EMBL Australia Node in Single Molecule Science, School of Medical Sciences, The University of New South Wales, Sydney 2052, Australia.
ARC Centre of Excellence in Advanced Molecular Imaging, The University of New South Wales, Sydney 2052, Australia.
Int J Mol Sci. 2020 May 15;21(10):3498. doi: 10.3390/ijms21103498.
T cell activation is initiated when ligand binding to the T cell receptor (TCR) triggers intracellular phosphorylation of the TCR-CD3 complex. However, it remains unknown how biophysical properties of TCR engagement result in biochemical phosphorylation events. Here, we constructed an optogenetic tool that induces spatial clustering of ζ-chain in a light controlled manner. We showed that spatial clustering of the ζ-chain intracellular tail alone was sufficient to initialize T cell triggering including phosphorylation of ζ-chain, Zap70, PLCγ, ERK and initiated Ca flux. In reconstituted COS-7 cells, only Lck expression was required to initiate ζ-chain phosphorylation upon ζ-chain clustering, which leads to the recruitment of tandem SH2 domain of Zap70 from cell cytosol to the newly formed ζ-chain clusters at the plasma membrane. Taken together, our data demonstrated the biophysical relevance of receptor clustering in TCR signaling.
T 细胞的激活是由配体与 T 细胞受体（TCR）结合触发 TCR-CD3 复合物的细胞内磷酸化而引发的。然而，目前尚不清楚 TCR 结合的生物物理特性如何导致生化磷酸化事件。在这里，我们构建了一种光遗传学工具，可通过光控制的方式诱导 ζ 链的空间聚类。我们表明，ζ 链细胞内尾部的空间聚类本身足以启动 T 细胞触发，包括 ζ 链、Zap70、PLCγ、ERK 的磷酸化，并启动 Ca 流。在重组的 COS-7 细胞中，只有在 ζ 链聚类时 Lck 的表达才足以启动 ζ 链磷酸化，这导致 Zap70 的串联 SH2 结构域从细胞质招募到质膜上新形成的 ζ 链簇。总之，我们的数据表明受体聚类在 TCR 信号中的生物物理相关性。
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