This is a protocol preprint. Preprints are preliminary reports that have not undergone peer review. They should not be considered conclusive, used to inform clinical practice, or referenced by the media as validated information.
Method Article
High-Throughput Kinase Activity Mapping (HT-KAM) system: biochemical assay
Jean-Philippe Coppé
Kinome Activity Mapping
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Declarations:
Mapping the phospho-catalytic profile of kinase enzymes in cells or tissues remains a challenge. Here, we introduce a practical biochemical assay to measure the enzymatic activity of kinases using peptides as surrogate sensors to identify kinases in tumor biopsies and cell lines. The platform relies on collections of peptide probes that are derived from biological target sites of kinases, and that operate as distinct combinatorial peptide sets to simultaneously distinguish and measure the phospho-catalytic activity of many kinase enzymes. The assay is modular by design: users can adapt probe libraries and assay conditions to their needs. We named this functional proteomic platform 'High-Throughput Kinase Activity Mapping (HT-KAM) system'. The procedure described in this Protocol Exchange chapter focuses on detailing the biochemical assay, and is related to the Nature Cell Biology manuscript NCB-C36710 titled: "Mapping phospho-catalytic dependencies of therapy-resistant tumors reveals actionable vulnerabilities".
Keywords
kinase enzymes, phospho-catalytic activity screening, peptide sensors, combinatorial profiling, cell extracts, tissue biospecimen extracts, functional proteomics, signaling networks
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Posted 12 Jun, 2019
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Subject Areas
Cell biology
Biological techniques
Mathematics and computing
Chemical biology
Computational biology and bioinformatics
Biochemistry
Associated Publications
Mapping phospho-catalytic dependencies of therapy-resistant tumours reveals actionable vulnerabilities
by Jean-Philippe Coppé, Miki Mori, Bo Pan, +17
Nature Cell Biology (03 June, 2019)
Method Article
High-Throughput Kinase Activity Mapping (HT-KAM) system: biochemical assay
Jean-Philippe Coppé
Kinome Activity Mapping
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 12 Jun, 2019
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Declarations:
Mapping the phospho-catalytic profile of kinase enzymes in cells or tissues remains a challenge. Here, we introduce a practical biochemical assay to measure the enzymatic activity of kinases using peptides as surrogate sensors to identify kinases in tumor biopsies and cell lines. The platform relies on collections of peptide probes that are derived from biological target sites of kinases, and that operate as distinct combinatorial peptide sets to simultaneously distinguish and measure the phospho-catalytic activity of many kinase enzymes. The assay is modular by design: users can adapt probe libraries and assay conditions to their needs. We named this functional proteomic platform 'High-Throughput Kinase Activity Mapping (HT-KAM) system'. The procedure described in this Protocol Exchange chapter focuses on detailing the biochemical assay, and is related to the Nature Cell Biology manuscript NCB-C36710 titled: "Mapping phospho-catalytic dependencies of therapy-resistant tumors reveals actionable vulnerabilities".
Associated Publications
Mapping phospho-catalytic dependencies of therapy-resistant tumours reveals actionable vulnerabilities
by Jean-Philippe Coppé, Miki Mori, Bo Pan, +17
Nature Cell Biology (03 June, 2019)