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Published on: February 21, 2019
Identification of a Tumor Specific, Active-Site Mutation in Casein Kinase 1α by Chemical Proteomics
Eric S Okerberg1, Anna Hainley1, Heidi Brown1
1ActivX Biosciences, Inc., La Jolla, CA, United States of America.
Abstract:
We describe the identification of a novel, tumor-specific missense mutation in the active site of casein kinase 1α (CSNK1A1) using activity-based proteomics. Matched normal and tumor colon samples were analyzed using an ATP acyl phosphate probe in a kinase-targeted LC-MS2 platform. An anomaly in the active-site peptide from CSNK1A1 was observed in a tumor sample that was consistent with an altered catalytic aspartic acid. Expression and analysis of the suspected mutant verified the presence of asparagine in the probe-labeled, active-site peptide for CSNK1A1. Genomic sequencing of the colon tumor samples confirmed the presence of a missense mutation in the catalytic aspartic acid of CSNK1A1 (GAC→AAC). To our knowledge, the D163N mutation in CSNK1A1 is a newly defined mutation to the conserved, catalytic aspartic acid of a protein kinase and the first missense mutation identified using activity-based proteomics. The tumorigenic potential of this mutation remains to be determined.
Insights
Researchers identified a new tumor-specific mutation in casein kinase 1 alpha (CSNK1A1) using activity-based proteomics. This novel D163N mutation in the kinase
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Casein kinase 1 alpha (CSNK1A1) is a key regulator of cellular processes.
- Mutations in protein kinases can drive tumorigenesis.
- Activity-based proteomics enables the discovery of enzyme activity and modifications.
Purpose of the Study:
- To identify novel, tumor-specific kinase mutations using activity-based proteomics.
- To characterize a newly identified mutation in the CSNK1A1 active site.
Main Methods:
- Activity-based proteomics utilizing an ATP acyl phosphate probe.
- Kinase-targeted liquid chromatography-tandem mass spectrometry (LC-MS2).
- Analysis of matched normal and colon tumor samples.
- Expression and genomic sequencing for mutation verification.
Main Results:
- A novel, tumor-specific missense mutation (D163N) was identified in the CSNK1A1 active site.
- The mutation alters the conserved catalytic aspartic acid residue.
- This is the first missense mutation in CSNK1A1's catalytic aspartic acid and the first identified via activity-based proteomics.
Conclusions:
- A novel, tumor-specific CSNK1A1 mutation (D163N) was discovered using activity-based proteomics.
- This mutation affects a critical residue in the kinase active site.
- The tumorigenic potential of this novel mutation warrants further investigation.
