CLONING OF A CDNA FOR A-T CELL-SPECIFIC SERINE PROTEASE FROM A CYTOTOXIC LYMPHOCYTE-T
CLONING OF A CDNA FOR A-T CELL-SPECIFIC SERINE PROTEASE FROM A CYTOTOXIC LYMPHOCYTE-T
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DOI：
10.1126/science.2422755
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发表时间：
1986-05-16
期刊：
影响因子：
56.9
通讯作者：
WEISSMAN, IL
中科院分区：
文献类型：
--
作者：
GERSHENFELD, HK;WEISSMAN, IL
通过 RNA 杂交竞争方案，从鼠细胞毒性 T 淋巴细胞互补 DNA 文库中分离出克隆，编码了一种新的丝氨酸蛋白酶。在细胞毒性 T 淋巴细胞、裸鼠脾细胞、大鼠自然杀伤细胞白血病以及八个 T 辅助细胞克隆中的两个（均为细胞毒性）中检测到了互补转录物，但在正常小鼠肾、肝、脾或胸腺中以及在几种测试的 T 和 B 细胞肿瘤中均未检测到。用刀豆球蛋白 A 加白细胞介素 2 激活 T 细胞，诱导脾细胞表达该基因，其动力学与获得细胞溶解能力相关。该基因的核苷酸序列编码大约25,700道尔顿的氨基酸序列，与丝氨酸蛋白酶家族的成员具有25%至35%的同一性。活性位点“电荷中继”残基（胰凝乳蛋白酶编号系统的 His57、Asp102 和 Ser195）以及胰蛋白酶特异性 Asp（胰蛋白酶中的位置 189）是保守的。 Southern印迹分析表明该基因在人类、小鼠和鸡中是保守的。这种丝氨酸蛋白酶可能在淋巴细胞裂解和“裂解级联”中发挥作用。
A new serine protease was encoded by a clone isolated from a murine cytotoxic T-lymphocyte complementary DNA library by an RNA-hybridization competition protocol. Complementary transcripts were detected in cytotoxic T lymphocytes, spleen cells from nude mice, a rat natural killer cell leukemia, and in two of eight T-helper clones (both cytotoxic), but not in normal mouse kidney, liver, spleen, or thymus, nor in several tested T- and B-cell tumors. T-cell activation with concanavalin A plus interleukin-2 induced spleen cells to express this gene with kinetics correlating with the acquisition of cytolytic capacity. The nucleotide sequence of this gene encoded an amino acid sequence of approximately 25,700 daltons, with 25 to 35 percent identity to members of the serine protease family. The active site "charge-relay" residues (His57, Asp102, and Ser195 of the chymotrypsin numbering system) are conserved, as well as the trypsin-specific Asp (position 189 in trypsin). A Southern blot analysis indicated that this gene is conserved in humans, mouse, and chicken. This serine protease may have a role in lymphocyte lysis and a "lytic cascade".