Smalley David M, Root Karen E, Cho Hyungjun, Ross Mark M, Ley Klaus
Mellon Medical Biomarker Discovery Laboratory, Robert M. Berne Cardiovascular Research Center, Charlottesville, VA 22908-1294, USA.
Thromb Haemost. 2007 Jan;97(1):67-80.
Microparticles (MPs) are small membrane vesicles generated by essentially all cell types. In the plasma, most MPs are derived from platelets, but those from other sources, particularly leukocytes (macrophages, lymphocytes, and neutrophils), endothelial cells, and even smooth muscle cells can be detected and appear to play an important role in normal physiology and various diseases. In previous work we analyzed the proteome of MPs generated from isolated platelets (platelet MPs). Here, we report on a comparative analysis of microparticles isolated from plasma (plasma MPs) versus platelet MP using two complementary methods of comparative analysis. The first method, spectral count analysis, yielded 21 proteins detected in plasma MPs (with a total spectral count of 10 or greater) that were essentially absent in platelet MPs (with a total spectral count of 1 or 0). An additional two proteins (von Willebrand Factor, albumin) were present in both types of MPs but enriched in the plasma MPs. The second method, isotope-coded affinity tag (ICAT) labeling of proteins, supported the spectral count results for the more abundant proteins and provided better relative quantitation of differentially expressed proteins. Proteins present only in the plasma MPs include several associated with apoptosis (CD5-like antigen, galectin 3 binding protein, several complement components), iron transport (transferrin, transferrin receptor, haptoglobin), immune response (complement components, immunoglobulin J and kappa chains), and the coagulation process (protein S, coagulation factor VIII).
微粒(MPs)是由几乎所有细胞类型产生的小膜泡。在血浆中,大多数MPs来源于血小板,但也能检测到来自其他来源的微粒,特别是白细胞(巨噬细胞、淋巴细胞和中性粒细胞)、内皮细胞,甚至平滑肌细胞产生的微粒,它们似乎在正常生理和各种疾病中发挥重要作用。在之前的工作中,我们分析了从分离的血小板中产生的微粒(血小板微粒)的蛋白质组。在此,我们报告了使用两种互补的比较分析方法,对从血浆中分离的微粒(血浆微粒)与血小板微粒进行的比较分析。第一种方法是光谱计数分析,结果显示在血浆微粒中检测到21种蛋白质(总光谱计数为10或更高),而在血小板微粒中基本不存在(总光谱计数为1或0)。另外两种蛋白质(血管性血友病因子、白蛋白)在两种类型的微粒中都存在,但在血浆微粒中更为丰富。第二种方法是对蛋白质进行同位素编码亲和标签(ICAT)标记,该方法支持了对丰度较高蛋白质的光谱计数结果,并为差异表达蛋白质提供了更好的相对定量。仅存在于血浆微粒中的蛋白质包括几种与细胞凋亡相关的蛋白质(CD5样抗原、半乳糖凝集素3结合蛋白、几种补体成分)、铁转运蛋白(转铁蛋白、转铁蛋白受体、触珠蛋白)、免疫反应相关蛋白(补体成分、免疫球蛋白J和κ链)以及凝血过程相关蛋白(蛋白S、凝血因子VIII)。