Fine-tuning of substrate preferences of the Src-family kinase Lck revealed through a high-throughput specificity screen
- University of California, Berkeley, United States
- University of California, San Francisco, United States
Abstract
The specificity of tyrosine kinases is predominantly attributed 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.
Data availability
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PhosphoSitePlus Kinase-substrate datasethttps://www.phosphosite.org/staticDownloads.action.
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UniProt: the universal protein knowledgebasehttps://www.uniprot.org/help/about.
Article and author information
Author details
Funding
Damon Runyon Cancer Research Foundation
- Neel H Shah
National Institutes of Health (P01 AI091580)
- Arthur Weiss
German Academic Exchange Service London
- Mark Löbel
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2018, Shah et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Fine-tuning of substrate preferences of the Src-family kinase Lck revealed through a high-throughput specificity screen
eLife 7:e35190.
https://doi.org/10.7554/eLife.35190
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