PDBsum entry 4z98

Hydrolase

PDB id: 4z98

Contents

Protein chain

129 a.a.

Ligands

ACT

Waters ×80

PDB id:

4z98

Name:

Hydrolase

Title:

Crystal structure of hen egg white lysozyme using serial x-ray diffraction data collection

Structure:

LysozymE C. Chain: a. Fragment: residues 19-147. Synonym: 1,4-beta-n-acetylmuramidasE C,allergen gal d iv. Ec: 3.2.1.17

Source:

Gallus gallus. Chicken. Organism_taxid: 9031. Tissue: egg white

Resolution:

1.55Å R-factor: 0.209 R-free: 0.246

Authors:

T.D.Murray,A.Y.Lyubimov,C.M.Ogata,M.Uervirojnangkoorn,A.T.Brunger, J.M.Berger

Key ref:

T.D.Murray et al. (2015). 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. PubMed id: 26457423

Date:

10-Apr-15 Release date: 14-Oct-15

Protein chain

P00698  (LYSC_CHICK) -  Lysozyme C from Gallus gallus

Seq: 147 a.a.

Struc: 129 a.a.

Enzyme reactions

• Enzyme class: E.C.3.2.1.17 - lysozyme.
• Reaction: Hydrolysis of the 1,4-beta-linkages between N-acetyl-D-glucosamine and N-acetylmuramic acid in peptidoglycan heteropolymers of the prokaryotes cell walls.

Acta Crystallogr D Biol Crystallogr 71:1987-1997 (2015)

PubMed id: 26457423

A high-transparency, micro-patternable chip for X-ray diffraction analysis of microcrystals under native growth conditions.

T.D.Murray, A.Y.Lyubimov, C.M.Ogata, H.Vo, M.Uervirojnangkoorn, A.T.Brunger, J.M.Berger.

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.