Characterization of GMP-17, a granule membrane protein that moves to the plasma membrane of natural killer cells following target cell recognition
Characterization of GMP-17, a granule membrane protein that moves to the plasma membrane of natural killer cells following target cell recognition
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DOI：
10.1073/pnas.93.2.685
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发表时间：
1996-01-23
影响因子：
11.1
通讯作者：
Anderson, P
中科院分区：
文献类型：
--
作者：
Medley, QG;Kedersha, N;Anderson, P
细胞毒性淋巴细胞的特征在于其包含在靶细胞识别后与质膜融合的细胞质颗粒。我们以前鉴定了一种称为p15-TIA-1的细胞毒性颗粒膜蛋白，其与称为p40-TIA-1的RNA识别基序（RRM）型RNA结合蛋白免疫化学相关。尽管有人认为p15-TIA-1可能通过蛋白水解衍生自p40-TIA-1，通过使用单克隆抗体（mAb）2G 9从自然杀伤（NK）细胞系免疫亲和纯化的p15-TIA-1的N-末端氨基酸测序显示，p15-TIA-1在NK细胞系中的N-末端氨基酸序列与p15-TIA-1的N-末端氨基酸序列一致。1的氨基酸序列与NK细胞和粒细胞集落刺激因子处理的单个核细胞中的NKG 7和GIG-1的cDNA序列完全相同。表位分析表明，mAb 2G 9识别p15-TIA-1和p40-TIA-1的C末端，p15-TIA-1/NKG 7/GIG-1的推导氨基酸序列预测该蛋白具有四个跨膜结构域，并且免疫电子显微镜将内源性蛋白定位于NK细胞中的细胞毒性颗粒的膜。鉴于其亚细胞定位，我们建议将该蛋白质重新命名为GMP-17，即17 kDa的颗粒膜蛋白。新鲜分离的NK细胞的免疫荧光显微镜检查证实了这种颗粒定位。靶细胞诱导的NK细胞脱粒导致GMP-17从颗粒易位到质膜，提示GMP-17在调节淋巴细胞和嗜中性粒细胞的效应子功能中的可能作用。
Cytotoxic lymphocytes are characterized by their inclusion of cytoplasmic granules that fuse with the plasma membrane following target cell recognition. We previously identified a cytotoxic granule membrane protein designated p15-TIA-1 that is immunochemically related to an RNA-recognition motif (RRM)-type RNA-binding protein designated p40-TIA-1, Although it was suggested that p15-TIA-1 might be derived from p40-TIA-1 by proteolysis, N-terminal amino acid sequencing of p15-TIA-1 immunoaffinity purified from a natural killer (NK) cell line by using monoclonal antibody (mAb) 2G9 revealed that p15-TIA-1 is identical to the deduced amino acid sequence of NKG7 and GIG-1, cDNAs isolated from NK cells and granulocyte-colony-stimulating factor-treated mononuclear cells, respectively, Epitope mapping revealed that mAb 2G9 recognizes the C terminus of p15-TIA-1 and p40-TIA-1, The deduced amino acid sequence of p15-TIA-1/NKG7/GIG-1 predicts that the protein possesses four transmembrane domains, and immune-electron microscopy localizes the endogenous protein to the membranes of cytotoxic granules in NK cells, Given its subcellular localization, we propose to rename this protein GMP-17, for granule membrane protein of 17 kDa, Immunofluorescence microscopy of freshly isolated NK cells confirms this granular localization, Target cell-induced NK cell degranulation results in translocation of GMP-17 from granules to the plasma membrane, suggesting a possible role for GMP-17 in regulating the effector function of lymphocytes and neutrophils.