Targeted Quantification of Protein Phosphorylation and Its Contributions towards Mathematical Modeling of Signaling Pathways

Abstract

Post-translational modifications (PTMs) are key regulatory mechanisms that can control protein function. Of these, phosphorylation is the most common and widely studied. Because of its importance in regulating cell signaling, precise and accurate measurements of protein phosphorylation across wide dynamic ranges are crucial to understanding how signaling pathways function. Although immunological assays are commonly used to detect phosphoproteins, their lack of sensitivity, specificity, and selectivity often make them unreliable for quantitative measurements of complex biological samples. Recent advances in Mass Spectrometry (MS)-based targeted proteomics have made it a more useful approach than immunoassays for studying the dynamics of protein phosphorylation. Selected reaction monitoring (SRM)—also known as multiple reaction monitoring (MRM)—and parallel reaction monitoring (PRM) can quantify relative and absolute abundances of protein phosphorylation in multiplexed fashions targeting specific pathways. In addition, the refinement of these tools by enrichment and fractionation strategies has improved measurement of phosphorylation of low-abundance proteins. The quantitative data generated are particularly useful for building and parameterizing mathematical models of complex phospho-signaling pathways. Potentially, these models can provide a framework for linking analytical measurements of clinical samples to better diagnosis and treatment of disease.

Low‐intensity pulsed ultrasound induces proliferation of human neural stem cells

“…Moreover, as multiple signalling pathways are elicited by LIPUS such as PI3K‐Akt and ERK/MAPK, 51 ERK, JNK or JNK and p38, 52 ERK, JNK and p38 MAPK, 70 PI3K‐Akt, 59 notch, 15 GSK‐3β/β‐catenin, 13 and PI3K‐Akt and JNK MAPK, 55 Ca 2 + signalling, 11 further investigations are required to reveal all involved signalling pathways induced by LIPUS and their interplays in the neural stem cells. In this context, advanced techniques such as proteomic quantification 71,72 or genomic techniques 73,74 can be used to reveal all involved pathways, their regulators and their interplays.…”