Toward quantitative phosphotyrosine profiling in vivo.

TitleToward quantitative phosphotyrosine profiling in vivo.
Publication TypeJournal Article
Year of Publication2012
AuthorsJohnson, H, White, FM
JournalSemin Cell Dev Biol
Date Published2012 Oct
KeywordsAnimals, Humans, Phosphotyrosine, Proteomics, Reproducibility of Results, Signal Transduction, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Tyrosine phosphorylation is a dynamic reversible post-translational modification that regulates many aspects of cell biology. To understand how this modification controls biological function, it is necessary to not only identify the specific sites of phosphorylation, but also to quantify how phosphorylation levels on these sites may be altered under specific physiological conditions. Due to its sensitivity and accuracy, mass spectrometry (MS) has widely been applied to the identification and characterization of phosphotyrosine signaling across biological systems. In this review we highlight the advances in both MS and phosphotyrosine enrichment methods that have been developed to enable the identification of low level tyrosine phosphorylation events. Computational and manual approaches to ensure confident identification of phosphopeptide sequence and determination of phosphorylation site localization are discussed along with methods that have been applied to the relative quantification of large numbers of phosphorylation sites. Finally, we provide an overview of the challenges ahead as we extend these technologies to the characterization of tyrosine phosphorylation signaling in vivo. With these latest developments in analytical and computational techniques, it is now possible to derive biological insight from quantitative MS-based analysis of signaling networks in vitro and in vivo. Application of these approaches to a wide variety of biological systems will define how signal transduction regulates cellular physiology in health and disease.

Alternate JournalSemin. Cell Dev. Biol.
PubMed ID22677333
PubMed Central IDPMC3496061
Grant ListR01 CA118705 / CA / NCI NIH HHS / United States
U54 CA112967 / CA / NCI NIH HHS / United States