Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA.
Rapid Commun Mass Spectrom. 2010 Jan;24(1):11-4. doi: 10.1002/rcm.4349.
Bacteriophage (phage) proteins have been analyzed previously with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). However, analysis of phage major capsid proteins (MCPs) has been limited by the ability to reproducibly generate ions from MCP monomers. While the acidic conditions of MALDI-TOF MS sample preparation have been shown to aid in disassembly of some phage capsids, many require further treatment to successfully liberate MCP monomers. The findings presented here suggest that beta-mercaptoethanol reduction of the disulfide bonds linking phage MCPs prior to mass spectrometric analysis results in significantly increased MALDI-TOF MS sensitivity and reproducibility of Yersinia pestis-specific phage protein profiles.
先前已经使用基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF MS）分析了噬菌体（phage）蛋白。然而，由于能够从 MCP 单体中重现地产生离子，因此对噬菌体主要衣壳蛋白（MCP）的分析受到了限制。虽然 MALDI-TOF MS 样品制备的酸性条件已被证明有助于某些噬菌体衣壳的解体，但许多条件需要进一步处理才能成功释放 MCP 单体。这里提出的发现表明，在质谱分析之前，用β-巯基乙醇还原将噬菌体 MCPs 连接的二硫键，可显著提高鼠疫耶尔森氏菌特异性噬菌体蛋白图谱的 MALDI-TOF MS 灵敏度和重现性。
