Abstract
Methylotrophic yeast has previously been shown to be an excellent system for the cost-effective production of perdeuterated biomass and for the heterologous expression of membrane receptors. A protocol for the expression of 85% deuterated, functional human μ-opiate receptor was established. For partially deuterated biomass, deuteration level and distribution were determined for fatty acids, amino acids and carbohydrates. It was shown that prior to biosynthesis of lipids and amino acids (and of carbohydrates, to a lower extent), exchange occurs between water and methanol hydrogen atoms, so that 80%–90% randomly deuterated biomass and over-expressed proteins may be obtained using only deuterated water.
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Anglister, J., Scherf, T., Zilber, B. and Levy, R. (1995) Biopolymers, 37, 383–389.
Areas, J.A., Grobner, G., Glaubitz, C. and Watts, A. (1998) Biochemistry, 37, 5582–5588.
Augé, S., Marsan, M.P., Czaplicki, J., Demange, P., Muller, I., Tropis, M. and Milon, A. (1995) J. Chim. Phys., 92, 1715–1720.
Auger, M. (1997) Biophys. Chem., 68, 233–241.
Batey, R.T., Cloutier, N., Mao, H. and Williamson, J.R. (1996) Nucleic Acids Res., 24, 4836–4837.
Batey, R.T., Inada, M., Kujawinski, E., Puglisi, J.D. and Williamson, J.R. (1992) Nucleic Acids Res., 20, 4515–4523.
Befort, K., Tabbara, L., Bausch, S., Chavkin, C., Evans, C. and Kieffer, B. (1996) Mol. Pharmacol., 49, 216–223.
Bersch, B., Koehl, P., Nakatani, Y., Ourisson, G. and Milon, A. (1993) J. Biomol. NMR, 3, 443–461.
Blake, A.D., Bot, G., Tallent, M., Law, S.F., Li, S., Freeman, J.C. and Reisine, T. (1997) Receptors Channels, 5, 231–235.
Constantine, K.L., Mueller, L., Goldfarb, V., Wittekind, M., Metzler, W.J., Yanchunas Jr., J., Robertson, J.G., Malley, M.F., Friedrichs, M.S. and Farmer II, B.T. (1997) J. Mol. Biol., 267, 1223–1246.
Curley Jr., R.W., Panigot, M.J., Hansen, A.P. and Fesik, S.W. (1994) J. Biomol. NMR, 4, 335–340.
Czaplicki, J. and Milon, A. (1998) J. Chim. Phys., 95, 196–207.
Dahlquist, F.W., Muchmore, D.C., Davis, J.H. and Bloom, M. (1977) Proc. Natl. Acad. Sci. USA, 74, 5435–5439.
Davis, J.H. (1983) Biochim. Biophys. Acta, 737, 117–171.
DeJongh, D.C., Radford, T., Hribar, J.D., Hanessian, S., Bieber, M., Dawson, G. and Sweeley, C.C. (1969) J. Am. Chem. Soc., 91, 1728–1740.
Demange, P., Abdallah, M.A. and Frank, H. (1988) J. Chromatogr., 438, 291–297.
Dominguez, H., Rollin, C., Guyonvarch, A., Guerquin-Kern, J.-L., Cocaign-Bousquet, M. and Lindley, N.D. (1998) Eur. J. Biochem., 254, 96–102.
Farmer II, B.T., Constantine, K.L., Goldfarb, V., Friedrichs, M.S., Wittekind, M., Yanchunas Jr., J., Robertson, J.G. and Mueller, L. (1996) Nat. Struct. Biol., 3, 995–997.
Fejzo, J., Lepre, C.A., Peng, J.W., Su, M.S., Thomson, J.A. and Moore, J.M. (1996) Protein Sci., 5, 1917–1921.
Gallet, P.F., Vaujour, H., Petit, J.M., Maftah, A., Oulmouden, A., Oriol, R., Le Narvor, C., Guilloton, M. and Julien, R. (1998) Glycobiology, 8, 919–925.
