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Published November 30, 2022 | Version v1
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Time-resolved β-lactam cleavage by L1 metallo-β-lactamase

Creators

  • 1. University of Chicago
  • 2. Argonne National Laboratory
  • 3. Worcester Polytechnic Institute

Description

Serial x-ray crystallography can uncover binding events, and subsequent chemical conversions occurring during enzymatic reaction. Here, we reveal the structure, binding and cleavage of moxalactam antibiotic bound to L1 metallo-β-lactamase (MBL) from Stenotrophomonas maltophilia. Using time-resolved serial synchrotron crystallography, we show the time course of β-lactam hydrolysis and determine ten snapshots (20, 40, 60, 80, 100, 150, 300, 500, 2000 and 4000 ms) at 2.20 Å resolution. The reaction is initiated by laser pulse releasing Zn2+ ions from a UV-labile photocage. Two metal ions bind to the active site, followed by binding of moxalactam and the intact β-lactam ring is observed for 100 ms after photolysis. Cleavage of β-lactam is detected at 150 ms and the ligand is significantly displaced. The reaction product adjusts its conformation reaching steady state at 2000 ms corresponding to the relaxed state of the enzyme. Only small changes are observed in the positions of Zn2+ ions and the active site residues. Mechanistic details captured here can be generalized to other MBLs.

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Additional details

Identifiers

DOI
10.1038/s41467-022-35029-3
Other
oai:uchicago.tind.io:5171

Funding

National Institute of Allergy and Infectious Diseases
HHSN272201700060C

UChicago Information

Division(s)
Biological Sciences Division
Department(s)
Biochemistry and Molecular Biology
Center(s) or Institute(s)
Center for Advanced Radiation Sources