Proteomic analysis of macrophages: a new way to identify novel cell-surface antigens.
Macrophages are involved in many important biological processes and membrane proteins are the key effector molecules for their function. However, membrane proteins are difficult to analyze by 2-DE based methods because of their intrinsic tendency to self-aggregate during the first dimension separation (IEF). To circumvent this, we combined one-dimensional SDS-PAGE with capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this technique, we identified 458 GO annotated membrane proteins with extremely high confidence, including most known markers of peritoneal macrophages (e.g., CD11b, F4/80, CD14, CD18, CD86, CD44, CD16 and Toll-like receptor). Thirteen other CD antigens (CD243, CD98, CD107a, CD107b, CD36, CD97, CD205, CD206, CD180, CD191, CD300, CD45and CD29), and 18 Ras-related small GTPases were also identified. In addition to those known macrophage membrane proteins, a significant number of novel proteins have also been identified. This research not only provides a technique to study membrane proteins, but also a valuable dataset of macrophage antigens, thus providing better understanding of the functional mechanisms of macrophages in many biological processes.
Zhang
Lingbing
L
Department of Immunology, College of Basic Medicine, Jilin University, Changchun 130021, China.
Lun
Yanni
Y
Yan
Dongmei
D
Yu
Leyang
L
Ma
Wei
W
Du
Bairong
B
Zhu
Xun
X
D000818:Animals; D015703:Antigens, CD / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis*; D053801:Capillary Electrochromatography / Q000379:methods; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005260:Female; D005434:Flow Cytometry; D017737:Macrophages, Peritoneal / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D020559:Monomeric GTP-Binding Proteins / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
D015703:Antigens, CD; D000954:Antigens, Surface; D020559:Monomeric GTP-Binding Proteins