Gardner, K.H., Rosen, M.K. and Kay, L.E. (1997) Biochemistry, 36, 1389–1401.
Gardner, K.H. and Kay, L.E. (1998) Annu. Rev. Biophys. Biomol. Struct., 27, 357–406.
Garrett, D.S., Seok, Y.J., Liao, D.I., Peterkofsky, A., Gronenborn, A.M. and Clore, G.M. (1997) Biochemistry, 36, 2517–2530.
Griffin, R.G. (1998) Nat. Struct. Biol., 5, 508–512.
Grobner, G., Choi, G., Burnett, I.J., Glaubitz, C., Verdegem, P.J., Lugtenburg, J. and Watts, A. (1998) FEBS Lett., 422, 201–204.
Haon, S., Augé, S., Tropis, M., Milon, A. and Lindley, N. (1993) J. Label. Comp. Radiopharm., 33, 1053–1063.
Hollenberg, C.P. and Gellissen, G. (1997) Curr. Opin. Biotechnol., 8, 554–560.
Huang, X., Yu, P., LeProust, E. and Gao, X. (1997) Nucleic Acids Res., 25, 4758–4763.
Kaiser, F.E., Gehrke, C.W., Zumwalt, R.W. and Kuo, K.C. (1974) J. Chromatogr., 94, 113–133.
Kay, L.E. and Gardner, K.H. (1997) Curr. Opin. Struct. Biol., 7, 722–731.
Kigawa, T., Muto, Y. and Yokoyama, S. (1995) J. Biomol. NMR, 6, 129–134.
Kim, I., Watanabe, S., Muto, Y., Hosono, K., Takai, K., Takaku, H., Kawai, G., Watanabe, K. and Yokoyama, S. (1995) Nucleic Acids Symp. Ser., 123–124.
Kushlan, D.M. and LeMaster, D.M. (1993) J. Biomol. NMR, 3, 701–708.
Laroche, Y., Storme, V., De Meutter, J., Messens, J. and Lauwereys, M. (1994) Biotechnology (NY), 12, 1119–1124.
Lee, B.J., Aiba, H. and Kyogoku, Y. (1991) Biochemistry, 30, 9047–9054.
LeMaster, D.M. (1987) FEBS Lett., 223, 191–196.
LeMaster, D.M. (1989) Methods Enzymol., 177, 23–43.
LeMaster, D.M. (1990a) Q. Rev. Biophys., 23, 133–174.
LeMaster, D.M. (1990b) Annu. Rev. Biophys. Biophys. Chem., 19, 243–266.
Markley, J.L., Putter, I. and Jardetzky, O. (1968) Science, 161, 1249–1251.
Marsan, M.P., Muller, I. and Milon, A. (1996a) Chem. Phys. Lipids, 84, 117–121.
Marsan, M.P., Warnock, W., Muller, I., Nakatani, Y., Ourisson, G. and Milon, A. (1996b) J. Org. Chem., 61, 4252–4257.
Massou, S., Augé, S., Tropis, M., Lindley, N.D. and Milon, A. (1998) J. Chim. Phys., 95, 406–411.
McCloskey, J.A. (1990) Methods Enzymol., 193, 329–338.
Metzler, W.J., Witteking, M., Goldfarb, V., Mueller, L. and Farmer II, B.T. (1996) J. Am. Chem. Soc., 118, 6800–6801.
Milon, A., Miyazawa, T. and Higashijima, T. (1990) Biochemistry, 29, 65–75.
Neri, D., Szyperski, T., Otting, G., Senn, H. and Wüthrich, K. (1989) Biochemistry, 28, 7510–7516.
Oda, Y., Nakamura, H., Yamazaki, T., Nagayama, K., Yoshida, M., Kanaya, S. and Ikehara, M. (1992) J. Biomol. NMR, 2, 137–147.
