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Redox and metal-regulated oligomeric state for human porphobilinogen synthase activation

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Abstract

The oligomeric state of human porphobilinogen synthase (PBGS) [EC.4.2.1.24] is homooctamer, which consists of conformationally heterogenous subunits in the tertiary structure under air-saturated conditions. When PBGS is activated by reducing agent with zinc ion, a reservoir zinc ion coordinated by Cys223 is transferred in the active center to be coordinated by Cys122, Cys124, and Cys132 (Sawada et al. in J Biol Inorg Chem 10:199–207, 2005). The latter zinc ion serves as an electrophilic catalysis. In this study, we investigated a conformational change associated with the PBGS activation by reducing agent and zinc ion using analytical ultracentrifugation, negative staining electron microscopy, native PAGE, and enzyme activity staining. The results are in good agreement with our notion that the main component of PBGS is octamer with a few percent of hexamer and that the octamer changes spatial subunit arrangement upon reduction and further addition of zinc ion, accompanying decrease in f/f 0. It is concluded that redox-regulated PBGS activation via cleavage of disulfide bonds among Cys122, Cys124, and Cys132 and coordination with zinc ion is closely linked to change in the oligomeric state.

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Authors and Affiliations

  1. Department of Environmental Medicine, Nippon Medical School, 1-1-5 Sendagi Bunkyo-ku, Tokyo, 113-8602, Japan

    N. Sawada, N. Nagahara & M. Minami

  2. Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259-B39, Nagatsuta-cho, Midori-ku, Yokohama, 226-8510, Japan

    F. Arisaka

  3. Biomedicinal Information Research Center, National Institute of Advanced Industrial Science and Technology, 2-41-6, Aomi, Koto-ku, Tokyo, 135-0064, Japan

    K. Mitsuoka

Authors
  1. N. Sawada
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  2. N. Nagahara
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  3. F. Arisaka
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  4. K. Mitsuoka
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  5. M. Minami
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Corresponding author

Correspondence to N. Nagahara.

Additional information

N. Sawada and N. Nagahara contributed equally to the results of this article.

This article is published as part of the Special Issue on Sulfur- and seleno-containing amino acids.

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Sawada, N., Nagahara, N., Arisaka, F. et al. Redox and metal-regulated oligomeric state for human porphobilinogen synthase activation. Amino Acids 41, 173–180 (2011). https://doi.org/10.1007/s00726-010-0570-y

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  • DOI: https://doi.org/10.1007/s00726-010-0570-y

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