PT  - JOURNAL ARTICLE
AU  - Neel H. Shah
AU  - Mark Löbel
AU  - Arthur Weiss
AU  - John Kuriyan
TI  - Fine-tuning of substrate preferences of the Src-family kinase Lck revealed through a high-throughput specificity screen
AID  - 10.1101/250753
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 250753
4099  - http://biorxiv.org/content/early/2018/01/19/250753.short
4100  - http://biorxiv.org/content/early/2018/01/19/250753.full
AB  - To obtain a comprehensive map of the intrinsic specificities of tyrosine kinase domains, we developed a high-throughput method that uses bacterial surface-display and next-generation sequencing to analyze the specificity of any tyrosine kinase against a library of thousands of peptides derived from human tyrosine phosphorylation sites. Using this approach, we identified a difference in the electrostatic recognition of substrates between the cytoplasmic Src-family tyrosine kinases Lck and c-Src. This divergence likely reflects the specialization of Lck to act in concert with the tyrosine kinase ZAP-70 in T cell receptor signaling. The current understanding of substrate recognition by tyrosine kinases emphasizes the role of localization by non-catalytic domains, but our results point to the importance of direct recognition at the kinase active site in fine-tuning specificity. Our method provides a simple approach that leverages next-generation sequencing to readily map the specificity of any tyrosine kinase at the proteome level.