Sasaki Kazuki, Tsuchiya Takashi, Osaki Tsukasa
National Cerebral and Cardiovascular Center, Osaka, Japan.
Methods Mol Biol. 2018;1719:51-58. doi: 10.1007/978-1-4939-7537-2_3.
Media conditioned by cultured cells represent an excellent source rich in endogenous peptides. Unbiased mass spectrometric analysis of the constituent peptides provides an opportunity to look into proteolytic events such as bioactive peptide processing, membrane protein ectodomain shedding, or even regulated intramembrane proteolysis. If conducted on a large scale, peptidomics has the potential to pinpoint primary cleavage sites. Here a method is described for isolating peptides from cultured cell conditioned media before mass spectrometry analysis.
由培养细胞条件化的培养基是富含内源性肽的优质来源。对组成肽进行无偏倚的质谱分析，为研究蛋白水解事件提供了机会，例如生物活性肽加工、膜蛋白胞外域脱落，甚至是调节性膜内蛋白水解。如果大规模开展，肽组学有潜力确定主要切割位点。本文描述了一种在质谱分析前从培养细胞条件培养基中分离肽的方法。
