Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress

Hyunju Lee, Hyewon Chung, Hilal Arnouk, Folami Lamoke Powell, Richard C. Hunt, William J.M. Hrushesky, Patricia A. Wood, Sung Haeng Lee, Wan Jin Jahng

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The regeneration of the 11-cis-retinyl imine chromophore of rhodopsin during the visual cycle and mechanisms that control this process are central questions in the field of vision research. The retinal pigment epithelium (RPE)-specific protein RPE65 is centrally involved in the isomerization and hydrolysis of all-trans-retinyl esters. In this study, we investigated RPE65 cleavage and potential regulatory mechanisms under oxidative stress conditions. The D407 RPE cell cultures were exposed to H2O2 (100-1000μM). Changes in the levels of RPE65 and proteins related to apoptosis were investigated using gel electrophoresis and western blotting. Mass spectrometry was used to confirm the identity of RPE65. C57BL/6J (M450) and C3HeB/FeJ (L450) mice were used for in vivo experiments. We found that a novel 45kDa truncated fragment of the RPE65 protein, designated RPE45, appears in RPE cells upon light exposure or oxidative stress. RPE45 is generated in vitro by recombinant caspases via an ubiquitination-dependent mechanism. Collectively, our results indicate that oxidative stress during the visual cycle results in cleavage of RPE65.

Original languageEnglish (US)
Pages (from-to)104-108
Number of pages5
JournalInternational Journal of Biological Macromolecules
Volume47
Issue number2
DOIs
StatePublished - Aug 1 2010

Fingerprint

Oxidative stress
Retinal Pigments
Retinal Pigment Epithelium
Pigments
Oxidative Stress
Proteins
Imines
Rhodopsin
Ubiquitination
Cell death
Chromophores
Caspases
Isomerization
Electrophoresis
Cell culture
Mass spectrometry
Regeneration
Hydrolysis
Mass Spectrometry
Esters

Keywords

  • Light
  • Oxidative stress
  • RPE65
  • Retinal pigment epithelium
  • Visual cycle

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Economics and Econometrics
  • Energy(all)

Cite this

Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress. / Lee, Hyunju; Chung, Hyewon; Arnouk, Hilal; Powell, Folami Lamoke; Hunt, Richard C.; Hrushesky, William J.M.; Wood, Patricia A.; Lee, Sung Haeng; Jahng, Wan Jin.

In: International Journal of Biological Macromolecules, Vol. 47, No. 2, 01.08.2010, p. 104-108.

Research output: Contribution to journalArticle

Lee, Hyunju ; Chung, Hyewon ; Arnouk, Hilal ; Powell, Folami Lamoke ; Hunt, Richard C. ; Hrushesky, William J.M. ; Wood, Patricia A. ; Lee, Sung Haeng ; Jahng, Wan Jin. / Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress. In: International Journal of Biological Macromolecules. 2010 ; Vol. 47, No. 2. pp. 104-108.
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