9TOS

Drop on fixed target room temperature serial crystal structure of hen egg white lysozyme, drop on fixed target at DLS, 1.9 s checkerboard control (N-acetyl glucosamine).

Summary for 9TOS

• Entry DOI: 10.2210/pdb9tos/pdb
• Descriptor: Lysozyme C, CHLORIDE ION, SODIUM ION, ... (4 entities in total)
• Functional Keywords: lysozyme, serial, room temperature, hydrolase
• Biological source: Gallus gallus (chicken)
• Total number of polymer chains: 1
• Total formula weight: 14495.96

Authors

Hinchliffe, P.,Kamps, J.A.G.,Aller, P.,Orville, A.M. (deposition date: 2025-12-17, release date: 2026-06-17)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.67 Å)