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Trapping and visualization of a covalent enzyme–phosphate intermediate

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Using a mutant version of E. coli alkaline phosphatase, we succeeded in trapping and determining the structure of the phospho-enzyme intermediate. The X-ray structure also revealed the catalytic water molecule, bound to one of the active site zinc ions, positioned ideally for the apical attack necessary for the hydrolysis of the intermediate.

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References

  1. Stoddard, B.L. Pharmocol Ther. 70, 215–256 (1996).

    Article  CAS  Google Scholar 

  2. Genick, U.K. et al. Science 275, 1471–1475 (1997).

    Article  CAS  Google Scholar 

  3. Bolduc, J.M. et al. Science 268, 1312–1318 (1995).

    Article  CAS  Google Scholar 

  4. Verschueren, K.H., Seljee, F., Rozeboom, H.J., Kalk, K.H. & Dijkstra, B.W. Nature 363, 693–698 (1993).

    Article  CAS  Google Scholar 

  5. Kallarakal, A.T. et al. Biochemistry 34, 2788–2797 (1995).

    Article  CAS  Google Scholar 

  6. Scott, W.G., Murray, J.B., Arnold, J.R.P., Stoddard, B.L. & Klug, A. Science 274, 2065–2069 (1996).

    Article  CAS  Google Scholar 

  7. Ding, X., Rasmussen, B.F., Petsko, G.A. & Ringe, D. Biochemistry 33, 9285–9293 (1994).

    Article  CAS  Google Scholar 

  8. Yennawar, N.H., Yennawar, H.P. & Farber, G.K. Biochemistry 33, 7326–7336 (1994).

    Article  CAS  Google Scholar 

  9. Schwartz, J.H. & Lipmann, F. Proc. Natl. Acad. Sci. USA 47, 1996–2005 (1961).

    Article  CAS  Google Scholar 

  10. Bloch, W. & Gorby, M.S. Biochemistry 19, 5008–5018 (1980).

    Article  CAS  Google Scholar 

  11. Gettins, P. & Coleman, J.E. J. Biol. Chem 258, 408–416 (1983).

    PubMed  CAS  Google Scholar 

  12. Hull, W.E., Halford, S.E., Gutfreund, H. & Sykes, B.D. Biochemistry 15, 1547–1561 (1976).

    Article  CAS  Google Scholar 

  13. Reid, T.W. & Wilson, I.B. Enzymes (3rd Ed.) 4, 373–415 (1971).

    Article  CAS  Google Scholar 

  14. Kim, E.E. & Wyckoff, H.W. J. Mol. Biol. 218, 449–464 (1991).

    Article  CAS  Google Scholar 

  15. Ko, S.H.D. & Kézdy, F.J. J. Amer. Chem. Soc. 89, 7139–7140 (1967).

    Article  CAS  Google Scholar 

  16. Bloch, W. & Schlesinger, M.J. J. Biol. Chem. 248, 5794–5805 (1973).

    PubMed  CAS  Google Scholar 

  17. Bale, J.R., Huang, C.Y. & Chock, P.B. J. Biol. Chem. 255, 8431–8436 (1980).

    PubMed  CAS  Google Scholar 

  18. Engström, L. Biochim. Biophys. Acta 56, 606–609 (1962).

    Article  Google Scholar 

  19. Milstein, C. Biochem. J. 92, 410–419 (1964).

    Article  CAS  Google Scholar 

  20. Pigretti, M.M. & Milstein, C. Biochem. J. 94, 106–113 (1965).

    Article  Google Scholar 

  21. Murphy, J.E., Tibbitts, T.T. & Kantrowitz, E.R. J. Mol. Biol. 253, 604–617 (1995).

    Article  CAS  Google Scholar 

  22. Denu, J.M., Lohne, D.L., Vijayalakshmi, J., Saper, M.A. & Dixon, J.E. Proc. Natl. Acad. Sci. USA 93, 2493–2498 (1996).

    Article  CAS  Google Scholar 

  23. Ma, L. & Kantrowitz, E.R. J. Biol. Chem. 269, 31614–31619 (1994).

    PubMed  CAS  Google Scholar 

  24. Xu, X. & Kantrowitz, E.R. Biochemistry 30, 7789–7796 (1991).

    Article  CAS  Google Scholar 

  25. Janeway, C.M.L., Xu, X., Murphy, J.E., Chaidaroglou, A. & Kantrowitz, E.R. Biochemistry 32, 1601–1609 (1993).

    Article  CAS  Google Scholar 

  26. Xu, X., Qin, X.-Q. & Kantrowitz, E.R. Biochemistry 33, 2279–2284 (1994).

    Article  CAS  Google Scholar 

  27. Brünger, A.T. X-PLOR, Version 3.1 (Yale University Press, New Haven, Connecticut; 1992).

  28. Engh, R.A. & Huber, R. Acta Crystallogr. A47, 392–400 (1991).

    Article  CAS  Google Scholar 

  29. Brünger, A.T. Nature 355, 472–475 (1992).

    Article  Google Scholar 

  30. Evans, S.V. J. Mol. Graph. 11, 134–138 (1993).

    Article  CAS  Google Scholar 

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Author notes

  1. Jennifer E. Murphy

    Present address: Jennifer M. Schneiders, Yale University School of Medicine, Department of Molecular Biophysics and Biochemistry, P.O. Box 208024, 333 Cedar Street, New Haven, Connecticut, 06520-8024, USA

Authors and Affiliations

  1. Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, Massachusetts, 02167, USA

    Boguslaw Stec, Lan Ma & Evan R. Kantrowitz

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  1. Jennifer E. Murphy
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  2. Boguslaw Stec
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  3. Lan Ma
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Murphy, J., Stec, B., Ma, L. et al. Trapping and visualization of a covalent enzyme–phosphate intermediate. Nat Struct Mol Biol 4, 618–622 (1997). https://doi.org/10.1038/nsb0897-618

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