Uridine as a new scavenger for synchrotron-based structural biology techniques
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- 作者：Eva Crosas ; Albert Castellvi ; Isidro Crespo ; Daniel Fulla ; Fernando Gil-Ortiz ; Gustavo Fuertes ; Christina S. Kamma-Lorger ; Marc Malfois ; Miguel A. G. Aranda and Jordi Juanhuix
- 关键词：radiation damage ; reactive oxygen species ; free radical scavenger ; macromolecular crystallography methods ; SAXS methods
- 刊名：Journal of Synchrotron Radiation
- 出版年：2017
- 出版时间：January 2017
- 年：2017
- 卷：24
- 期：1
- 页码：53-62
- 全文大小：948K
- ISSN：1600-5775
文摘
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 D1/2 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.