Analysis of the role of Nrf2 in the expression of liver proteins in mice using two-dimensional gel-based proteomics

Pharmacol Rep. 2012;64(3):680-97. doi: 10.1016/s1734-1140(12)70863-0.


Background: The transcription factor Nrf2 regulates expression of multiple cellular defence proteins through the antioxidant response element (ARE). Nrf2-deficient mice (Nrf2(-/-)) are highly susceptible to xenobiotic-mediated toxicity, but it is not known whether this reflects low basal expression or reduced inducibility of Nrf2-regulated genes in response to chemical insults.

Methods: Wild type and Nrf2(-/-) mice were fed diet supplemented with the established Nrf2 inducer butylated hydroxyanisole (BHA) [0.5% (w/w)] for 14 days. To define the range of Nrf2-regulated proteins, both basally and following exposure to BHA, a comparison of the liver proteomes of Nrf2(-/-) and wild type mice was conducted. The two-dimensional gel electrophoresis (2-DE) technique and MALDI mass spectrometry were utilized in the attempt to define Nrf2-regulated proteins.

Results: Overall, 24 proteins were identified, which were regulated either basally (3 proteins), inducibly (16 proteins), or both (5 proteins). These included several well-established Nrf2-driven gene products e.g., aldo-keto reductase and glutathione transferases. Multiple consensus ARE/ARE-like sequences were found in the Nrf2-regulated genes.

Conclusions: This study confirms the central role of Nrf2 in the induction of multiple defense proteins as well as its control in the constitutive expression of certain proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / metabolism
  • Aldehyde Reductase
  • Aldo-Keto Reductases
  • Animals
  • Butylated Hydroxyanisole / pharmacology*
  • Electrophoresis, Gel, Two-Dimensional
  • Female
  • Gene Expression Regulation
  • Glutathione Transferase / metabolism
  • Liver / metabolism*
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred ICR
  • Mice, Knockout
  • NF-E2 Transcription Factor, p45 Subunit / genetics*
  • Proteins / genetics*
  • Proteomics / methods
  • Response Elements
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods


  • NF-E2 Transcription Factor, p45 Subunit
  • Nfe2 protein, mouse
  • Proteins
  • Butylated Hydroxyanisole
  • Alcohol Oxidoreductases
  • Aldo-Keto Reductases
  • Aldehyde Reductase
  • AKR7A5 protein, mouse
  • Glutathione Transferase