Mitogen activated protein kinases (MAPKs) regulate a series of cellular functions, and abnormal changes in the activity of MAPKs can be associated with many diseases. Since different MAPK isoforms have special contributions to certain cellular malfunctions, the testing methods of their individual isoform activity are important. Instead of using highly selective biosensors in combination with inhibitors for off-target kinases for one single isoform, our group reported differential sensing techniques with chemometric tools to detect several isoforms. Our original SOX-peptide library shows great quantitative ability for extracellular signal-regulated kinases (ERK) in the presence of other two major subfamilies. In this work, we developed an expanded library with elevated cross-reactivity, intentionally included more peptides with affinity to Jun-N-terminal kinases (JNK) and p38 stress activated kinases (p38). The obtained fingerprints of kinase identity and activity were depicted using 3D linear discriminant analysis (LDA) plot, showing excellent classification of all the isoforms. (Fig.1). Next, we performed a quantitative assay for each subfamily in the presence of others, then built the calibration models by a support vector machine (SVM) regression method. In future work we’ll use complex training sets, aimed at detecting ERK/JNK/p38 activity simultaneously in a single assay.

Differential sensing of multiplex MAPK isoforms using expanded SOX-peptide library

2019 Fall National Meeting

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