15N-labeled brain enables quantification of proteome and phosphoproteome in cultured primary neurons.

ABSTRACT: Terminally differentiated primary cells represent a valuable in vitro model to study signaling events associated within a specific tissue. Quantitative proteomic methods using metabolic labeling in primary cells encounter labeling efficiency issues hindering the use of these cells. Here we developed a method to quantify the proteome and phosphoproteome of cultured neurons using (15)N-labeled brain tissue as an internal standard and applied this method to determine how an inhibitor of an excitatory neural transmitter receptor, phencyclidine (PCP), affects the global phosphoproteome of cortical neurons. We identified over 10,000 phosphopeptides and made accurate quantitative measurements of the neuronal phosphoproteome after neuronal inhibition. We show that short PCP treatments lead to chan

SUBMITTER: Liao L 

PROVIDER: S-EPMC3279198 | biostudies-literature | 2012 Feb

REPOSITORIES: biostudies-literature