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Pulsed azidohomoalanine labeling in mammals (PALM) detects changes in liver-specific LKB1 knockout mice

Academic Article
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Overview

related to degree

  • Stein, Benjamin, Ph.D. in Chemistry, Scripps Research 2008 - 2015

authors

  • McClatchy, D. B.
  • Ma, Y.
  • Liu, C.
  • Stein, Benjamin
  • Martinez-Bartolome, S.
  • Vasquez, D.
  • Hellberg, K.
  • Shaw, R. J.
  • Yates III, John

publication date

  • November 2015

journal

  • Journal of Proteome Research  Journal

abstract

  • 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.

subject areas

  • Alanine
  • Alkynes
  • Animals
  • Azides
  • Biotin
  • Click Chemistry
  • Food, Formulated
  • Gene Expression
  • Integrases
  • Liver
  • Male
  • Methionine
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Sequence Annotation
  • Protein Serine-Threonine Kinases
  • Proteome
  • Proteomics
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Research

keywords

  • BONCAT
  • LKB1
  • azidohomoalanine
  • mass spectrometry
  • newly synthesized proteins
  • proteomics
  • quantitation
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Identity

PubMed Central ID

  • PMC4642245

International Standard Serial Number (ISSN)

  • 1535-3893

Digital Object Identifier (DOI)

  • 10.1021/acs.jproteome.5b00653

PubMed ID

  • 26445171
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Additional Document Info

start page

  • 4815

end page

  • 4822

volume

  • 14

issue

  • 11

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