Time-resolved β-lactam cleavage by L1 metallo-β-lactamase

Wilamowski, M.; Sherrell, D. A.; Kim, Y.; Lavens, A.; Henning, R. W.; Lazarski, K.; Shigemoto, A.; Endres, M.; Maltseva, N.; Babnigg, G.; Burdette, S. C.; Srajer, V.; Joachimiak, A.

doi:10.1038/s41467-022-35029-3

Title: Time-resolved β-lactam cleavage by L1 metallo-β-lactamase

Journal Article · Wed Nov 30 00:00:00 EST 2022 · Nature Communications

DOI:https://doi.org/10.1038/s41467-022-35029-3· OSTI ID:1901479

; Sherrell, D. A.;

;

; Henning, R. W.; Lazarski, K.; Shigemoto, A.; Endres, M.;

;

; Srajer, V.;

Abstract 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 Zn ²⁺ 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 Zn ²⁺ ions and the active site residues. Mechanistic details captured here can be generalized to other MBLs.

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Sponsoring Organization:: USDOE

OSTI ID:: 1901479

Journal Information:: Nature Communications, Journal Name: Nature Communications Vol. 13 Journal Issue: 1; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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