TY - JOUR TI - Fine-tuning of substrate preferences of the Src-family kinase Lck revealed through a high-throughput specificity screen AU - Shah, Neel H AU - Löbel, Mark AU - Weiss, Arthur AU - Kuriyan, John A2 - Cole, Philip A. VL - 7 PY - 2018 DA - 2018/03/16 SP - e35190 C1 - eLife 2018;7:e35190 DO - 10.7554/eLife.35190 UR - https://doi.org/10.7554/eLife.35190 AB - The specificity of tyrosine kinases is attributed predominantly to localization effects dictated by non-catalytic domains. We developed a method to profile the specificities of tyrosine kinases by combining bacterial surface-display of peptide libraries with next-generation sequencing. Using this, we showed that the tyrosine kinase ZAP-70, which is critical for T cell signaling, discriminates substrates through an electrostatic selection mechanism encoded within its catalytic domain (Shah et al., 2016). Here, we expand this high-throughput platform to analyze the intrinsic specificity of any tyrosine kinase domain against thousands of peptides derived from human tyrosine phosphorylation sites. Using this approach, we find a difference in the electrostatic recognition of substrates between the closely related Src-family kinases Lck and c-Src. This divergence likely reflects the specialization of Lck to act in concert with ZAP-70 in T cell signaling. These results point to the importance of direct recognition at the kinase active site in fine-tuning specificity. KW - tyrosine kinase KW - T cell KW - substrate specificity JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -