Early phosphorylation kinetics of proteins involved in proximal TCR-mediated signaling pathways
Early phosphorylation kinetics of proteins involved in proximal TCR-mediated signaling pathways
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DOI：
10.4049/jimmunol.175.4.2449
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发表时间：
2005-08-15
影响因子：
4.4
通讯作者：
Samelson, LE
中科院分区：
文献类型：
--
作者：
Houtman, JCD;Houghtling, RA;Samelson, LE
通过刺激TCR活化T细胞在适应性免疫应答中起着中心作用。尽管对TCR刺激的信号通路了解很多，但我们对早期激活过程中的动力学和事件顺序以及参与这些近端信号通路的激酶的体内特异性的了解仍然存在差距。这些信息不仅对理解T细胞功能重要的信号通路的激活很重要，而且对药物靶点和基于计算机的分子模型的开发也很重要。在这项研究中，磷酸化特异性抗体针对信号蛋白上的各个位点，用于研究参与近端TCR诱导途径的蛋白质的早期磷酸化动力学。这些研究表明，用于活化T细胞酪氨酸的接头具有显著不同的磷酸化动力学，并且与其他蛋白质相比，含有Src同源2结构域的76 kDa白细胞蛋白具有快速、瞬时的磷酸化动力学。此外，我们提供的证据表明，ZAP-70是LAT酪氨酸191的主要体内激酶，Itk在磷脂酶C-γ 1上酪氨酸783的磷酸化中发挥作用。总的来说，这些研究对T细胞活化至关重要的早期TCR介导的信号传导事件的序列、动力学和特异性提供了新的见解。
Activation of T cells via the stimulation of the TCR plays a central role in the adaptive immunological response. Although much is known about TCR-stimulated signaling pathways, there are still gaps in our knowledge about the kinetics and sequence of events during early activation and about the in vivo specificity of kinases involved in these proximal signaling pathways. This information is important not only for understanding the activation of signaling pathways important for T cell function but also for the development of drug targets and computer-based molecular models. In this study, phospho-specific Abs directed toward individual sites on signaling proteins were used to investigate the early phosphorylation kinetics of proteins involved in proximal TCR-induced pathways. These studies indicate that linker for activation of T cells' tyrosines have substantially different phosphorylation kinetics and that Src homology 2 domain-containing leukocyte protein of 76 kDa has rapid, transient phosphorylation kinetics compared to other proteins. In additions, we provide evidence that ZAP-70 is the primary in vivo kinase for LAT tyrosine 191 and that Itk plays a role in the phosphorylation of tyrosine 783 on phospholipase C-gamma 1. In total, these studies give new insight into the sequence, kinetics and specificity of early TCR-mediated signaling events that are vital for T cell activation.