Source 1primary paper2025MED
Toolkit/ProKAS module
ProKAS module
Also known as: MKS module, Multiplexed Kinase Sensor module
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
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.
Usefulness & Problems
Why this is useful
The ProKAS module is an engineered tandem array of peptide sensors that enables simultaneous monitoring of multiple kinase activities. In the abstract, the reported implementation tracks ATR, ATM, and CHK1 responses to genotoxic drugs.; simultaneous monitoring of multiple kinase activities; multiplexed spatial and kinetic kinase sensing
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The ProKAS module is an engineered tandem array of peptide sensors that enables simultaneous monitoring of multiple kinase activities. In the abstract, the reported implementation tracks ATR, ATM, and CHK1 responses to genotoxic drugs.
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simultaneous monitoring of multiple kinase activities
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multiplexed spatial and kinetic kinase sensing
Problem solved
It solves the need to monitor several DNA damage response kinases at once rather than one at a time. The module also supports spatial comparison across subcellular compartments.; enables simultaneous monitoring of ATR, ATM, and CHK1 activities
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It solves the need to monitor several DNA damage response kinases at once rather than one at a time. The module also supports spatial comparison across subcellular compartments.
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enables simultaneous monitoring of ATR, ATM, and CHK1 activities
Problem links
enables simultaneous monitoring of ATR, ATM, and CHK1 activities
LiteratureIt solves the need to monitor several DNA damage response kinases at once rather than one at a time. The module also supports spatial comparison across subcellular compartments.
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It solves the need to monitor several DNA damage response kinases at once rather than one at a time. The module also supports spatial comparison across subcellular compartments.
Published Workflows
Objective: Develop a mass-spectrometry-based proteomic sensor system for quantitative, multiplexed, and spatial monitoring of kinase signaling in cells, including DNA damage response kinases and expandable design to other kinases.
Why it works: The workflow combines multiplexed peptide sensors with amino acid barcodes and mass spectrometry so multiple kinase-responsive substrates can be measured together, including across subcellular contexts. The in silico design component is intended to generate specific substrate peptides for expansion to other kinases.
monitoring kinase activity through substrate peptide phosphorylation readoutsresolving compartment-specific kinase signaling differencesmass spectrometrytandem peptide sensor arraysamino acid barcodingin silico substrate peptide design
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Target processes
recombinationselectionsignalingInput: Chemical
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The module depends on peptide sensors with amino acid barcodes and mass-spectrometry-based analysis. Its use in cells under genotoxic perturbation is explicitly described.; requires tandem peptide sensor arrays; requires amino acid barcoding
Independent follow-up evidence is still limited. Validation breadth across biological contexts is still narrow. Independent reuse still looks limited, so the evidence base may be fragile.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successMammalian Cell Lineapplication demo
mass spectrometry
Inferred from claim cl3 during normalization. An engineered ProKAS module simultaneously monitored ATR, ATM, and CHK1 activities in response to genotoxic drugs and revealed differences across nucleus, cytosol, and replication factories. Derived from claim cl3. Quoted text: 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.
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Supporting Sources
Ranked Claims
An engineered ProKAS module simultaneously monitored ATR, ATM, and CHK1 activities in response to genotoxic drugs and revealed differences across nucleus, cytosol, and replication factories.
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.
ProKAS uses a tandem array of peptide sensors with amino acid barcodes to enable multiplexed spatial, kinetic, and screening analyses.
ProKAS is based on a tandem array of peptide sensors with amino acid barcodes that allow multiplexed analysis for spatial, kinetic, and screening applications.
ProKAS is a versatile system for systematic and spatial probing of kinase action in cells.
Overall, ProKAS is a versatile system for systematically and spatially probing kinase action in cells.
ProKAS is a proteomic kinase activity sensor technique for analysis of kinase signaling using mass spectrometry.
Here we develop a proteomic kinase activity sensor technique (ProKAS) for the analysis of kinase signaling using mass spectrometry.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug prokas-module
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.
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application scopesupports
An engineered ProKAS module simultaneously monitored ATR, ATM, and CHK1 activities in response to genotoxic drugs and revealed differences across nucleus, cytosol, and replication factories.
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.
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method capabilitysupports
ProKAS uses a tandem array of peptide sensors with amino acid barcodes to enable multiplexed spatial, kinetic, and screening analyses.
ProKAS is based on a tandem array of peptide sensors with amino acid barcodes that allow multiplexed analysis for spatial, kinetic, and screening applications.
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Comparisons
Source-backed strengths
supports simultaneous monitoring of multiple kinases; uses amino acid barcodes for multiplexing
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supports simultaneous monitoring of multiple kinases
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uses amino acid barcodes for multiplexing
Compared with CfRhPDE1
ProKAS module and CfRhPDE1 address a similar problem space because they share recombination, selection, signaling.
Shared frame: same top-level item type; shared target processes: recombination, selection, signaling
Strengths here: looks easier to implement in practice.
Compared with Product Nkabinde
ProKAS module and Product Nkabinde address a similar problem space because they share selection, signaling.
Shared frame: same top-level item type; shared target processes: selection, signaling; same primary input modality: chemical
Compared with ProKAS
ProKAS module and ProKAS address a similar problem space because they share recombination, selection, signaling.
Shared frame: shared target processes: recombination, selection, signaling; same primary input modality: chemical
Ranked Citations
- 1.