7UHJ

Time-Resolved Structure of Metallo Beta-Lactamase L1 in a Complex with Non-Hydrolyzed Moxalactam (60 ms Snapshot)

Summary for 7UHJ

• Entry DOI: 10.2210/pdb7uhj/pdb
• Related: 7L91
• Descriptor: Putative metallo-beta-lactamase l1 (Beta-lactamase type ii) (Ec 3.5.2.6) (Penicillinase), ZINC ION, (1R,6R,7R)-7-[(2R)-2-carboxypropanamido]-7-methoxy-3-methyl-8-oxo-5-oxa-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, ... (4 entities in total)
• Functional Keywords: metallo beta lactamase, moxalactam, serial crystallography, structural genomics, center for structural genomics of infectious diseases, hydrolase, csgid
• Biological source: Stenotrophomonas maltophilia K279a
• Total number of polymer chains: 1
• Total formula weight: 29703.03

Authors

Wilamowski, M.,Kim, Y.,Sherrell, D.A.,Lavens, A.,Henning, R.,Maltseva, N.,Endres, M.,Babnigg, G.,Srajer, V.,Joachimiak, A.,Center for Structural Genomics of Infectious Diseases (CSGID) (deposition date: 2022-03-27, release date: 2022-04-13, Last modification date: 2023-10-25)

Primary citation

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.
Time-resolved beta-lactam cleavage by L1 metallo-beta-lactamase.
Nat Commun, 13:7379-7379, 2022

PubMed 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 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 ions and the active site residues. Mechanistic details captured here can be generalized to other MBLs.

PubMed: 36450742
DOI: 10.1038/s41467-022-35029-3

Experimental method

X-RAY DIFFRACTION (2.2 Å)