Summary
A novel protocol for isotopically labeling bacterially expressed proteins is presented. This method circumvents problems related to poor cell growth, commonly associated with the use of minimal labeled media, and problems with protein induction encountered, less commonly, when using enriched labeled media. The method involves initially growing the bacterial cells to high optical density in a commercially available enriched labeled medium. Following a suitable growth period, the cells are transferred to a different (minimal) labeled medium, appropriate for induction. The method is demonstrated using the protein melanoma growth stimulating activity (MGSA).
References
Bax, A. and Grzesiek, S. (1993) Acc. Chem. Res., 26, 131–138.
Bax, A., Ikura, M., Kay, L.E., Torchia, D.A. and Tschudin, R. (1990) J. Magn. Reson., 86, 304–318.
Clore, G.M. and Gronenborn, A.M. (1991a) Prog. NMR Spectrosc., 23, 43–92.
Clore, G.M. and Gronenborn, A.M. (1991b) Science, 252, 1390–1399.
Fairbrother, W.J., Reilly, D., Colby, T. and Horuk, R. (1993) FEBS Lett., 330, 302–306.
Horuk, R., Yansura, D.G., Reilly, D., Spencer, S., Bourell, J., Henzel, W., Rice, G. and Unemori, E. (1993) J. Biol. Chem., 268, 541–546.
Kikuchi, Y., Yoda, K., Yamasaki, M. and Tamura, G. (1981) Nucleic Acids Res., 9, 5671–5678.
McIntosh, L.P. and Dahlquist, F.W. (1990) Q. Rev. Biophys., 23, 1–38.
Norwood, T.J., Boyd, J., Heritage, J.E., Soffe, N. and Campbell, I.D. (1990) J. Magn. Reson., 87, 488–501.
Torriani, A. (1960) Biochim. Biophys. Acta, 38, 460–479.
Wagner, G. (1993) J. Biomol. NMR, 3, 375–385.
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Reilly, D., Fairbrother, W.J. A novel isotope labeling protocol for bacterially expressed proteins. J Biomol NMR 4, 459–462 (1994). https://doi.org/10.1007/BF00179352
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DOI: https://doi.org/10.1007/BF00179352