Isolation of rodent airway epithelial cell proteins facilitates in vivo proteomics studies of lung toxicity

Åsa M. Wheelock, Lu Zhang, Mai Uyen Tran, Dexter Morin, Sharron Penn, Alan R Buckpitt, Charles G. Ploppet

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Recent developments in genomics, proteomics, and metabolomics hold substantial promise for understanding cellular responses to toxicants. Gene expression profiling is now considered standard procedure, but numerous publications reporting a lack of correlation between mRNA and protein expression emphasize the importance of conducting parallel proteomics studies. The cellular complexity of the lung presents great challenges for in vivo proteomics, and improved isolation methods for proteins from specific lung cell phenotypes are required. To address this issue, we have developed a novel method for isolation of rodent airway epithelial cell proteins that facilitates in vivo proteomics studies of two target-cell phenotypes of the lung, Clara cells and ciliated cells. The airway epithelial cell proteins are reproducibly solubilized, leaving the underlying basement membrane and smooth muscle intact as shown by histopathological analyses. The method yields epithelial cell-specific proteins in fivefold higher concentrations and reduces the yield of nonepithelial cell proteins 13-fold compared with homogenates from microdissected airways. In addition, 36% more protein spots were detectable by two-dimensional gel electrophoresis.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume286
Issue number2 30-2
StatePublished - Feb 2004

Keywords

  • Actin
  • Airway epithelium
  • Cytochrome P-450 monooxygenase
  • Proteomics
  • Two-dimensional gel electrophoresis

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

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