Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Biochemical and NMR Characterization of MTH1880 Mutant Proteins for Folding-Unfolding Studies

  • Kim, Hee-Youn (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Ryu, Soo-Young (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Yun, Ji-Hye (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University) ;
  • Kim, Suhk-Mann (Department of Chemistry and Chemistry Institute for Functional Materials, Pusan National University) ;
  • Chang, Ik-Soo (Creative Research Initiatives Center for Proteome Biophysics, Department of Physics, Pusan National University) ;
  • Lee, Weon-Tae (Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University)
  • Received : 2010.07.15
  • Accepted : 2010.09.16
  • Published : 2010.12.20
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Abstract

MTH1880 is a hypothetical protein derived from Methanobacterium thermoautotrophicum, thermophilic methanogen. The solution structure determined by NMR spectroscopy showed that it has a novel $\alpha+\beta$-fold with a highly acidic ligand binding pocket. Since MTH1880 maintains its ultra-stable structural characteristics at both high temperature and pressure, it has been considered as an excellent model for studying protein folding. To initiate the structural and folding study of MTH1880 in proving its unusual stability, we performed the site directed mutagenesis and biochemical analysis of MTH1880 mutants. Data from circular dichroism and NMR spectroscopy suggest that the point mutations perturbed the structural stability of protein even though the secondary structure is retained. This study will provide the useful information in understanding the role of participating residues during folding-unfolding process and our result will be used in designing further folding experiments for hyper-thermopile proteins like MTH1880.

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