Abstract
Phytases catalyze the hydrolysis of phytate and are able to improve the nutritional quality of phytate-rich diets. Escherichia coli phytase, a member of the histidine acid phosphatase family has the highest specific activity of all phytases characterized. The crystal structure of E. coli phytase has been determined by a two-wavelength anomalous diffraction method using the exceptionally strong anomalous scattering of tungsten. Despite a lack of sequence similarity, the structure closely resembles the overall fold of other histidine acid phosphatases. The structure of E. coli phytase in complex with phytate, the preferred substrate, reveals the binding mode and substrate recognition. The binding is also accompanied by conformational changes which suggest that substrate binding enhances catalysis by increasing the acidity of the general acid.
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References
Reddy, N.R., Sathe, S.K. & Salunkhe, D.K. Adv. Food Res. 28, 1– 92 (1982).
Graf, E. In Phytic Acid Chemistry and Application (ed. Graf, E.) 1– 21 (Pilatus Press, Minneapolis; 1986).
Wodzinski, R.J. & Ullah, H.J. Adv. Appl. Micro. 42, 263–302 ( 1996).
Greiner, R., Konietzny, U. & Jany, K.-D. Arch. Biochem. Biophys. 303, 107–113 (1993).
Wyss, M. et al. Appl. Env. Microbiol. 65, 367– 373 (1999).
Chi, H. et al. Genomics 56, 324–336 (1999).
Vincent, J.B., Crowder, M.W. & Averill, B.A. Trends Biochem. Sci. 17, 105 –110 (1992).
Van Etten, R.L. Ann. N.Y. Acad. Sci. 390, 27–51 (1992).
Ostanin, K. et al. J. Biol. Chem. 267, 22830– 22836 (1992).
Ostanin, K. & Van Etten, R.L. J. Biol. Chem. 268 , 20778–20784 (1993).
Lindqvist, Y., Schneider, G. & Vihko, P. Eur. J. Biochem. 221, 139– 142 (1994).
Schneider, G., Lindqvist, V. & Vihko, P. EMBO J. 12, 2609– 2615 (1993).
LaCount, M.W., Handy, G. & Lebioda, L. J. Biol. Chem. 273, 30406– 30409 (1998).
Kostrewa, D. et al. Nature Struct. Biol. 4, 185– 190 (1997).
Kostrewa, D., Wyss, M., D'Arcy, A., van Loon, A.P.G.M. J. Mol. Biol. 288, 965–974 (1999).
Cohen, G.H. J. Appl. Crystallogr. 30, 1160–1161 (1997).
Jia, Z., Golovan, S., Ye, Q. & Forsberg, C.W. Acta Crystallogr. D54, 647–649 ( 1998).
Otwinowski, Z. In Proceedings of the CCP4 Study Weekend: Data Collection and Processing (ed Sawyer L., Issacs N.& Bailey S.) 56– 62 (Daresbury Laboratory, Warrington; 1993).
Minor, W. XdisplayF Program. Purdue University, West Lafayette, USA (1993 ).
Collaborative Computational Project, Number 4. Acta Crystallogr. D50, 760–763 (1994).
Egloff, M.-P., Cohen, P. T. W., Reinemer, P. & Barford, D. J. Mol. Biol. 254, 942–959 (1995).
de La Fortelle, E. & Bricogne, G. In Methods in Enzymology, Macromolecular Crystallography (eds. Sweet, R.M. & Carter, Jr. C.W.) 276, 472–494 (Academic Press, New York; 1997).
Abrahams J.P. & Leslie A.G.W. Acta Crystallogr. D52, 30–42 (1996).
McRee, D.E. J. Mol. Graphics 10, 44–47 (1992).
Brünger, A.T. et al. Acta Crystallogr. D54, 905– 921 (1998).
Navaza, J. Acta Crystallogr. A50, 157–163 (1994).
Laskowski, R.A., MacArthur, M.W., Moss, D.S. & Thornton, J.M. J. App. Crystallogr. 26, 283–291 (1993).
Engelen, A.J., Vanderheeft, F.C., Randsdorp, P.H.G., & Smit, E.L.C. J. AOAC. Int. 77: 760–764. (1994).
Provenzano, M.D., Fujimoto, E.K., Goeke, N.M., Olson, B.J., & Klenk, D.C. Anal. Biochem. 150: 76–85. (1985).
Kraulis, P.J. J. Appl. Crystallogr. 24, 946–950 (1992).
Merrit, E.A. & Murphy, M.E.P. Acta Crystallogr. D50, 869–873 (1994).
Nicholls, A., Sharp, K. & Honig, B. Proteins 11, 281– 296 (1991).
Acknowledgements
The authors would like to thank Y.-C. Liou of P. Davies' group at Queen's University for equipment and technical assistance in early purification attempts and for his support throughout the project. We thank A. Tocilj for his encouragement and assistance at various stages of the project. We thank A. Iyo and M. Cotrill for constructing and help with purifying the inactive mutant, respectively. We also thank E. Leinala for her advice on crystallization. G. Thatcher provided helpful discussion. We are grateful to L. Flaks and the technical staff at the National Synchrotron Light Source at Brookhaven National Laboratory for their support at X8-C. D.L. was an Ontario Graduate Scholarship recipient. This work was supported by a NSERC grant to Z.J. and contract funding from Ontario Pork to C.W.F.
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Lim, D., Golovan, S., Forsberg, C. et al. Crystal structures of Escherichia coli phytase and its complex with phytate. Nat Struct Mol Biol 7, 108–113 (2000). https://doi.org/10.1038/72371
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DOI: https://doi.org/10.1038/72371
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