Amino Acids Conserved at the C-Terminal Half of the Ribonuclease T2 Family Contribute to Protein Stability of the Enzymes
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概要
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The ribonuclease MC1 (RNase MC1) from the seeds of the bitter gourd belongs to the RNase T2 family. We evaluated the contribution of 11 amino acids conserved in the RNase T2 family to protein folding of RNase MC1. Thermal unfolding experiments showed that substitution of Tyr101, Phe102, Ala105, and Phe190 resulted in a significant decrease in themostability; the Tm values were 47–58 °C compared to that for the wild type (64 °C). Mutations of Pro125, Gly127, Gly144, and Val165 caused a moderate decrease in thermostability (Tm: 60–62 °C). In contrast, mutations of Asp107 and Gly173 did little effect on thermostability. The contribution of Tyr101, Phe102, Pro125, and Gly127 to protein stability was further corroborated by means of Gdn–HCl unfolding and protease digestions. Taken together, it appeared that Tyr101, Phe102, Ala105, Pro125, Gly127, Gly144, Leu162, Val165, and Phe190 conserved in the RNase T2 family play an important role in the stability of the proteins.
- 社団法人 日本農芸化学会の論文
- 2004-08-23
著者
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KIMURA Makoto
Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University
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KAKUTA Yoshimitsu
Laboratory of Structural Biology, Graduate School of Systems Life Sciences, Kyushu University
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Numata Tomoyuki
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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KIMURA Kazumi
Laboratory of Biochemistry, Department of Bioscience and Biotechnology, Faculty of Agriculture, Grad
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Kimura Kazumi
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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Numata Tomoyuki
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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Kakuta Yoshimitsu
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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Kimura Makoto
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Graduate School Faculty Of Agr
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Kimura Makoto
Laboratory Of Biochemistry Department Of Agricultural Chemistry Faculty And Graduate School Of Agric
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Kimura Makoto
Laboratory Of Biochemistry Department Of Bioscience And Biotechnology Faculty Of Agriculture Graduat
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KIMURA Makoto
Laboratory for Remediation Research, Plant Science Center, RIKEN
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