The identification of tyrosine phosphorylation-dependent interactome of immune coreceptors is crucial for the understanding of signal pathways involved in immunotherapy. However, identifying motif-specific interactome for each tyrosine phosphorylation site commonly found on these multi-phosphorylated membrane proteins remains challenging. Here we describe a photoaffinity-based chemical proteomic approach with synthetic full-length CD28 cytoplasmic tails (CD28cyto) to dissect the motif-specific cytoplasmic interactome of the critical immune coreceptor CD28 by covalent capturing and label-free quantification. Our chemical proteomic analysis well recapitulated reported CD28cyto interacting proteins to pY191 and pY209 motifs. We defined the stand-alone interaction of phospholipase PLCG1 to Y191 motif with enhanced affinity to the sequence neighboring the transmembrane domain. Importantly, we explored the interactome of previously undefined pY218 motif and verified that a critical kinase PKC? strongly and directly associated with pY218 through its C2 domain. This synthetic CD28cyto-based photoaffinity proteomic approach is generically applicable to study other immune coreceptors with multiple pY sites on their linear cytoplasmic tails.
[doi:10.25345/C5MV2G]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: CD28-cyto interactome
Principal Investigators: (in alphabetical order) |
Ruijun Tian, SUSTech, China |
Submitting User: | Mercutio |
Xiong Chen, Shanping Ji, Zheyi Liu, Xiao Yuan, Congsheng Xu, Ruxi Qi, An He, Heng Zhao, Haiping Song, Chunlei Xiao, Weina Gao, Peng R. Chen, Ray Luo, Pengfei Li, Fangjun Wang, Xueming Yang, and Ruijun Tian.
Motif-dependent Immune Coreceptor Interactome Profiling by Photoaffinity Chemical Proteomics.
Cell Chemical Biology (2022), https://doi.org/10.1016/j.chembiol.2022.01.005.
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