Abstract
Are the structure and function of a protein in the crystal the same as in solution? The question, put forward at the first publications of the three dimensional structure of proteins [1], is still dramatically valid. In fact, about ten thousand protein structures have been determined by X-ray crystallography, and more are exponentially coming out, but only 5% of these proteins have been tested whether their functional properties are maintained in the crystal. There is no a priori answer and each protein has to be individually investigated. A powerful technique for the determination of functional properties of proteins in the crystal is polarized absorption microspectrophotometry [2]. The chromophoric properties of cofactors, coenzymes, ligands, substrates and substrate analogs are exploited to monitor function-related spectroscopic properties and to determine kinetic and thermodynamic parameters. Two case-studies, crystals of T state hemoglobin and pyridoxal 5′-phosphate-dependent O-acetylserine sulfhydrylase, will allow to evidence some of the advantages of carrying out functional studies in the same physical state where the three dimensional structure is obtained.
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References
Rossi, G. L. and Bernhard, S. A., J. Mol. Biol. 49 (1970) 85
Mozzarelli, A. and Rossi, G. L., Annu. Rev. Biophys. Biomol. Struct. 25 (1996) 343
Monod, J., Wyman, J. and Changeux, J-P., J. Mol. Biol. 12 (1965) 88
Perutz, M. F., Nature 228 (1970) 726; Perutz, M. F., Wilkinson, A. J., Paoli, M. and Dodson, G., Annu. Rev. Biophys. Biomol. Struct. 27 (1998) 1
Koshland, D. E., Nemethy, G. and Filmer, D., Biochemistry 5 (1966) 365
Ackers, G., Doyle, M. L., Myers, D. and Daugherty, M. A., Science 255 (1992) 54
Mozzarelli, A., Rivetti, C., Rossi, G. L., Henry, E. R. and Eaton, W. A., Nature 351 (1991) 416; Rivetti, C., Mozzarelli, A., Rossi, G. L., Henry, E. R. and Eaton, W. A., Biochemistry 32 (1993) 2888; Eaton, W. A., Henry, E. R., Hofrichter, J. and Mozzarelli, A., Nature Struct. Biol. in press
Brzozowski, A., Derewenda, Z, Dodson, E., Dodson, G., Grabowski, M., Liddington, R., Skarzynski, T and Vallely, D, Nature 307 (1984) 725; Paoli, M., Liddington, R, Tame, J., Wilkinson, A. and Dodson, G., J. Mol. Biol. 256 (1996) 775
Shibayama, N. and Saigo, S. J. Mol. Biol. 251 (1995) 203; Bettati, S. and Mozzarelli, A., J. Biol. Chem. 272 (1997) 32050
Cook, P. F., Hara, S., Nalabolu, S. and Schnackerz, K. D., Biochemistry 31 (1992) 2298
Mozzarelli, A., Bettati, S., Pucci, A. M., Burkhard, P and Cook, P. F., J. Mol. Biol. 283 (1998) 135
Burkhard, P., Rao, G. S. J., Hohenester, E., Schnackerz, K. D., Cook, P. F. and Jansonius, J. N., J. Mol. Biol. 283 (1998) 121
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Mozzarelli, A., Bettati, S. (1999). Protein structure-function relationship studied by single crystal polarized absorption microspectrophotometry. In: Greve, J., Puppels, G.J., Otto, C. (eds) Spectroscopy of Biological Molecules: New Directions. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-4479-7_1
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DOI: https://doi.org/10.1007/978-94-011-4479-7_1
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