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Identification of a Tumor Specific, Active-Site Mutation in Casein Kinase 1α by Chemical Proteomics.

PLoS ONE. 2016; 
OkerbergEric S,HainleyAnna,BrownHeidi,AbanArwin,AlemayehuSenait,ShihAnn,WuJane,PatricelliMatthew P,KozarichJohn W,NomanbhoyTyzoon,RosenblumJonath
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Mutagenesis Services … plasmid encoding DDK-tagged human CSNK1A1 was obtained from Origene (catalogue number RC217936, corresponding to NCBI reference sequence NP_001020276.1, splice form 1). Mutagenesis of Asp 136 to Asn (GAC—>AAC) was performed at GenScript USA, Inc Get A Quote

摘要

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 ... More

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