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The relative catalytic efficiency of β-lactamase catalysed acyl and phosphyl transfer

Slater, Martin J., Laws, Andrew P. and Page, Michael I. (2001) The relative catalytic efficiency of β-lactamase catalysed acyl and phosphyl transfer. Bioorganic Chemistry, 29. pp. 77-95. ISSN 0045-2068

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Phosphonamidates which bear a simple resemblance to penicillin type structures have been synthesised as potential inhibitors of β-lactamases: -ethyl N-(benzyloxycarbonyl) amidomethyl phosphonyl amides, PhCH2OCONHCH2P(O)(OEt)NR2, the amines HNR2 being -proline, -proline, -thiazolidine, and o-anthranilic acid. The proline derivatives completely and irreversibly inactivated the class C β-lactamase from Enterobacter cloacae P99, in a time-dependent manner, indicative of covalent inhibition. The inactivation was found to be exclusive to the class C enzyme and no significant inhibition was observed with any other class of β-lactamase. The anthranilic acid derivative exhibited no appreciable inactivation of the β-lactamases. The phosphonyl proline and phosphonyl thioproline derivatives were separated into their diastereoisomers and their individual second order rate constants for inhibition were found to be 7.72 ± 0.37 and 8.3 × 10−2 ± 0.004 M−1 s−1 for the -proline derivatives, at pH 7.0. The products of the inhibition reaction of each individual diastereoisomer, analyzed by electrospray mass spectroscopy, indicate that the more reactive diastereoisomers phosphonylate the enzyme by P-N bond fission with the elimination of proline. Conversely, gas chromatographic detection of ethanol release by the less reactive proline diastereoisomer suggests phosphonylation occurs by P-O bond fission. The enzyme enhances the rate of phosphonylation with P-N fission by at least 106 compared with that effected by hydroxide-ion. The pH dependence of the rate of inhibition of the β-lactamase by the more reactive diasteroisomer is consistent with the reaction of the diprotonated form of the enzyme, EH2, with the inhibitor, I (or its kinetic equivalents EH with IH). This pH dependence and the rate enhancement indicate that the enzyme appears to use the same catalytic apparatus for phosphonylation as that used for hydrolysis of β-lactams. The stereochemical consequences of nucleophilic displacement at the phosphonyl centre are discussed

