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Molecular structure of the acyl-enzyme intermediate in β-lactam hydrolysis at 1.7 Å resolution

Nature volume 359, pages 700–705 (1992)Cite this article

Abstract

The X-ray crystal structure of the molecular complex of penicillin G with a deacylation-defective mutant of the RTEM-1 β-lactamase from Escherichia coli shows how these antibiotics are recognized and destroyed. Penicillin G is covalently bound to Ser 70 Oγ as an acyl-enzyme intermediate. The deduced catalytic mechanism uses Ser 70 Oγ as the attacking nucleophile during acylation. Lys 73 Nζ acts as a general base in abstracting a proton from Ser 70 and transferring it to the thiazolidine ring nitrogen atom via Ser 130 Oγ. Deacylation is accomplished by nucleophilic attack on the penicilloyl carbonyl carbon by a water molecule assisted by the general base, Glu 166.

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Authors and Affiliations

  1. MRC Group in Protein Structure and Function, Department of Biochemistry, University of Alberta, Edmonton, Alberta, T6G 2H7, Canada

    Natalie C. J. Strynadka, Kathy Johns, Anita Sielecki & Michael N. G. James

  2. Department of Pure and Applied Sciences, College of Arts and Sciences, University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo, 153, Japan

    Hiroyuki Adachi & Kazuo Sutoh

  3. Department of Microbiology, University of Alberta, Edmonton, Alberta, T6G 2E9, Canada

    Susan E. Jensen

  4. European Molecular Biology Laboratory, c/o DESY, Notkestrasse 85, Hamburg, Germany

    Christian Betzel

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Strynadka, N., Adachi, H., Jensen, S. et al. Molecular structure of the acyl-enzyme intermediate in β-lactam hydrolysis at 1.7 Å resolution. Nature 359, 700–705 (1992). https://doi.org/10.1038/359700a0

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