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Updated: Jan 19, 2026

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay

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A Photoaffinity Displacement Assay and Probes to Study the Cyclin-Dependent Kinase Family

Emma K Grant1,2, David J Fallon1,2, H Christian Eberl3

  • 1GlaxoSmithKline, Gunnels Wood Road, Stevenage, Hertfordshire, SG1 2NY, UK.

Angewandte Chemie (International Ed. in English)
|September 14, 2019
PubMed

Insights

Researchers developed novel photoaffinity probes to target cyclin-dependent kinases (CDKs), crucial for cell cycle control and cancer therapy. This new tool aids in profiling CDK inhibitors, potentially minimizing toxicity and advancing cancer treatment strategies.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Chemical Biology

Background:

  • Cyclin-dependent kinases (CDKs) are vital regulators of the cell cycle.
  • CDK dysregulation and mutations are linked to cancer development.
  • Targeting CDKs offers a promising therapeutic strategy for cancer, but isoform-selective inhibition is key to reduce toxicity.

Purpose of the Study:

  • To design and characterize photoaffinity probes for the CDK family.
  • To investigate the crosslinking site and enrichment capabilities of the optimized probe.
  • To develop a photoaffinity probe-based competition assay for profiling CDK inhibitors.

Main Methods:

  • Design and synthesis of photoaffinity probes targeting CDKs.
  • Investigation of probe crosslinking site and enrichment efficiency in cell lysates.
  • Development of a competition assay using photoaffinity probes to profile inhibitor binding.

Main Results:

  • Successful design of photoaffinity probes capable of binding to the CDK family.
  • Characterization of the optimized probe's crosslinking site and its ability to enrich CDKs.
  • Proof-of-concept demonstration of a competition assay for CDK inhibitor profiling.

Conclusions:

  • Photoaffinity probes offer a valuable tool for studying CDK family interactions.
  • The developed assay enables efficient profiling of CDK inhibitors.
  • This approach has broad applicability for investigating small molecule binding to protein families.
Keywords:
cancercyclin-dependent kinasesinhibitorsphotoaffinity labelingproteomics

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