4Z98

Crystal Structure of Hen Egg White Lysozyme using Serial X-ray Diffraction Data Collection

Summary for 4Z98

• Entry DOI: 10.2210/pdb4z98/pdb
• Descriptor: Lysozyme C, ACETATE ION (3 entities in total)
• Functional Keywords: hydrolase
• Biological source: Gallus gallus (Chicken)
• Cellular location: Secreted: P00698
• Total number of polymer chains: 1
• Total formula weight: 14390.20

Authors

Murray, T.D.,Lyubimov, A.Y.,Ogata, C.M.,Uervirojnangkoorn, M.,Brunger, A.T.,Berger, J.M. (deposition date: 2015-04-10, release date: 2015-10-14, Last modification date: 2024-11-13)

Primary citation

Murray, T.D.,Lyubimov, A.Y.,Ogata, C.M.,Vo, H.,Uervirojnangkoorn, M.,Brunger, A.T.,Berger, J.M.
A high-transparency, micro-patternable chip for X-ray diffraction analysis of microcrystals under native growth conditions.
Acta Crystallogr. D Biol. Crystallogr., 71:1987-1997, 2015

PubMed Abstract: Microcrystals present a significant impediment to the determination of macromolecular structures by X-ray diffraction methods. Although microfocus synchrotron beamlines and X-ray free-electron lasers (XFELs) can enable the collection of interpretable diffraction data from microcrystals, there is a need for efficient methods of harvesting small volumes (<2 µl) of microcrystals grown under common laboratory formats and delivering them to an X-ray beam source under native growth conditions. One approach that shows promise in overcoming the challenges intrinsic to microcrystal analysis is to pair so-called `fixed-target' sample-delivery devices with microbeam-based X-ray diffraction methods. However, to record weak diffraction patterns it is necessary to fabricate devices from X-ray-transparent materials that minimize background scattering. Presented here is the design of a new micro-diffraction device consisting of three layers fabricated from silicon nitride, photoresist and polyimide film. The chip features low X-ray scattering and X-ray absorption properties, and uses a customizable blend of hydrophobic and hydrophilic surface patterns to help localize microcrystals to defined regions. Microcrystals in their native growth conditions can be loaded into the chips with a standard pipette, allowing data collection at room temperature. Diffraction data collected from hen egg-white lysozyme microcrystals (10-15 µm) loaded into the chips yielded a complete, high-resolution (<1.6 Å) data set sufficient to determine a high-quality structure by molecular replacement. The features of the chip allow the rapid and user-friendly analysis of microcrystals grown under virtually any laboratory format at microfocus synchrotron beamlines and XFELs.

PubMed: 26457423
DOI: 10.1107/S1399004715015011

Experimental method

X-RAY DIFFRACTION (1.55 Å)