9TOP

Contaminated room temperature serial crystal structure of CTX-M-15 class A beta-lactamase, drop on fixed target at DLS, 2.6 s 'control' contaminated with avibactam

9TOP の概要

• エントリーDOI: 10.2210/pdb9top/pdb
• 分子名称: Beta-lactamase, (2S,5R)-1-formyl-5-[(sulfooxy)amino]piperidine-2-carboxamide, (2S,5R)-7-oxo-6-(sulfooxy)-1,6-diazabicyclo[3.2.1]octane-2-carboxamide, ... (6 entities in total)
• 機能のキーワード: antibiotic resistance, beta-lactamase, inhibitor, dynamics, tr-ssx, antimicrobial protein
• 由来する生物種: Klebsiella pneumoniae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 28906.93

構造登録者

Parkinson, L.,Tooke, C.L.,Hinchliffe, P.,Beer, M.,Spencer, J. (登録日: 2025-12-17, 公開日: 2026-06-17)

主引用文献

Kamps, J.J.A.G.,Hinchliffe, P.,Glerup, J.,Freeman, E.I.,Lang, P.A.,Tooke, C.L.,Beer, M.,Parkinson, L.,Gu, D.H.,Park, S.,Devenish, N.,Zhou, T.,Shilova, A.,Kaur, S.,Rabe, P.,Schofield, C.J.,Spencer, J.,Park, J.,Owen, R.L.,Orville, A.M.,Aller, P.
Drop-on-fixed-target reaction initiation approach for serial and time-resolved crystallography.
Iucrj, 2026

PubMed Abstract: We describe the design and implementation of a drop-on-fixed-target method for time-resolved serial crystallography at both synchrotron and XFEL facilities. A piezoelectric droplet dispensing pipette is employed for addition of picolitre volume aqueous droplets (∼40-90 pl; ∼40-55 µm diameter sphere), containing (co-)substrate(s) or ligand(s), onto enzyme microcrystals previously loaded into the trapezoidal wells of an etched crystalline silicon fixed-target chip containing 25 600 wells in a high-density, square grid with 125 µm centre-to-centre well spacing. These features demand exquisite accuracy and thereby constrain motion controls to enable robust time-resolved crystallographic studies. The system was tested with three enzyme systems, comprising lysozyme and two β-lactamases, CTX-M-15 and AmpC. Mitigation strategies for cross-well contamination, including the implementation of interleaved controls, are described; the overall performance of the system at synchrotron and X-ray free-electron laser facilities was evaluated. This drop-on-fixed-target method is a reliable framework for time-resolved crystallography and will improve the consistency of measurements across facilities.

PubMed: 42246252
DOI: 10.1107/S2052252526003489

実験手法

X-RAY DIFFRACTION (1.8 Å)