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Molecular characterization of the group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3

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Abstract

The group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3 (PhCPN) and its functional cooperation with the cognate prefoldin were investigated. PhCPN existed as a homo-oligomer in a double-ring structure, which protected the citrate synthase of a porcine heart from thermal aggregation at 45°C, and did the same on the isopropylmalate dehydrogenase (IPMDH) of a thermophilic bacterium, Thermus thermophilus HB8, at 90°C. PhCPN also enhanced the refolding of green fluorescent protein (GFP), which had been unfolded by low pH, in an ATP-dependent manner. Unexpectedly, functional cooperation between PhCPN and Pyrococcus prefoldin (PhPFD) in the refolding of GFP was not observed. Instead, cooperation between PhCPN and PhPFD was observed in the refolding of IPMDH unfolded with guanidine hydrochloride. Although PhCPN alone was not effective in the refolding of IPMDH, the refolding efficiency was enhanced by the cooperation of PhCPN with PhPFD.

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Acknowledgements

The work reported here is a part of the 21st Century COE (Center of Excellence) program of “Future Nano-Materials” research and education project, which is financially supported by the Ministry of Education, Science, Sports, Culture, and Technology through Tokyo University of Agriculture and Technology. This work was also supported by grants-in-aid for scientific research on priority areas (13033008 14037216, and 15032212), and a grant of the National Project on Protein Structural and Functional Analyses from the Ministry of Education, Science, Sports and Culture of Japan to M.Y. We appreciate Dr. Kawarabayasi for giving a shotgun clone of P. horikoshii OT3 containing a chaperonin ORF, PH0017.

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Authors and Affiliations

  1. Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan

    Mina Okochi, Hiroki Matsuzaki, Tomoko Nomura & Masafumi Yohda

  2. Biological Information Research Center, National Institute of Advanced Industrial Science and Technology, Tsukuba Central-6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan

    Noriyuki Ishii

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  1. Mina Okochi
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  2. Hiroki Matsuzaki
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  4. Noriyuki Ishii
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  5. Masafumi Yohda
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Correspondence to Masafumi Yohda.

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Communicated by F. Robb

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Okochi, M., Matsuzaki, H., Nomura, T. et al. Molecular characterization of the group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3. Extremophiles 9, 127–134 (2005). https://doi.org/10.1007/s00792-004-0427-y

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  • DOI: https://doi.org/10.1007/s00792-004-0427-y

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