Abstract
A method is described for modifying G proteins with biotin. With transducin, better results were obtained with the amino group-specific derivative BXNHS (biotinyl-ε-aminocaproic acid N-hydroxysuccinimide ester) as compared to the -SH-group specific reagent MBB (maleimidobutyrylbiocytin). Modification occurred under conditions preserving functional activity: Interaction of the biotinylated transducin with rod outer segment membranes was shown by its light-dependent association and by a GTPγS-binding assay. G0 from bovine brain was also biotinylated under conditions preserving its activity. Biotinyl-α0 was shown to bind to a streptavidin Sepharose matrix. Biotinyl-G proteins, therefore, are proposed as tools for extracting proteins (receptors and effector systems), which interact under specific conditions with G proteins.
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References
Baehr, W., Morita, E. A., Swanson, R. J., and Applebury, M. L. (1982).J. Biol. Chem. 257, 6452–6460.
Birnbaumer, L., Codina, J., Mattera, R., Yatani, A., Graf, R., Olate, J., Sanford, J., and Brown, A. M. (1988).Cold Spring Harbor Symp. Quant. Biol. 80, 229–239.
Birnbaumer, L., Yatani, A., Codina, J., van Dongen, A., Graf, A., Mattera, R., Sanford, J., and Brown, A. M. (1989). In40 Colloquium Mosbach (Gehring, U., Helmreich, E., and Schultz, G., eds.), Springer-Verlag, Berlin, pp. 147–177.
Cerione, R. A., Codina, J., Benovic, J. L., Lefkowitz, R. J., Birnbaumer, L., and Caron, M. G. (1984).Biochem. 23, 4519–4525.
Cerione, R. A., Staniszweski, C., Benovic, J. L., Lefkowitz, R. J., Gierschik, P., Somers, R., Spiegel, A., Codina, J., and Birnbaumer, L. (1985).J. Biol. Chem. 260, 1493–1500.
Cerione, R. A., Regan, J. W., Nakata, H., Codina, J., Benovic, J. L., Gierschik, P., Somers, R. L., Spiegel, A. M., Birnbaumer, L., Lefkowitz, R. J., and Caron, M. G. (1986).J. Biol. Chem. 261, 3901.
Florio, V. A., and Sternweis, P. C. (1989).J. Biol. Chem. 264, 3909–3915.
Gilman, A. G. (1987).Ann. Rev. Biochem. 56, 615–649.
Kaziro, Y., Itoh, H., Kozasa, T., Toyama, R., Tsukamoto, T., Matsuoka, M., Nakafuku, M., Obara, T., Takagi, T., and Hernandez, R. (1988). InCold Spring Harbor Symposia on Quantitative Biology, Vol. 53, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp. 209–220.
Kohnken, R. E., and Hildebrandt, J. D. (1989).J. Biol. Chem. 264, 20,688–20,696.
Kühn, H., and Hargrave, P. A. (1981).Biochem. 20, 2410–2417.
Limbird, L. E. (1988).FASEB J. 2, 2686–2695.
Lochrie, M. A., and Simon, M. I. (1988).Biochem. 27, 4957–4965.
Neer, E. J., and Clapham, D. E. (1988).Nature 333, 129–134.
Northup, J. K., Smigel, M. D., and Gilman, A. G. (1982).J. Biol. Chem. 257, 11,416–11,423.
Papermaster, D. S., and Dreyer, W. J. (1974).Biochem. 13, 2438–2444.
Senogles, S. E., Spiegel, A. M., Padrell, E., Iyengar, R., and Caron, M. G. (1990).J. Biol. Chem. 265, 4507–4514.
Sunyer, J., Monastirsky, B., Codina, J., and Birnbauer, L. (1989).Mol. Endocrinol. 3, 1115–1124.
Ui, M. (1984).TIPS 5, 277–279.
Updyke, T. V., and Nicolson, G. L. (1984).J. Immunol. Methods 73, 83–95.
Van Dongen, A. M., Codina, J., Olate, J., Mattera, R., Joho, R., Birnbaumer, L., and Brown, A. M. (1988).Science 242, 1433–1437.
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Heller, C., Hucho, F. Biotinylation of transducin and G0 from bovine brain. J Protein Chem 10, 325–332 (1991). https://doi.org/10.1007/BF01025631
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DOI: https://doi.org/10.1007/BF01025631