CALPAIN EXPRESSION IN LYMPHOID-CELLS - INCREASED MESSENGER-RNA AND PROTEIN-LEVELS AFTER CELL ACTIVATION
CALPAIN EXPRESSION IN LYMPHOID-CELLS - INCREASED MESSENGER-RNA AND PROTEIN-LEVELS AFTER CELL ACTIVATION
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DOI：
10.1074/jbc.270.6.2497
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发表时间：
1995-02-10
影响因子：
4.8
通讯作者：
BANIK, NL
中科院分区：
文献类型：
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作者：
DESHPANDE, RV;GOUST, JM;BANIK, NL
虽然钙蛋白酶普遍存在于人体组织中，并被认为在脱髓鞘中发挥作用，但其活性在静息正常淋巴细胞中非常低。为了确定人类淋巴细胞中钙蛋白酶在mRNA和蛋白水平表达的性质，我们研究了人类T淋巴细胞、B淋巴细胞和单核细胞系以及外周血单核细胞。用佛波醇酯佛波醇肉豆蔻酸酯乙酸酯和钙离子载体A23187刺激细胞导致钙蛋白酶mRNA和蛋白质表达增加。钙蛋白酶mRNA的表达也增加了人T细胞刺激与抗CD 3。RNA和蛋白质的增加之间的分离表明钙蛋白酶可以从细胞中释放出来;随后的实验表明，在细胞外环境中，5，6-二氯-1b-D-呋喃核糖基苯并咪唑（mRNA合成的可逆抑制剂）的存在使钙蛋白酶mRNA水平降低50-67%，蛋白水平降低72- 91%，其去除导致钙蛋白酶mRNA和蛋白质合成的恢复。翻译抑制剂环己酰亚胺使活化的THP-1细胞中钙蛋白酶蛋白水平降低77-81%，钙蛋白酶mRNA水平降低96%。干扰素-γ诱导U-937和THP-1细胞钙蛋白酶mRNA和蛋白表达。地塞米松增加THP-1细胞mRNA的表达。我们的结果表明，淋巴细胞的激活导致钙蛋白酶的从头合成和分泌。
Although calpain is ubiquitously present in human tissues and is thought to play a role in demyelination, its activity is very low in resting normal lymphocytes. To determine the nature of calpain expression at the mRNA and protein levels in human lymphoid cells, we studied human T lymphocytic, B lymphocytic, and monocytic Lines as well as peripheral blood mononuclear cells. Stimulation of cells with the phorbol ester phorbol myristate acetate and the calcium ionophore A23187 resulted in increased calpain mRNA and protein expression. Calpain mRNA expression is also increased in human T cells stimulated with anti-CD3. A dissociation between the increases of RNA and protein suggested that calpain could be released from the cells; the subsequent experiments showed its presence in the extracellular environment, 5,6-Dichloro-1b-D-ribofuranosylbenzimidazoIe, a reversible inhibitor of mRNA synthesis, reduced calpain mRNA levels by 50-67% and protein levels by 72-91%, Its removal resulted in resumption of both calpain mRNA and protein synthesis. Cycloheximide, a translational inhibitor, reduced calpain protein levels by 77-81% and calpain mRNA levels by 96% in activated THP-1 cells. Interferon-gamma induced calpain mRNA and protein in U-937 and THP-1 cells. Dexamethasone increased mRNA expression in THP-1 cells, Our results indicate that activation of lymphoid cells results in de novo synthesis and secretion of calpain.