Proteolysis in mixed organic-aqueous solvent systems: Applications for peptide mass mapping using mass spectrometry
Proteolysis in mixed organic-aqueous solvent systems: Applications for peptide mass mapping using mass spectrometry
复制标题
DOI：
10.1021/ac001332p
复制
发表时间：
2001-06-01
影响因子：
7.4
通讯作者：
Russell, DH
中科院分区：
文献类型：
--
作者：
Russell, WK;Park, ZY;Russell, DH
蛋白质消化的速度对使用质谱法进行高通量蛋白质鉴定施加了显著的限制。在本报告中，我们证明了蛋白质在有机溶剂（如甲醇、丙酮、2-丙醇和乙腈）的存在下很容易被胰蛋白酶消化。蛋白质在有机溶剂中的消化速率，如质谱中消化片段离子的丰度所示，相对于水溶液增加。此外，在有机溶剂的存在下，氨基酸覆盖在所分析的蛋白质中增加，并且抗蛋白质水解的蛋白质很容易被消化。例如，从肌红蛋白的胰蛋白酶消化获得68%的氨基酸序列覆盖率
The rate of protein digestion imposes significant limitations on high-throughput protein identification using mass spectrometry. In this report, we demonstrate that proteins are readily digested by trypsin in the presence of organic solvents such as methanol, acetone, 2-propanol, and acetonitrile, The rates of protein digestion in organic solvents, as indicated by the abundances of digest fragment ions in the mass spectrum, are increased relative to aqueous solution. In addition, amino acid coverage for the analyzed proteins increases in the presence of the organic solvents, and proteins that are resistant to proteolysis are readily digested. For example, a 68% amino acid sequence coverage was attained from a tryptic digest of myoglobin in