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1 pp. Why are thermophilic proteins thermally stable Satoshi Akanuma, Akihiko Yamagishi Department of Molecular Biology, Tokyo University of Pharmacy and Life Sciences, Horinouchi, Hachioji, Tokyo, Japan N U GNU GNU

2 N U K GNU GNU HNU T SNU GNU GNU GNU Tm Tm Tm Tm GNU Tm Thermus thermophilus - IPMDH IPMDH Tm Tm GNU Tm Tm M Tm T Tm Tm Tm Tm N U X ki U X N X kobs t kobs ku kikf ki t ln kobs

3 kobs t kobs M kobs T t M t T kobs t kobs t kobs t Sulfolobus islandicus DNA ORF Thermotoga maritima LDH LDH Haney Methanococcus jannaschii Methanocaldococcus jannaschii Methanococcus Methanococcus GC Gln Asn Thr SerGln Asn Ser Thr Glu Arg Lys α α α α α α α Glu Met Ala Leu α Petukhov Facchiano α α α α GNU Thermococcus ko-

4 α α NH CO α α dakaraensis HII Dong Ile Leu Ala Cp Cp GNU β Pyrococcus horikoshii CutA CutA T. maritima DNA HU Tm Arg LysGluLys Bacillus stearothermophilus HU Tm GluLys Pyrococcus furiosus P. furiosus Lys Glu Greaves Warwicker Kumer Nussinov IPMDH

5 - IPMDH Tm T. thermophilus IPMDH A IPMDH Tm S. tokodaii IPMDH B IPMDH Sulfolobus tokodaii IPMDH S. tokodaii IPMDH T. thermophilus IPMDH α IPMDH Leu Val Glu Met IPMDH IPMDH Leu Val IPMDH P. horikoshii PhAcP PhAcP IPMDH Bacillus amyloliquefaciens α-tm B. licheniformis α-tm T. thermophilus IPMDH Cp

6 - IPMDH A T. thermophilus IPMDH α B T. thermophilus IPMDH IPMDH T. thermophilus IPMDH Leu Val IPMDH Glu Met IPMDH Závodszky T. thermophilus IPMDH H D H D IPMDH IPMDH T P. furiosus Clostridium pasteurianum Tm B. amyloliquefaciens α-b. licheniformis α-

