Skip to main content

Carbonylation of adipose proteins in obesity and insulin resistance: identification of adipocyte fatty acid-binding protein as a cellular target of 4-hydroxynonenal.

Publication ,  Journal Article
Grimsrud, PA; Picklo, MJ; Griffin, TJ; Bernlohr, DA
Published in: Mol Cell Proteomics
April 2007

Obesity is a state of mild inflammation correlated with increased oxidative stress. In general, pro-oxidative conditions lead to production of reactive aldehydes such as trans-4-hydroxy-2-nonenal (4-HNE) and trans-4-oxo-2-nonenal implicated in the development of a variety of metabolic diseases. To investigate protein modification by 4-HNE as a consequence of obesity and its potential relationship to the development of insulin resistance, proteomics technologies were utilized to identify aldehyde-modified proteins in adipose tissue. Adipose proteins from lean insulin-sensitive and obese insulin-resistant C57Bl/6J mice were incubated with biotin hydrazide and detected using horseradish peroxidase-conjugated streptavidin. High carbohydrate, high fat feeding of mice resulted in a approximately 2-3-fold increase in total adipose protein carbonylation. Consistent with an increase in oxidative stress in obesity, the abundance of glutathione S-transferase A4 (GSTA4), a key enzyme responsible for metabolizing 4-HNE, was decreased approximately 3-4-fold in adipose tissue of obese mice. To identify specific carbonylated proteins, biotin hydrazide-modified adipose proteins from obese mice were captured using avidin-Sepharose affinity chromatography, proteolytically digested, and subjected to LC-ESI MS/MS. Interestingly enzymes involved in cellular stress response, lipotoxicity, and insulin signaling such as glutathione S-transferase M1, peroxiredoxin 1, glutathione peroxidase 1, eukaryotic elongation factor 1alpha-1 (eEF1alpha1), and filamin A were identified. The adipocyte fatty acid-binding protein, a protein implicated in the regulation of insulin resistance, was found to be carbonylated in vivo with 4-HNE. In vitro modification of adipocyte fatty acid-binding protein with 4-HNE was mapped to Cys-117, occurred equivalently using either the R or S enantiomer of 4-HNE, and reduced the affinity of the protein for fatty acids approximately 10-fold. These results indicate that obesity is accompanied by an increase in the carbonylation of a number of adipose-regulatory proteins that may serve as a mechanistic link between increased oxidative stress and the development of insulin resistance.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Mol Cell Proteomics

DOI

ISSN

1535-9476

Publication Date

April 2007

Volume

6

Issue

4

Start / End Page

624 / 637

Location

United States

Related Subject Headings

  • Tandem Mass Spectrometry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Proteomics
  • Protein Carbonylation
  • Obesity
  • Mice, Inbred C57BL
  • Mice
  • Insulin Resistance
  • Glutathione Transferase
  • Fatty Acid-Binding Proteins
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Grimsrud, P. A., Picklo, M. J., Griffin, T. J., & Bernlohr, D. A. (2007). Carbonylation of adipose proteins in obesity and insulin resistance: identification of adipocyte fatty acid-binding protein as a cellular target of 4-hydroxynonenal. Mol Cell Proteomics, 6(4), 624–637. https://doi.org/10.1074/mcp.M600120-MCP200
Grimsrud, Paul A., Matthew J. Picklo, Timothy J. Griffin, and David A. Bernlohr. “Carbonylation of adipose proteins in obesity and insulin resistance: identification of adipocyte fatty acid-binding protein as a cellular target of 4-hydroxynonenal.Mol Cell Proteomics 6, no. 4 (April 2007): 624–37. https://doi.org/10.1074/mcp.M600120-MCP200.
Grimsrud, Paul A., et al. “Carbonylation of adipose proteins in obesity and insulin resistance: identification of adipocyte fatty acid-binding protein as a cellular target of 4-hydroxynonenal.Mol Cell Proteomics, vol. 6, no. 4, Apr. 2007, pp. 624–37. Pubmed, doi:10.1074/mcp.M600120-MCP200.

Published In

Mol Cell Proteomics

DOI

ISSN

1535-9476

Publication Date

April 2007

Volume

6

Issue

4

Start / End Page

624 / 637

Location

United States

Related Subject Headings

  • Tandem Mass Spectrometry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Proteomics
  • Protein Carbonylation
  • Obesity
  • Mice, Inbred C57BL
  • Mice
  • Insulin Resistance
  • Glutathione Transferase
  • Fatty Acid-Binding Proteins