Version 2 2015-12-17, 10:34Version 2 2015-12-17, 10:34
Version 1 2015-11-11, 12:00Version 1 2015-11-11, 12:00
journal contribution
posted on 2015-12-17, 10:34authored byDaniel
B. McClatchy, Yuanhui Ma, Chao Liu, Benjamin D. Stein, Salvador Martínez-Bartolomé, Debbie Vasquez, Kristina Hellberg, Reuben J. Shaw, John R. Yates
Quantification
of proteomes by mass spectrometry has proven to
be useful to study human pathology recapitulated in cellular or animal
models of disease. Enriching and quantifying newly synthesized proteins
(NSPs) at set time points by mass spectrometry has the potential to
identify important early regulatory or expression changes associated
with disease states or perturbations. NSP can be enriched from proteomes
by employing pulsed introduction of the noncanonical amino acid, azidohomoalanine
(AHA). We demonstrate that pulsed introduction of AHA in the feed
of mice can label and identify NSP from multiple tissues. Furthermore,
we quantitate differences in new protein expression resulting from
CRE-LOX initiated knockout of LKB1 in mouse livers. Overall, the PALM
strategy allows for the first time in vivo labeling of mouse tissues
to differentiate protein synthesis rates at discrete time points.