Inhibition of NFATx activation by an oligopeptide: Disrupting the interaction of NFATx with calcineurin
Inhibition of NFATx activation by an oligopeptide: Disrupting the interaction of NFATx with calcineurin
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DOI：
10.4049/jimmunol.167.5.2677
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发表时间：
2001-09-01
影响因子：
4.4
通讯作者：
Arai, N
中科院分区：
文献类型：
--
作者：
Liu, J;Arai, K;Arai, N
钙依赖性磷酸酶钙调神经磷酸酶（CN）调节NFAT的激活和核转位。我们在这里确定了一个新的CN-结合基序的NFAT家族，NFATx的一个成员，和基于这个基序，Pep 3的肽。Pep 3结合CN并与野生型NFATx竞争CN相互作用。Pep 3内的氨基酸突变表明Pep 3的有效功能需要多个氨基酸残基。通过逆转录病毒介导的基因转移在Th克隆中异位表达Pep 3可以选择性地阻断内源性NFATx的核转位，而它对NFAT家族的另一成员NFATp的核转位几乎没有影响。此外，在转染实验中，Pep 3还阻断了B细胞系M12中转染的NFATx的核转位，但不阻断NFATp的核转位，证明了Pep 3对NFATx的特异性抑制。重要的是，由T细胞克隆产生的几种细胞因子被异位Pep 3严重抑制，并且确实，这些细胞因子的产生被野生型NFATx的表达增强。我们的研究结果表明，选择性抑制NFATx激活和细胞因子表达的Pep 3，并提出了一种新的方法来研究每个NFAT家族成员的生物学。这种方法可以提供一个机会，药理学靶向钙离子依赖性信号事件。
Calcium-dependent phosphatase calcineurin (CN) regulates the activation and nuclear translocation of NFAT. We identify here a novel CN-binding motif in one member of the NFAT family, NFATx, and a peptide based on this motif, Pep3. Pep3 binds CN and competes with wild-type NFATx for CN interaction. Amino acid mutations within Pep3 show that multiple amino acid residues are required for the effective functions of Pep3. Ectopic expression of Pep3 in a Th clone via a retrovirus-mediated gene transfer could selectively block the nuclear translocation of endogenous NFATx, whereas it had little effect on the nuclear translocation of another member of the NFAT family, NFATp. Furthermore, in transfection experiments, Pep3 also blocked the nuclear translocation of transfected NFATx, but not NFATp, in the B cell line M12, demonstrating specific inhibition of Pep3 for NFATx. Importantly, several cytokines produced by the T cell clone were severely repressed by ectopic Pep3, and indeed, the production of these cytokines was enhanced by the expression of wild-type NFATx. Our results show selective inhibition of NFATx activation and cytokine expression by Pep3 and suggest a new approach for studying the biology of each NFAT family member. This approach may provide an opportunity for pharmacological targeting of Ca2+-dependent signaling events.