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Quantitative mass spectrometry to investigate epidermal growth factor receptor phosphorylation dynamics
Author(s) -
Schuchardt Sven,
Borlak Jürgen
Publication year - 2007
Publication title -
mass spectrometry reviews
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 126
eISSN - 1098-2787
pISSN - 0277-7037
DOI - 10.1002/mas.20155
Subject(s) - chemistry , receptor tyrosine kinase , phosphorylation , epidermal growth factor , tyrosine phosphorylation , signal transduction , epidermal growth factor receptor , tyrosine kinase , microbiology and biotechnology , protein phosphorylation , receptor , transmembrane protein , tyrosine , cell surface receptor , grb2 , biochemistry , protein kinase a , biology
Identifying proteins of signaling networks has received much attention, because an array of biological processes are entirely dependent on protein cross‐talk and protein–protein interactions. Protein posttranslational modifications (PTM) add an additional layer of complexity, resulting in complex signaling networks. Of particular interest to our working group are the signaling networks of epidermal growth factor (EGF) receptor, a transmembrane receptor tyrosine kinase involved in various cellular processes, including cell proliferation, differentiation, and survival. Ligand binding to the N ‐terminal residue of the extracellular domain of EGF receptor induces conformational changes, dimerization, and (auto)‐phosphorylation of intracellular tyrosine residues. In addition, activated EGF receptor may positively affect survival pathways, and thus determines the pathways for tumor growth and progression. Notably, in many human malignancies exaggerated EGF receptor activities are commonly observed. An understanding of the mechanism that results in aberrant phosphorylation of EGF receptor tyrosine residues and derived signaling cascades is crucial for an understanding of molecular mechanisms in cancer development. Here, we summarize recent labeling methods and discuss the difficulties in quantitative MS‐based phosphorylation assays to probe for receptor tyrosine kinase (RTK) activity. We also review recent advances in sample preparation to investigate membrane‐bound RTKs, MS‐based detection of phosphopeptides, and the diligent use of different quantitative methods for protein labeling. © 2007 Wiley Periodicals, Inc., Mass Spec Rev 27:51–65, 2008