Biophysical Analysis of Heat-Induced Structural Maturation of Glutamate Dehydrogenase from a Hyperthermophilic Archaeon (ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)

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概要

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• Tertiary structure of the recombinant glutamate dehydrogenase from Thermococcus kodakaraensis KOD1 (Tk-rGDH) converts into an intact form induced by the heat treatment. This phenomenon, heat-induced structural maturation, means that high temperature plays an important role in the proper folding and oligomerization of Tk-rGDH. In this work, we analyzed the heat-induced structural maturation of Tk-rGDH by differential scanning microcalorimetry (DSC), circular dichroism (CD), and activity measurements. In DSC measurements, the peak of adsorption of non-heated Tk-rGDH (nh-Tk-rGDH) was two times smaller than that of Tk-rGDH heated at 70℃ for 30 min (h-Tk-rGDH). The transition temperature (T_m) of h-Tk-rGDH was 115℃, which was about 3℃ higher than that of nh-Tk-rGDH. In the presence of 0.5 M NaCl, the nh-Tk-rGDH showed two peaks at 107℃ and 114℃, while the h-Tk-rGDH showed a single peak at 115.7℃. The heat-induced conformational change process was monitored by changes in CD intensity at 222 nm, and the result showed that heat-induced structural maturation is irreversible. The heat treatment at 70℃ showed the highest enhancement in activity, which was 15% larger than that of heat-treated Tk-rGDH at 40℃. The results indicate that heat-induced structural maturation involves an irreversible process, transforming the non-heated form to the stable and active form.

• 社団法人日本生物工学会の論文
• 2004-05-25

著者

• Takagi Masahiro

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• Takagi M

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• Takagi Mutsumi

  International Center For Biotechnology Osaka University:(present Address)division Of Biotechnology A

• Takagi M

  Japan Advanced Inst. Of Sci. And Technol. Ishikawa Jpn

• IMANAKA Tadayuki

  Department of Synthetic and Biological Chemistry, Graduate School of Engineering, Kyoto University

• Fujiwara S

  School Of Science And Engineering Department Of Life Science Kwansei-gakuin University

• Fujiwara Shinsuke

  Department Of Biotechnology Graduate School Of Engineering Osaka University

• IZUMIKAWA NAOSHIGE

  School of Materials Science, Japan Advanced Institute of Science and Technology

• SHIRAKI KENTARO

  School of Materials Science, Japan Advanced Institute of Science and Technology

• NISHIKORI SHINGO

  School of Materials Science, Japan Advanced Institute of Science and Technology

• FUJIWARA SHINSUKE

  School of Science and Engineering, Departmeng of Life Science, Kwansei-Gakuin University

• Takagi M

  Nagasaki Univ. Nagasaki Jpn

• Imanaka Tadayuki

  Department Of Applied Biotechnology Faculty Of Engineering Osaka University

• Imanaka Tadayuki

  Department Of Biotechnology Faculty Of Eigineering Osaka University

• Imanaka Tadayuki

  Department Of Synthetic And Biological Chemistry Graduate School Of Engineering Kyoto University

• Imanaka T

  Department Of Biotechnology Faculty Of Engineering Osaka University

• Imanaka Tadayuki

  Department Of Synthetic Chemistry And Biological Chemistry Graduate School Of Engineering.kyoto Univ

• Shiraki K

  Institute Of Applied Physics University Of Tsukuba

• Fujiwara Shinsuke

  School Of Science And Engineering Department Of Life Science Kwansei-gakuin University

• Takagi Mutsumi

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• Takagi Masahiro

  School Of Material Science Japan Advanced Institute Of Science And Technology

• Imanaka T

  Dep. Of Biotechnology Coll. Of Life Sciences Ritsumeikan Univ.

• Izumikawa Naoshige

  School Of Materials Science Japan Advanced Institute Of Science And Technology

• Nishikori Shingo

  Division Of Molecular Cell Biology Institute Of Molecular Embryology And Genetics Kumamoto Universit

• Takagi Masahiro

  Graduate School Of Biological Sciences Nara Institute Of Science And Technology (naist)

• Nishikori Shingo

  School of Materials Science, Japan Advanced Institute for Science and Technology

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