7 Cys Cys Akanuma, S., Yamagishi, A., Tanaka, N., & Oshima, T. Protein Sci.,,. Kumar, S., Tsai, C.J., & Nussinov, R.Biochemistry,,. Sterner, R. & Liebl, W.Crit. Rev. Biochem. Mol. Biol.,,. Razvi, A. & Scholtz, J.M.Protein. Sci.,,. Nojima, H., Ikai, A., Oshima, T., & Noda, H.J. Mol. Biol.,,. Motono, C., Oshima, T., & Yamagishi, A.Protein. Eng.,,. Li, W.T., Grayling, R.A., Sandman, K., Edmondson, S., Shriver, J.W., & Reeve, J.N.Biochemistry,,. Luke, K.A., Higgins, C.L., & Wittung-Stafshede, P. FEBS. J.,,. Berezovsky, I.N. & Shakhnovich, E.I. Proc. Natl. Acad. Sci. USA,,. Zeeb, M., Lipps, G., Lilie, H., & Balbach, J.J. Mol. Biol.,,. Perl, D., Welker, C., Schindler, T., Schroder, K., Marahiel, M. A., Jaenicke, R., & Schmid, F.X.Nat. Struct. Biol.,,. Auerbach, G., Ostendorp, R., Prade, L., Korndorfer, I., Dams, T., Huber, R., & Jaenicke, R.Structure,,. Haney, P.J., Badger, J.H., Buldak, G.L., Reich, C.I., Woese, C. R., & Olsen, G.J.Proc. Natl. Acad. Sci. USA,,. Wright, H.T.Crit. Rev. Biochem. Mol. Biol.,,. Chou, P.Y. & Fasman, G.D.Adv. Enzymol. Relat. Areas. Mol. Biol.,,. Petukhov, M., Kil, Y., Kuramitsu, S., & Lanzov, V. Proteins,,. Facchiano, A.M., Colonna, G., & Ragone, R.Protein. Eng.,,. Kahsai, M.A., Martin, E., Edmondson, S.P., & Shriver, J.W. Biochemistry,,. Dong, H., Mukaiyama, A., Tadokoro, T., Koga, Y., Takano, K., & Kanaya, S.J. Mol. Biol.,,. Chakravarty, S. & Varadarajan, R.FEBS. Lett.,,. Thompson, M.J. & Eisenberg, D.J. Mol. Biol.,,. Cambillau, C. & Claverie, J.M.J. Biol. Chem.,,. Tanaka, T., Sawano, M., Ogasahara, K., Sakaguchi, Y., Bagautdinov, B., Katoh, E., Kuroishi, C., Shinkai, A., Yokoyama, S.,&Yutani,K.FEBS. Lett.,,. Christodoulou, E., Rypniewski, W.R., & Vorgias, C.R. Extremophiles,,. Arnott, M.A., Michael, R.A., Thompson, C.R., Hough, D.W., & Danson, M.J.J. Mol. Biol.,,. Strop, P. & Mayo, S.L.Biochemistry,,. Kumar, S., Ma, B., Tsai, C.J., & Nussinov, R.Proteins,,. Greaves, R.B. & Warwicker, J.BMC. Struct. Biol.,,. Kumar, S. & Nussinov, R.Chembiochem.,,. Goodsell, D.S. & Olson, A.J.Annu. Rev. Biophys. Biomol. Struct.,,. Maes, D., Zeelen, J.P., Thanki, N., Beaucamp, N., Alvarez, M., Thi, M.H., Backmann, J., Martial, J.A., Wyns, L., Jaenicke, R., & Wierenga, R.K.Proteins,,. Villeret, V., Clantin, B., Tricot, C., Legrain, C., Roovers, M., Stalon, V., Glansdorff, N., & Van Beeumen, J.Proc. Natl. Acad. Sci. USA,,. Akanuma, S., Yamagishi, A., Tanaka, N., & Oshima, T. Eur. J. Biochem.,,. Kirino, H., Aoki, M., Aoshima, M., Hayashi, Y., Ohba, M., Yamagishi, A., Wakagi, T., & Oshima, T.Eur. J. Biochem.,,. Cheung, Y.Y., Lam, S.Y., Chu, W.K., Allen, M.D., Bycroft, M., & Wong, K.B.Biochemistry,,. Fitter, J. & Haber-Pohlmeier, S. Biochemistry,,. Robic, S., Guzman-Casado, M., Sanchez-Ruiz, J.M., & Marqusee, S.Proc. Natl. Acad. Sci. USA,,. Zavodszky, P., Kardos, J., Svingor, & Petsko, G.A. Proc. Natl. Acad. Sci. USA,,. Wray, J.W., Baase, W.A., Lindstrom, J.D., Weaver, L.H., Poteete, A.R., & Matthews, B.W.J. Mol. Biol.,,.

8 LeMaster, D.M., Tang, J., Paredes, D.I., & Hernandez, G. Proteins,,. Matsumura, M., Becktel, W.J., Levitt, M., & Matthews, B.W. Proc. Natl. Acad. Sci. USA,,. Ivens, A., Mayans, O., Szadkowski, H., Jurgens, C., Wilmanns, M., & Kirschner, K.Eur. J. Biochem.,,. Watanabe, K., Ohkuri, T., Yokobori, S., & Yamagishi, A. J. Mol. Biol.,,. Iwabata, H., Watanabe, K., Ohkuri, T., Yokobori, S., & Yamagishi, A.FEMS Microbilo. Lett.,,. Shimizu, H., Yokobori, S., Ohkuri, T., Yokogawa, T., Nishikawa, T., & Yamagishi, A.J. Mol. Biol.,,.

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