Retinal proteins modified by 4-hydroxynonenal: Identification of molecular targets

Becky Kapphahn, Babatomiwa M. Giwa, Kristin M. Berg, Heidi Roehrich, Xiao Feng, Timothy W. Olsen, Deborah A Ferrington

Research output: Contribution to journalArticle

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Abstract

The reactive aldehyde, 4-hydroxynonenal (HNE), is a product of lipid peroxidation that can covalently modify and inactivate proteins. Previously, we reported increased HNE modification of select retinal proteins resolved by one-dimensional gel electrophoresis in aged Fisher 344 × Brown Norway rats (Louie, J.L., Kapphahn, R.J., Ferrington, D.A., 2002. Proteasome function and protein oxidation in the aged retina. Exp. Eye Res. 75, 271-284). In the current study, quantitative assessment of HNE molar content using slot blot immunoassays showed HNE content is increased 30% in aged rat retina. In contrast, there was no age-related difference in HNE content in individual spots resolved by 2D gel electrophoresis suggesting the increased modification is likely on membrane proteins that are missing on 2D gels. The HNE-immunoreactive proteins resolved by 2D gel electrophoresis were identified by MALDI-TOF mass spectrometry. These proteins are involved in metabolism, chaperone function, and fatty acid transport. Proteins that were frequently modified and had the highest molar content of HNE included triosephosphate isomerase, α enolase, heat shock cognate 70 and βB2 crystallin. Immunochemical detection of HNE adducts on retinal sections showed greater immune reaction in ganglion cells, photoreceptor inner segment, and the inner plexiform layer. Identification of HNE modified proteins in two alternative model systems, human retinal pigment epithelial cells in culture (ARPE19) and human donor eyes, indicated that triosephosphate isomerase and α enolase are generally modified. These results identify a common subset of proteins that contain HNE adducts and suggest that select retinal proteins are molecular targets for HNE modification.

Original languageEnglish (US)
Pages (from-to)165-175
Number of pages11
JournalExperimental Eye Research
Volume83
Issue number1
DOIs
StatePublished - Jul 1 2006

Fingerprint

Proteins
Triose-Phosphate Isomerase
Phosphopyruvate Hydratase
Electrophoresis, Gel, Two-Dimensional
Retina
Gels
4-hydroxy-2-nonenal
Photoreceptor Cells
Crystallins
Retinal Pigments
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Proteasome Endopeptidase Complex
Immunoassay
Aldehydes
Ganglia
Lipid Peroxidation
Electrophoresis
Shock
Mass Spectrometry
Membrane Proteins

Keywords

  • 4-hydroxynonenal
  • mass spectrometry
  • oxidation
  • proteomics
  • retina
  • two-dimensional gel electrophoresis

Cite this

Retinal proteins modified by 4-hydroxynonenal : Identification of molecular targets. / Kapphahn, Becky; Giwa, Babatomiwa M.; Berg, Kristin M.; Roehrich, Heidi; Feng, Xiao; Olsen, Timothy W.; Ferrington, Deborah A.

In: Experimental Eye Research, Vol. 83, No. 1, 01.07.2006, p. 165-175.

Research output: Contribution to journalArticle

Kapphahn, Becky ; Giwa, Babatomiwa M. ; Berg, Kristin M. ; Roehrich, Heidi ; Feng, Xiao ; Olsen, Timothy W. ; Ferrington, Deborah A. / Retinal proteins modified by 4-hydroxynonenal : Identification of molecular targets. In: Experimental Eye Research. 2006 ; Vol. 83, No. 1. pp. 165-175.
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