4.7 Article

Pulsed Azidohomoalanine Labeling in Mammals (PALM) Detects Changes in Liver-Specific LKB1 Knockout Mice

Journal

JOURNAL OF PROTEOME RESEARCH
Volume 14, Issue 11, Pages 4815-4822

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.5b00653

Keywords

azidohomoalanine; newly synthesized proteins; proteomics; mass spectrometry; BONCAT; LKB1; quantitation

Funding

  1. National Institutes of Health [P41 GM103533, R01 MH067880, 1 R01 MH100175, HHSN268201000035C, R01DK080425]
  2. National Key Basic Research and Development Program of China (973) [2012CB910602]
  3. George E. Hewitt Foundation for Medical Research

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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 non-canonical 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.

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