Isolation and sequence analysis of serine protease cDNAs from mouse cytolytic T lymphocytes
Isolation and sequence analysis of serine protease cDNAs from mouse cytolytic T lymphocytes
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小鼠溶细胞 T 淋巴细胞丝氨酸蛋白酶 cDNA 的分离和序列分析
DOI：
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发表时间：
1988
影响因子：
15.3
通讯作者：
John Ding
中科院分区：
文献类型：
--
作者：
Byoung S. Kwon;Daniel Kestler;Esther Lee;M. Wakulchik;John Ding
通过改进的差异筛选程序从克隆的溶细胞性 T 淋巴细胞 (CTL) 中分离出编码小鼠丝氨酸蛋白酶的三个新 cDNA 克隆（命名为 MCSP-1、MCSP-2 和 MCSP-3）。 MCSP-2和MCSP-3的推定成熟蛋白均由228个氨基酸组成，分子量分别为25,477和25,360。 MCSP-2 和 MCSP-3 预测蛋白的 NH2 末端氨基酸分别与报道的颗粒酶 E 和 F 的氨基酸相同。第三个物种，MCSP-1，与其他两个 cDNA 物种密切相关，但 cDNA NH2 末端部分的大约 30 个氨基酸当量未克隆。形成丝氨酸蛋白酶活性位点的氨基酸在三种预测的蛋白质中非常保守。活性位点口袋残基位于活性位点 Ser184 之前的 6 个残基，在 MCSP-1 中是丙氨酸，在 MCSP-2 中是苏氨酸，在 MCSP-3 中是丝氨酸，表明 MCSP-2 和 MCSP-3 可能都具有胰凝乳蛋白酶样特异性。 MCSP-1 和 MCSP-3 中有 3 个潜在的天冬酰胺连接糖基化位点，MCSP-2 推导的氨基酸序列中有 4 个潜在的天冬酰胺连接糖基化位点。 MCSP-1与其他四种已报道的活性位点口袋残基为丙氨酸的丝氨酸蛋白酶的氨基酸比较表明，MCSP-1与其他分子有显着不同，表明MCSP-1可能是小鼠T细胞丝氨酸蛋白酶家族的新成员。针对 MCSP-1 lacZ 基因融合蛋白的抗体可对 CTL 颗粒进行染色，并在免疫印迹上发生反应，产生 29 和 35-40 kD 的两条不同的颗粒蛋白条带。仅 35-kD 物种带有 [3H]DFP 标签。由于蛋白酶级联可能在溶细胞淋巴细胞激活中发挥关键作用，因此我们分离代表独特丝氨酸酯酶的 cDNA 应有助于研究此类级联过程。
Three new cDNA clones (designated MCSP-1, MCSP-2, and MCSP-3) encoding mouse serine proteases were isolated from cloned cytolytic T lymphocytes (CTL) by a modified differential screening procedure. The putative mature proteins of MCSP-2 and MCSP-3 are each composed of 228 amino acids with molecular weights of 25,477 and 25,360, respectively. NH2-terminal amino acids of MCSP-2- and MCSP-3-predicted proteins were identical to those reported for granzyme E and F, respectively. The third species, MCSP-1, was closely related to the two other cDNA species but approximately 30 amino acids equivalents of the NH2- terminal portion of the cDNA were not cloned. The amino acids forming the active sites of serine proteases were well conserved among the three predicted proteins. The active site pocket residue positioned six residues before the active-site Ser184 is alanine in MCSP-1, threonine in MCSP-2, and serine in MCSP-3, indicating that both MCSP-2 and MCSP-3 may have chymotrypsin-like specificity. There are three potential asparagine-linked glycosylation sites in MCSP-1 and MCSP-3, and four in MCSP-2-deduced amino acid sequences. Amino acid comparison of MCSP-1 with four other reported serine proteases whose active site pocket residue is alanine revealed that MCSP-1 was substantially different from the other molecules, indicating that MCSP-1 may be a new member of mouse T cell serine protease family. Antibodies made against a MCSP-1 lacZ gene fusion protein stain granules of CTL and react on immunoblots with two distinct granule protein bands of 29 and 35-40 kD. Only the 35- kD species labels with [3H]DFP. Since a protease cascade may play a key role in cytolytic lymphocyte activation, our isolation of cDNAs representative of unique serine esterases should help to investigate such a cascade process.
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人类自然杀伤细胞：生物学和病理学方面。
DOI：
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发表时间：
1984
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
Trinchieri,G;Perussia,B
通讯作者：
Perussia,B
影响因子：
4.3
作者：
Redelman,D;Hudig,D
通讯作者：
Hudig,D
DOI：
10.1073/pnas.83.18.7013
发表时间：
1986-09-01
影响因子：
11.1
作者：
MEHRA, V;SWEETSER, D;YOUNG, RA
通讯作者：
YOUNG, RA
DOI：
10.1073/pnas.80.5.1194
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
YOUNG, RA;DAVIS, RW
通讯作者：
DAVIS, RW