Item Type: Article
Subjects: Q Science > Q Science (General)
Q Science > QD Chemistry
Schools: School of Applied Sciences
School of Applied Sciences > Biomolecular Sciences Research Centre
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References: 1. Page, M. I., andWilliams, A. (1997) in Organic and Bioorganic Mechanisms, Longman, pp. 134–160. 2. Burgi, H. B., and Dunitz, J. D. (1983) Acc. Chem. Res. 16, 153. 3. Thatcher, G. R. J., and Kluger, R. (1989) Adv. Phys. Org. Chem. 25, 99. 4. Menger, F. M., (1985) Acc. Chem. Res. 18, 128; Menger, F. M., (1993) Acc. Chem. Res. 26, 206; Daffron, A., and Koshland, D. E. (1971) Proc. Natl. Acad. Sci. USA, 68, 2463; Kirby, A. J., (1997) Acc. Chem. Res. 30, 290; Kirby, A. J., (1996) Angew. Chem. Int. Ed. Engl. 35, 707. 5. Page, M. I., Laws, A. P., Slater, M. J., Stone, J. R., (1995) Pure Appl. Chem. 67, 711; Laws, A. P., Page, M. I., and Slater, M. J. (1993) Bioorg Med. Chem. Lett. 3, 2317. 6. Jacobsen, N. E., and Bartlett, P. A. (1981) J. Am. Chem. Soc. 103, 654. 7. Buckwell, S. C., Page, M. I., and Longridge, J. (1988) J. Chem. Soc. Perkin Trans 2, 1809. 8. Joris, B., De Meester, F., Galleni, M., Reckinger, G., Coyette, J., Fre`re, J.-M., and Van Beeumen, J. (1985) Biochem. J. 228, 241. 9. Waley, S. G. (1992) in The Chemistry of b-Lactams (Page, M. I., Ed.), Ch. 6, Chapman and Hall, London. 10. Oefner, C., D’Arcy, A., Daly, J. J., Gubernator, K., Charnas, R. L., Heinze, I., Hubschwerfen, C., and Winkler, F. K. (1990) Nature, 343, 284. 11. Lobkovsky, E., Moews, P. C., Lin, H., Zhao, H., Fre`re, J.-M., and Knox, J. R. (1993) Proc. Natl. Acad. Sci., USA, 90, 11257; Jelsch, C., Mourey, L., Masson, J.-M., and Samama, J.-P.(1993) Proteins: Struct. Funct. Genet. 16, 364; Moews, P. C., Knox, J. R., Dideberg, O., Charlier, P., and Fre`re, J.- M. (1990) Proteins: Struct., Funct., Genet. 7, 156; Herzberg, O., and Moult, J.-M. (1987) Science 236, 694; Herzberg, O. (1991) J. Mol. Biol. 217, 701; Dideberg, O., Charlier, P.,We´ry, J.-P., Dehottay, P., Dusart, J., Erpicum, T., Fre`re J.-M., and Ghuysen, J.-M. (1987) Biochem J. 245, 911; Jelsch, C., Mourey, L., Masson, J.-M., and Samama, J.-P. (1993) Proteins: Struct., Funct., Genet. 16, 364.12. Strynadka, N. C. J., Adachi, H., Jensen, S. E., Johns, K., Sielecki, A., Betzel, C., Sutoh, K., and James, M. N. G. (1992) Nature 359, 700; Chen, C. C. H., Rahil, J., Pratt, R. F., and Herzberg, O. (1993) J. Miol. Biol. 234, 165. 13. Ghuysen, J.-M. (1991) Annu. Rev. Microbiol. 45, 37; Matagne, A., and Fre`re, J.-M. (1995) Biochim. Biophys. Acta. 1246, 109; Lamotte-Brasseur, J., Knox, J., Kelly, J. A., Charlier, P., Fonze´, E., Dideberg, O., and Fre`re, J.-M. (1994) Biotechnol. Gen. Eng. Revs. 12, 189. 14. Laws, A. P., Page, M. I., and Slater, M. J. (1993) Bioorg. Med. Chem. Lett. 2317. 15. Fink, A. F. (1993) Chemtracts - Biochem. Mol. Biol. 3, 395. 16. Buckwell, S. C., and Page, J. I. (1987) Adv. Biosci. 65, 24. 17. Page, M. I., Vilanova, B., and Layland, N. J. (199) J. Am. Chem. Soc. 117, 12092. 18. Lobkovsky, E., Billings, E. M., Moews, P. C., Rahill, J., Pratt, R. F., and Knox, J. R. (1994) Biochemistry 33, 6762. 19. Dubus, A., Normark, S., Kania, M., and Page, M. G. P. (1994) Biochemistry 33, 8577; Dubus, A., Ledent, P., Lamotte-Brasseur, J., and Frere, J.-M. (1996) Proteins : Struct., Funct. and Gen. 25, 473. 20. Page, M. I. (1990) in Comp. Med. Chem., (Sammes, P. G., Ed.), Vol. 2, pp. 45–60, Pergamon, Oxford; Page, M. I. (1984) in The Chemistry of Enzyme Action (Page, M. I., Ed.), pp. 1–54, Elsevier, Amsterdam. 21. Laws, A. P., and Page, M. I. (1989) J. Chem. Soc. Perkin Trans. 2, 1577; Laws, A. P., Layland, N. J., Proctor, D. G., and Page, M. I. (1993) J. Chem. Soc. Perkin Trans. 2, 17. 22. Page, M. I., andWilliams, A. (1977) in Organic and Bioorganic Mechanisms, pp. 152–157, Longmans. 23. Kovach, I. M. (1988) J. Enzyme Inhibition 2, 199; Kovach, I. M., and Enyedy, E. J. (1998) J. Amer. Chem. Soc. 120, 258. 24. see however: Mock, W. L., and Chua, D. C. Y. (1995) J. Chem. Soc. Perkin Trans 2., 2069. 25. Murphy, B. P., and Pratt, R. F. (1988) Biochem. J. 256, 669.
Depositing User: Sara Taylor
Date Deposited: 14 Feb 2008 15:57
Last Modified: 28 Aug 2021 10:38


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