5LA8

Room temperature X-ray diffraction of tetragonal HEWL. Third data set (0.93 MGy)

Summary for 5LA8

• Entry DOI: 10.2210/pdb5la8/pdb
• Descriptor: Lysozyme C, CHLORIDE ION, ACETATE ION, ... (4 entities in total)
• Functional Keywords: room temperature, hydrolase
• Biological source: Gallus gallus (Chicken)
• Cellular location: Secreted: P00698
• Total number of polymer chains: 1
• Total formula weight: 14496.56

Authors

Castellvi, A.,Juanhuix, J. (deposition date: 2016-06-13, release date: 2016-12-21, Last modification date: 2024-10-23)

Primary citation

Crosas, E.,Castellvi, A.,Crespo, I.,Fulla, D.,Gil-Ortiz, F.,Fuertes, G.,Kamma-Lorger, C.S.,Malfois, M.,Aranda, M.A.,Juanhuix, J.
Uridine as a new scavenger for synchrotron-based structural biology techniques.
J Synchrotron Radiat, 24:53-62, 2017

PubMed Abstract: Macromolecular crystallography (MX) and small-angle X-ray scattering (SAXS) studies on proteins at synchrotron light sources are commonly limited by the structural damage produced by the intense X-ray beam. Several effects, such as aggregation in protein solutions and global and site-specific damage in crystals, reduce the data quality or even introduce artefacts that can result in a biologically misguiding structure. One strategy to reduce these negative effects is the inclusion of an additive in the buffer solution to act as a free radical scavenger. Here the properties of uridine as a scavenger for both SAXS and MX experiments on lysozyme at room temperature are examined. In MX experiments, upon addition of uridine at 1 M, the critical dose D is increased by a factor of ∼1.7, a value similar to that obtained in the presence of the most commonly used scavengers such as ascorbate and sodium nitrate. Other figures of merit to assess radiation damage show a similar trend. In SAXS experiments, the scavenging effect of 40 mM uridine is similar to that of 5% v/v glycerol, and greater than 2 mM DTT and 1 mM ascorbic acid. In all cases, the protective effect of uridine is proportional to its concentration.

PubMed: 28009546
DOI: 10.1107/S1600577516018452

Experimental method

X-RAY DIFFRACTION (2 Å)