# Proteomic sensors for quantitative multiplexed and spatial monitoring of kinase signaling

## TL;DR

The paper introduces ProKAS, a new method to measure and track kinase activity in cells with high precision and spatial detail.

## Contribution

ProKAS enables multiplexed and spatial monitoring of kinase activity using proteomic sensors and mass spectrometry.

## Key findings

- ProKAS can simultaneously monitor ATR, ATM, and CHK1 kinase activities in response to genotoxic drugs.
- The method reveals spatial differences in kinase signaling in the nucleus, cytosol, and replication factories.
- An in silico approach allows designing specific substrate peptides for other kinases.

## Abstract

Understanding kinase action requires precise quantitative measurements of their activity in vivo. In addition, the ability to capture spatial information of kinase activity is crucial to deconvolute complex signaling networks, interrogate multifaceted kinase actions, and assess drug effects or genetic perturbations. Here we develop a proteomic kinase activity sensor technique (ProKAS) for the analysis of kinase signaling using mass spectrometry. ProKAS is based on a tandem array of peptide sensors with amino acid barcodes that allow multiplexed analysis for spatial, kinetic, and screening applications. We engineered a ProKAS module to simultaneously monitor the activities of the DNA damage response kinases ATR, ATM, and CHK1 in response to genotoxic drugs, while also uncovering differences between these signaling responses in the nucleus, cytosol, and replication factories. Furthermore, we developed an in silico approach for the rational design of specific substrate peptides expandable to other kinases. Overall, ProKAS is a versatile system for systematically and spatially probing kinase action in cells.

Understanding kinase action requires precise quantitative and spatial measurements of their activity in vivo. Here the authors develop a proteomic kinase activity sensor technique (ProKAS) enabling multiplexed spatial, kinetic, and screening analyses of kinase activities via mass spectrometry.