Swatkoski Stephen, Russell Scott, Edwards Nathan, Fenselau Catherine
Department of Chemistry & Biochemistry and Center for Bioinformatics and Computational Biology, University of Maryland, College Park, Maryland 20742, USA.
Anal Chem. 2007 Jan 15;79(2):654-8. doi: 10.1021/ac061493e.
A nonenzymatic proteomics strategy is applied to the rapid identification of viruses. The approach provides solubilization and subsequent digestion of viral coat proteins in under 30 s. Acid digestions were carried out using a laboratory-quality microwave system equipped with temperature, pressure, and power controls, which allowed for precise optimization of experimental parameters. Under optimal conditions, this method provides an efficient alternative to traditional enzymatic digestion-based methods for virus identification. Following rapid microwave heating of a suspension of a model virus, RNA bacteriophage MS2, 13 chemical digestion products were detected in parallel with the coat protein precursor using MALDI-TOF MS. Because of the high sequence coverage obtained, the bacteriophage MS2 coat protein was identified with high confidence and the specificity of the identification allowed for the discrimination between bacteriophage MS2 and other closely related RNA bacteriophages.
一种非酶蛋白质组学策略被应用于病毒的快速鉴定。该方法能在30秒内实现病毒衣壳蛋白的溶解及后续消化。酸消化是使用配备温度、压力和功率控制装置的实验室级微波系统进行的，这使得实验参数能够得到精确优化。在最佳条件下，该方法为基于传统酶消化的病毒鉴定方法提供了一种有效的替代方案。对模型病毒RNA噬菌体MS2的悬浮液进行快速微波加热后，使用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）同时检测到了13种化学消化产物以及衣壳蛋白前体。由于获得了高序列覆盖率，噬菌体MS2衣壳蛋白得到了高度可靠的鉴定，并且鉴定的特异性使得能够区分噬菌体MS2和其他密切相关的RNA噬菌体。
