Illuminating the dark kinome: utilizing multiplex peptide activity arrays to functionally annotate understudied...

Area of Science:

• Biochemistry
• Molecular Biology
• Neuroscience

Background:

• Protein kinases regulate essential biological processes but many remain understudied.
• Targeted research has focused on a subset of kinases, leaving a significant portion of the kinome unexplored.
• Understanding understudied kinases is crucial for advancing biomedical research and disease treatment.

Purpose of the Study:

• To extend the functional knowledge of understudied kinases using a hybrid empirical and in-silico approach.
• To investigate the roles of five dark tyrosine kinases (AATK, EPHA6, INSRR, LTK, TNK1) in schizophrenia, Alzheimer’s dementia (AD), and major depressive disorder (MDD).
• To identify novel kinase-substrate interactions and associated functional networks.

Main Methods:

• Utilized multiplex peptide activity arrays for empirical analysis.
• Employed robust in-silico analyses to extend functional knowledge.
• Confirmed putative substrates using biochemical assays and mass spectrometry.

Main Results:

• Identified 195 novel kinase-substrate interactions with varying affinities.
• Linked extended functional networks of understudied kinases to biological processes implicated in psychiatric and neurological disorders.
• Confirmed a putative substrate for EPHA6, a dark tyrosine kinase, and examined its network in schizophrenia.

Conclusions:

• The hybrid approach successfully extended functional knowledge of understudied kinases.
• Identified kinase-substrate interactions and networks provide actionable insights for drug development.
• This work advances the understanding of signaling networks in neurological and psychiatric diseases.