Abstract
The substrate specificity of α-lytic protease has been changed dramatically, with a concomitant increase in activity, by replacing an active-site Met with Ala. The substrate specificity of both this mutant and another similar mutant are extraordinarily broad. X-ray crystallographic analysis shows that structural plasticity, a combination of alternate side-chain conformations and binding-site flexibility, allows both large and small substrates to be well accommodated.
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References
Fersht, A. R. Enzyme Structure and Mechanism (Freeman, San Francisco, 1985).
Wells, J. A. Cunningham, B. C., Graycar, T. P. & Estell, D. A. Proc. natn. Acad. Sci. U.S.A. 84, 5167 (1987).
Estell, D. A. et al. Science 233, 659 (1986).
Wilks, H. M. et al. Science 242, 1541–1544 (1988).
Craik, C. S. et al. Science 228, 291–297 (1985).
Cronin, C. N. & Kirsch, J. F. Biochemistry 27, 4572 (1988).
Fujinaga, M., Delbaere, L. T. J., Brayer, G. D. & James, M. N. G. J. molec. Biol. 183, 479 (1985).
Brayer, G. D., Delbaere, L. T. J. & James, M. N. G. J. molec. Biol. 131, 743 (1979).
Hunkapiller, M. W., Forgac, M. D. & Richards, J. H. Biochemistry 15, 5581 (1976).
Bachovchin, W. W., Kaiser, R., Richards, J. H. & Roberts, J. D. Proc. natn. Acad. Sci. U.S.A. 78, 7323 (1981).
Robillard, G. & Schulman, R. G. J. molec. Biol. 86, 541 (1974).
Bone, R., Shenvi, A. B., Kettner, C. A. & Agard, D. A. Biochemistry 27, 7609 (1987).
Schecter, I. & Berger, A. Biochem. Biophys. Res. Commun. 27, 157 (1967).
Bone, R., Frank, D., Kettner, C. A. & Agard, D. A. Biochemistry (in the press).
Bauer, C.-A., Brayer, G. D., Sielecki, A. R. & James, M. N. G. Eur. J. Biochem. 120, 289–294 (1981).
Kettner, C. A., Bone, R., Agard, D. A. & Bachovchin, W. W. Biochemistry 27, 7682–7688 (1988).
Dorovska, V. M., Variolomegev, S. D., Kazanskaya, N. F., Klyosov, A. A. & Martinek, K. FEBS Lett. 23, 122 (1972).
Harper, W. J., Cook, R. R., Roberts, C. J., McLaughlin, B. J. & Powers, J. C. Biochemistry 23, 2995 (1984).
Nakajima, K., Powers, J. C., Ashe, B. M. & Zimmerman, M. J. Biol. Chem. 254, 4027 (1979).
Connolly, M. L. J. Appl. Crystallogr. 16, 548 (1983).
Connolly, M. L. Science 221, 709 (1983).
Silen, J. L., Frank, D., Fujishige, A., Bone, R. & Agard, D. A. J. Bacteriol. 171, 1320 (1989).
Wells, J. A., Vasser, M. & Powers, D. B. Gene 34, 315 (1985).
Whitaker, D. R. Meth. Enzymol. 19, 599 (1970).
Hunkapiller, M. W., Smallcombe, S. H., Whitaker, D. R. & Richards, J. H. Biochemistry 12, 4732 (1973).
Williams, J. W. & Morrison, J. F. Meth. Enzymol. 63, 437–467 (1979).
Hendrickson, W. A. & Konnert, J. in Biomolecular Structure, Function, Conformation, and Evolution (ed. Srinivasam, R.), Vol. 1, 43–47 (Pergamon, Oxford, 1981).
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Bone, R., Silen, J. & Agard, D. Structural plasticity broadens the specificity of an engineered protease. Nature 339, 191–195 (1989). https://doi.org/10.1038/339191a0
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DOI: https://doi.org/10.1038/339191a0
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