Lee Soo Jae, Kim Kyun-Hwan, Park Ji Sook, Jung Jin Woo, Kim Young Hwan, Kim Sang Kyung, Kim Wan-Seok, Goh Hyun-gyung, Kim Soo-hyun, Yoo Jung-Sun, Kim Dong-Wook, Kim Kwang Pyo
Department of Molecular Biotechnology, Institute of Biomedical Science and Technology, Konkuk University, Seoul 143-701, Republic of Korea.
Biochem Biophys Res Commun. 2007 Jun 8;357(3):620-6. doi: 10.1016/j.bbrc.2007.03.191. Epub 2007 Apr 9.
This study was designed to identify the cell surface protein markers that can differentiate between chronic myeloid leukemia (CML) and acute promyelocytic leukemia cells (APL). The differentially expressed plasma membrane proteins were analyzed between CML cell line (K562) and APL cell line (NB4) using the comparative proteomic approach. The cell membrane proteins were enriched by labeling with a membrane-impermeable biotinylation reagent, sulfo-NHS-SS-Biotin, and subjected to liquid chromatography tandem mass spectrometry (LC-MS/MS). By comparative proteomic analysis of K562 and NB4 cells, we identified 25 membrane and 14 membrane-associated proteins. The result of LC-MS/MS combined with chemical tagging method was validated by confirming the expression and localization of one of the differentially expressed plasma membrane proteins, CD43, by FACS and confocal microscopy. Our results indicate that CD43 could be a potential candidate for differentiating CML from APL.
本研究旨在鉴定可区分慢性髓性白血病（CML）细胞和急性早幼粒细胞白血病细胞（APL）的细胞表面蛋白标志物。采用比较蛋白质组学方法分析了CML细胞系（K562）和APL细胞系（NB4）之间差异表达的质膜蛋白。通过用一种膜不可渗透的生物素化试剂磺基-NHS-SS-生物素进行标记来富集细胞膜蛋白，然后进行液相色谱串联质谱分析（LC-MS/MS）。通过对K562和NB4细胞进行比较蛋白质组学分析，我们鉴定出了25种膜蛋白和14种膜相关蛋白。通过流式细胞术（FACS）和共聚焦显微镜确认差异表达的质膜蛋白之一CD43的表达和定位，验证了LC-MS/MS结合化学标记方法的结果。我们的结果表明，CD43可能是区分CML和APL的潜在候选标志物。
