DeKinomics pulse-chases kinase functions in living cells

Area of Science:

• Cellular biology
• Proteomics
• Optogenetics

Background:

• Kinase function is complex and context-dependent, requiring precise tools for study.
• Understanding kinase roles in health and disease necessitates methods for rapid stimulation and proteome analysis.

Purpose of the Study:

• To develop a novel optogenetic strategy for studying kinase functions in real-time.
• To enable proteome-wide profiling of kinase-driven phosphorylation with high temporal resolution.

Main Methods:

• Developed decaging kinase coupled proteomics (DeKinomics), an optogenetic 'pulse-chase' approach.
• Utilized rapid kinase stimulation and global proteome characterization in living cells.
• Applied DeKinomics for identifying direct kinase substrates and mapping phosphorylation events.

Main Results:

• Achieved second-timescale proteome-wide profiling of kinase-driven phosphorylation.
• Successfully identified direct kinase substrates using the 'gain-of-function' feature.
• Characterized global phosphorylation of understudied receptor tyrosine kinases in native cellular contexts.

Conclusions:

• DeKinomics provides a powerful, generalizable strategy for studying kinase functions.
• The method offers high specificity and temporal resolution under living conditions.
• Enables in-depth analysis of kinase-mediated cellular processes in health and disease.