. Department of Chemistry, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043 (Japan)
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  • Prof. Dr. Yoshichika Arakawa,

    1. Department of Bacteriology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550 (Japan)
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  • Prof. Dr. Hiromasa Kurosaki

    Corresponding author
    1. Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
    • Department of Structure–Function Physical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmcahi, Kumamoto 862-0973 (Japan)
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Abstract

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Think zinc: We have developed a new preparation method for generating the apo-enzyme of IMP-1 metallo-β-lactamase (apo-IMP-1) with the combination of a chelating agent (EDTA) and desalting column chromatography. An advantage of this method is that enzymatic activity of the apo-enzyme comparable to that observed with the native enzyme could be restored upon addition of ZnII ions (see scheme).

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