Direct Demonstration of NFATp Dephosphorylation and Nuclear Localization in Activated HT-2 Cells Using a Specific NFATp Polyclonal Antibody (*)
Direct Demonstration of NFATp Dephosphorylation and Nuclear Localization in Activated HT-2 Cells Using a Specific NFATp Polyclonal Antibody (*)
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使用特定的 NFATp 多克隆抗体直接演示激活的 HT-2 细胞中的 NFATp 去磷酸化和核定位 (*)
DOI：
--
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发表时间：
1995
影响因子：
4.8
通讯作者：
K. Leach
中科院分区：
文献类型：
--
作者：
V. Ruff;K. Leach
活化T细胞核因子(NFAT)调节多种细胞因子基因的转录，并在T细胞活化后激发NFAT DNA结合活性。一些证据表明，NFAT是钙调神经磷酸酶(一种丝氨酸/苏氨酸磷酸酶)的底物。用抗小鼠NFATpp的多克隆抗体对小鼠不同组织进行免疫印迹分析表明，110-130 kDa的NFATp蛋白在胸腺和脾中高表达。用钙调神经磷酸酶处理未处理的HT-2细胞中免疫沉淀的NFATp，导致NFATp去磷酸化，表明NFATp是钙调神经磷酸酶的体外底物。从~(32)P标记的HT-2细胞中免疫沉淀的NFATP以大约120 kDa的蛋白迁移到细胞的胞浆中。用离子霉素处理细胞后，NFATp的相对分子质量降低，32P丢失，与NFATp去磷酸化一致。NFATp的去磷酸化伴随着蛋白质定位到核组分。用钙调神经磷酸酶抑制剂FK506预先孵育细胞可以阻断这两个事件，这与NFATP是细胞中钙调神经磷酸酶底物的假设一致。
Nuclear factor of activated T cells (NFAT) regulates transcription of a number of cytokine genes, and NFAT DNA binding activity is stimulated following T cell activation. Several lines of evidence have suggested that NFAT is a substrate for calcineurin, a serine/threonine phosphatase. Using a polyclonal antibody to murine NFATpp, Western blot analysis of various mouse tissues demonstrated that the 110-130-kDa NFATp protein was highly expressed in thymus and spleen. Treatment of immunoprecipitated NFATp from untreated HT-2 cells with calcineurin resulted in the dephosphorylation of NFATp, demonstrating that NFATp is an in vitro substrate for calcineurin. NFATp immunoprecipitated from 32P-labeled HT-2 cells migrated as an approximately 120-kDa protein that was localized to the cytosol of the cells. Treatment of the cells with ionomycin resulted in a decrease in the molecular weight of NFATp and a loss of 32P, consistent with NFATp dephosphorylation. The dephosphorylation of NFATp was accompanied by localization of the protein to the nuclear fraction. Both of these events were blocked by preincubation of the cells with FK506, a calcineurin inhibitor, consistent with the hypothesis that NFATp is a calcineurin substrate in cells.
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DOI：
10.1016/s0021-9258(18)53757-1
发表时间：
1993-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
P. Mccaffrey;B. Perrino;T. Soderling;A. Rao
通讯作者：
P. Mccaffrey;B. Perrino;T. Soderling;A. Rao
DARPP-32（Mr 32,000 多巴胺和 cAMP 调节磷蛋白）的合成肽类似物，是蛋白磷酸酶 1 的抑制剂。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
HemmingsJr,HC;Nairn,AC;Elliott,JI;Greengard,P
通讯作者：
Greengard,P
DOI：
10.1016/s0021-9258(18)46911-6
发表时间：
1994-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. M. Ho;J. Jain;A. Rao;P. Hogan
通讯作者：
A. M. Ho;J. Jain;A. Rao;P. Hogan
影响因子：
2.9
作者：
LIU, J;ALBERS, MW;SCHREIBER, SL
通讯作者：
SCHREIBER, SL
影响因子：
4.4
作者：
J. Jain;Zoe Miner;Anjana Rao
通讯作者：
J. Jain;Zoe Miner;Anjana Rao