Photocleavable peptide-oligonucleotide conjugates for protein kinase assays by MALDI-TOF MS.

ABSTRACT: Robust methods for highly parallel, quantitative analysis of cellular protein tyrosine kinase activities may provide tools critically needed to decipher oncogenic signaling, discover new targeted drugs, diagnose cancer and monitor patients. Here, we describe proof-of-principle for a novel protein kinase assay with the potential to help overcome these challenges. MALDI-TOF mass spectrometry provides an ideal tool for label-free multiplexed analysis of peptide phosphorylation, but is poorly matched to homogeneous assays and complex samples. Thus, we conjugated a common oligonucleotide tag to multiple peptide substrates, offering efficient capture from solution-phase kinase reactions by annealing to the complementary sequence tethered to PEG-passivated superparamagnetic microparticles. To ena

SUBMITTER: Zhou G 

PROVIDER: S-EPMC3462237 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature