Jin Ming, Diaz Philip T, Bourgeois Tran, Eng Charis, Marsh Clay B, Wu Haifeng M
Department of Pathology, The Ohio State University, Columbus, OH, USA.
Proteome Sci. 2006 Sep 1;4:16. doi: 10.1186/1477-5956-4-16.
Blood monocytes play a central role in regulating host inflammatory processes through chemotaxis, phagocytosis, and cytokine production. However, the molecular details underlying these diverse functions are not completely understood. Understanding the proteomes of blood monocytes will provide new insights into their biological role in health and diseases.
In this study, monocytes were isolated from five healthy donors. Whole monocyte lysates from each donor were then analyzed by 2D gel electrophoresis, and proteins were detected using Sypro Ruby fluorescence and then examined for phosphoproteomes using ProQ phospho-protein fluorescence dye. Between 1525 and 1769 protein spots on each 2D gel were matched, analyzed, and quantified. Abundant protein spots were then subjected to analysis by mass spectrometry. This report describes the protein identities of 231 monocyte protein spots, which represent 164 distinct proteins and their respective isoforms or subunits. Some of these proteins had not been previously characterized at the protein level in monocytes. Among the 231 protein spots, 19 proteins revealed distinct modification by protein phosphorylation.
The results of this study offer the most detailed monocyte proteomic database to date and provide new perspectives into the study of monocyte biology.
血液单核细胞在通过趋化作用、吞噬作用和细胞因子产生来调节宿主炎症过程中发挥核心作用。然而，这些多样功能背后的分子细节尚未完全明确。了解血液单核细胞的蛋白质组将为其在健康和疾病中的生物学作用提供新的见解。
在本研究中，从五名健康供体中分离出单核细胞。然后对每个供体的全单核细胞裂解物进行二维凝胶电泳分析，使用Sypro Ruby荧光检测蛋白质，接着使用ProQ磷酸化蛋白质荧光染料检测磷酸化蛋白质组。对每张二维凝胶上1525至1769个蛋白质斑点进行匹配、分析和定量。然后对丰富的蛋白质斑点进行质谱分析。本报告描述了231个单核细胞蛋白质斑点的蛋白质身份，这些斑点代表164种不同的蛋白质及其各自的异构体或亚基。其中一些蛋白质此前尚未在单核细胞的蛋白质水平上得到表征。在这231个蛋白质斑点中，有19种蛋白质显示出明显的蛋白质磷酸化修饰。
本研究结果提供了迄今为止最详细的单核细胞蛋白质组数据库，并为单核细胞生物学研究提供了新的视角。