Ottleben, H., Haasemann, M., Ramachandran, R., Muller-Esterl, W. and Brown, L.R. (1997) Receptors Channels, 5, 237–241.
Pachter, R., Arrowsmith, C.H. and Jardetzky, O. (1992) J. Biomol. NMR, 2, 183–194.
Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366–12371.
Phongdara, A., Merckelbach, A., Keup, P., Gellissen, G. and Hollenberg, C.P. (1998) Appl. Microbiol. Biotechnol., 50, 77–84.
Prosser, R.S., Daleman, S.I. and Davis, J.H. (1994) Biophys. J., 66, 1415–1428.
Putter, I., Markley, J.L. and Jardetzky, O. (1970) Proc. Natl. Acad. Sci. USA, 65, 395–401.
Reat, V., Patzelt, H., Ferrand, M., Pfister, C., Oesterhelt, D. and Zaccai, G. (1998) Proc. Natl. Acad. Sci. USA, 95, 4970–4975.
Reisman, J.M., Hsu, V.L., Jariel-Encontre, I., Lecou, C., Sayre, M.H., Kearns, D.R. and Parello, J. (1993) Eur. J. Biochem., 213, 865–873.
Rosen, M.K., Gardner, K.H., Willis, R.C., Parris, W.E., Pawson, T. and Kay, L.E. (1996) J. Mol. Biol., 263, 627–636.
Shu, F., Ramakrishnan, V. and Schoenborn, B.P. (1996) Basic Life Sci., 64, 309–323.
Smith, B.O., Ito, I., Raine, A., Teichmann, S., Ben-Tovim, L., Nietlispach, D., Broadhurst, R.W., Terada, T., Kelly, M. and Oschkinat, K. (1996) J. Biomol. NMR, 8, 360–368.
Smith, D.E., Su, J.Y. and Jucker, F.M. (1997) J. Biomol. NMR, 10, 245–253.
Sreekrishna, K., Brankamp, R.G., Kropp, K.E., Blankenship, D.T., Tsay, J.T., Smith, P.L., Wierschke, J.D., Subramaniam, A. and Birkenberger, L.A. (1997) Gene, 190, 55–62.
Sudbery, P.E. (1996) Curr. Opin. Biotechnol., 7, 517–524.
Szyperski, T., Neri, D., Leiting, B., Otting, G. and Wüthrich, K. (1992) J. Biomol. NMR, 2, 323–334.
Talmont, F., Sidobre, S., Demange, P., Milon, A. and Emorine, L.J. (1996) FEBS Lett., 394, 268–272.
Tropis, M., Bardou, F., Bersch, B., Daffe, M. and Milon, A. (1996) Biochim. Biophys. Acta, 1284, 196–202.
Tschopp, J.F. and Cregg, J.M. (1991) Biotechnology, 18, 305–322.
Vanatalu, K., Paalme, T., Vilu, R., Burkhardt, N., Junemann, R., May, R., Ruhl, M., Wadzack, J. and Nierhaus, K.H. (1993) Eur. J. Biochem., 216, 315–321.
Venters, R.A., Huang, C.C., Farmer II, B.T., Trolard, R., Spicer, L.D. and Fierke, C.A. (1995) J. Biomol. NMR, 5, 339–344.
Waites, M.J., Lindley, N.D. and Quayle, J.R. (1981) J. Gen. Microbiol., 122, 193–199.
Wishart, D.S., Sykes, B.D. and Richards, F.M. (1993) Biochim. Biophys. Acta, 1164, 36–46.
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Massou, S., Puech, V., Talmont, F. et al. Heterologous expression of a deuterated membrane-integrated receptor and partial deuteration in methylotrophic yeasts. J Biomol NMR 14, 231–239 (1999). https://doi.org/10.1023/A:1008371118772
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DOI: https://doi.org/10.1023/A:1008371118772