Discovery and validation of colonic tumor-associated proteins via metabolic labeling and stable isotopic dilution

Edward L. Huttlin, Xiaodi Chen, Gregory A. Barrett-Wilt, Adrian D. Hegeman, Richard B. Halberg, Amy C. Harms, Michael A. Newton, William F. Dove, Michael R. Sussman

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

The unique biology of a neoplasm is reflected by its distinct molecular profile compared with normal tissue. To understand tumor development better, we have undertaken a quantitative proteomic search for abnormally expressed proteins in colonic tumors from ApcMin/+ (Min) mice. By raising pairs of Min and wild-type mice on diets derived from natural-abundance or 15N-labeled algae, we used metabolic labeling to compare protein levels in colonic tumor versus normal tissue. Because metabolic labeling allows internal control throughout sample preparation and analysis, technical error is minimized as compared with in vitro labeling. Several proteins displayed altered expression, and a subset was validated via stable isotopic dilution using synthetic peptide standards. We also compared gene and protein expression among tumor and nontumor tissue, revealing limited correlation. This divergence was especially pronounced for species showing biological change, highlighting the complementary perspectives provided by transcriptomics and proteomics. Our work demonstrates the power of metabolic labeling combined with stable isotopic dilution as an integrated strategy for the identification and validation of differentially expressed proteins using rodent models of human disease.

Original languageEnglish (US)
Pages (from-to)17235-17240
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume106
Issue number40
DOIs
StatePublished - Oct 6 2009

Keywords

  • Colon cancer
  • Differential mass spectrometry
  • Min mouse
  • Proteomics
  • Transcriptomics

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