Signaling networks that employ phosphorylation to modulate protein activities have been shown to be critically involved in the onset of diseases such as cancer. The abnormal activation of protein kinases is often either a driver or is directly resulting in the onset of these diseases. The predominance of phosphorylation as a regulator of cellular metabolism has encouraged the development of peptide arrays of kinase substrates that allow the measuring of these phosphorylation events. However, where the bioinformatics framework for DNA expression array analysis is well-developed, no advanced analysis tools are yet available for specific kinase activity profiling.
Our paper “Improved intra-array and interarray normalization of peptide microarray phosphorylation for phosphorylome and kinome profiling by rational selection of relevant spots” as published recently in Nature Scientific Reports describes a novel interarray normalization procedure, named Repetitive Signal Enhancement, or RSE, which provides a mathematical approach to limit false negatives typically generated by traditional normalization procedures. The in silico and biological experiments shown in this paper demonstrate that RSE yields superior results as compared to classical analysis tools for kinome profiling, resulting in better insights in cellular physiology.
Based on this novel approach Pepscope has developed a workflow measuring the activity of hundreds of human kinases simultaneously on a peptide array, coupled to advanced analysis tools providing unsurpassed kinase mapping in cells and tissues.