# Proteomic Sensors for Quantitative, Multiplexed and Spatial Monitoring of Kinase Signaling

## TL;DR

This paper introduces a new method to measure and track kinase activity in cells using proteomic sensors, enabling detailed spatial and quantitative analysis.

## Contribution

The novel ProKAS platform allows multiplexed, spatial, and quantitative kinase activity monitoring using mass spectrometry.

## Key findings

- ProKAS can simultaneously monitor ATR, ATM, and CHK1 kinase activities in response to DNA damage.
- The platform reveals spatial differences in kinase signaling within the nucleus, cytosol, and replication factories.
- An in silico method was developed to design 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 developed a proteomic kinase activity sensor platform (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 novel versatile system for systematically and spatially probing kinase action in cells.