Protein Profiling of Plasma Membranes Defines Aberrant Signaling Pathways in Mantle Cell Lymphoma
Protein Profiling of Plasma Membranes Defines Aberrant Signaling Pathways in Mantle Cell Lymphoma
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DOI：
10.1074/mcp.m800515-mcp200
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发表时间：
2009-07-01
影响因子：
7
通讯作者：
Cain, Kelvin
中科院分区：
文献类型：
--
作者：
Boyd, Robert S.;Jukes-Jones, Rebekah;Cain, Kelvin
我们使用鸟枪蛋白质组学鉴定从白血病期套细胞淋巴瘤（MCL）患者的B细胞中纯化的质膜和脂筏蛋白。生物信息学鉴定了111种跨膜蛋白，其中一些蛋白在原发性MCL病例、MCL衍生的细胞系和正常B细胞中使用RT-PCR和Western印迹进行了分析。与正常B细胞相比，几种跨膜蛋白，包括CD 27、CD 70和CD 31（PECAM-1）过表达。在5名MCL患者中的3名中，CD 70上调（> 10倍）沿着其同源受体CD 27在5名患者中上调（4-9倍），表明MCL细胞可能通过该信号传导途径经历自分泌刺激。在质膜中也检测到激活的钙蛋白酶I和蛋白激酶C β II，表明这些蛋白质在MCL中具有组成性活性。蛋白激酶C β II与脂筏相关，鸟枪蛋白质组学/蛋白质分析显示，关键的脂筏蛋白，raftlin（5例患者中的4例）和CSK（C-末端Src激酶）结合蛋白（Cbp）/与鞘糖脂富集微结构域（PAG）相关的磷蛋白（4例患者中的4例）在MCL中下调。其他已知的脂筏蛋白，如林恩激酶和flotillin 1的水平与正常B细胞相似。然而，5-脂氧合酶（5-LO），在白三烯生物合成的关键酶，与脂筏，并上调MCL与正常B细胞相比，约7倍。5-LO活性抑制剂（AA 861）和5-LO激活蛋白（FLAP）（MK 886，其激活酶）诱导MCL细胞系和原代慢性淋巴细胞白血病细胞凋亡，表明白三烯生物合成途径在MCL和其它B细胞恶性肿瘤中起重要作用。因此，使用鸟枪蛋白质组学和mRNA和蛋白质表达谱，我们确定了一个子集的已知和未知的跨膜蛋白异常表达MCL质膜。这些蛋白质可能在疾病的病理学中起作用，并且是MCL的潜在治疗靶点。Molecular & Cellular Proteomics 8：1501-1515，2009.
We used shotgun proteomics to identify plasma membrane and lipid raft proteins purified from B cells obtained from mantle cell lymphoma (MCL) patients in leukemic phase. Bioinformatics identified 111 transmembrane proteins, some of which were profiled in primary MCL cases, MCL-derived cell lines, and normal B cells using RT-PCR and Western blotting. Several transmembrane proteins, including CD27, CD70, and CD31 (PECAM-1), were overexpressed when compared with normal B cells. CD70 was up-regulated (> 10-fold) in three of five MCL patients along with its cognate receptor CD27, which was up-regulated (4-9-fold) in five of five patients, suggesting that MCL cells may undergo autocrine stimulation via this signaling pathway. Activated calpain I and protein kinase C beta II were also detected in the plasma membranes, suggesting that these proteins are constitutively active in MCL. Protein kinase C beta II has been associated with lipid rafts, and shotgun proteomics/ protein profiling revealed that key lipid raft proteins, raftlin (four of five patients) and CSK (C-terminal Src kinase)-binding protein (Cbp)/phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG) (four of four patients) were down-regulated in MCL. Levels of other known lipid raft proteins, such as Lyn kinase and flotillin 1, were similar to normal B cells. However, 5-lipoxygenase (5-LO), a key enzyme in leukotriene biosynthesis, was associated with lipid rafts and was up-regulated similar to 7-fold in MCL compared with normal B cells. Significantly inhibitors of 5-LO activity (AA861) and 5-LO-activating protein (FLAP) (MK886, its activating enzyme) induced apoptosis in MCL cell lines and primary chronic lymphocytic leukemia cells, indicating an important role for the leukotriene biosynthetic pathway in MCL and other B cell malignancies. Thus, using shotgun proteomics and mRNA and protein expression profiling we identified a subset of known and unknown transmembrane proteins with aberrant expression in MCL plasma membranes. These proteins may play a role in the pathology of the disease and are potential therapeutic targets in MCL. Molecular & Cellular Proteomics 8: 1501-1515, 2009.