Gadgil Himanshu S, Pabst Karen M, Giorgianni Francesco, Umstot Edward S, Desiderio Dominic M, Beranova-Giorgianni Sarka, Gerling Ivan C, Pabst Michael J
Department of Molecular Sciences, The University of Tennessee Health Science Center, Memphis, USA.
Proteomics. 2003 Sep;3(9):1767-80. doi: 10.1002/pmic.200300532.
We performed a proteomic analysis of monocytes primed by lipopolysaccharide (LPS) in vitro, using two-dimensional gels stained with Coomassie blue. We found 16 proteins of approximately 500 detected that either increased or decreased in abundance as a result of priming by LPS (14 with P </= 0.05). The proteins were identified by comparing the masses of their tryptic peptides with those of all known proteins, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and the SWISS-PROT database. Identities were confirmed by matching the sequence of several tryptic peptides, using liquid chromatography electrospray-ionization quadrupole ion trap mass spectrometry. There were increases in the protective enzymes superoxide dismutase and catalase, in four calgranulins, in the cytokine pre-B cell enhancing factor, and in annexin 2, macrophage capping protein, transketolase, pyruvate kinase, and serine/threonine protein kinase 10. Proteins that decreased in abundance were integrin alpha-IIB, protein disulfide isomerase, and platelet-activating factor acetylhydrolase. Many of these altered proteins have interesting functions in inflammation.
我们使用考马斯亮蓝染色的二维凝胶对体外经脂多糖（LPS）致敏的单核细胞进行了蛋白质组学分析。我们发现，在检测到的约500种蛋白质中，有16种蛋白质因LPS致敏而丰度增加或降低（14种P≤0.05）。通过使用基质辅助激光解吸/电离飞行时间质谱和SWISS-PROT数据库，将其胰蛋白酶肽段的质量与所有已知蛋白质的质量进行比较来鉴定这些蛋白质。通过使用液相色谱电喷雾电离四极杆离子阱质谱对几种胰蛋白酶肽段的序列进行匹配来确认其身份。保护性酶超氧化物歧化酶和过氧化氢酶、四种钙粒蛋白、细胞因子前B细胞增强因子以及膜联蛋白2、巨噬细胞封端蛋白、转酮醇酶、丙酮酸激酶和丝氨酸/苏氨酸蛋白激酶10的含量增加。丰度降低的蛋白质有整合素α-IIB、蛋白质二硫键异构酶和血小板活化因子乙酰水解酶。这些改变的蛋白质中有许多在炎症中具有有趣的功能。
