5LAF

Room temperature X-ray diffraction of tetragonal HEWL with 1M of uridine. First data set (0.31 MGy)

5LAF の概要

• エントリーDOI: 10.2210/pdb5laf/pdb
• 分子名称: Lysozyme C, CHLORIDE ION, ACETATE ION, ... (4 entities in total)
• 機能のキーワード: room temperature, hydrolase
• 由来する生物種: Gallus gallus (Chicken)
• 細胞内の位置: Secreted: P00698
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 14496.56

構造登録者

Castellvi, A.,Juanhuix, J. (登録日: 2016-06-14, 公開日: 2016-12-21, 最終更新日: 2024-11-13)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.5 Å)