Proteins modified by malondialdehyde, 4-hydroxynonenal, or advanced glycation end products in lipofuscin of human retinal pigment epithelium.
OBJECTIVE
Lipofuscin (LF) accumulation in the retinal pigment epithelium (RPE) is associated with age and various retinal diseases. Toxic LF compounds may interfere with normal RPE function. Oxidative modification of proteins was determined in LF granules from human eyes.


METHODS
LF was isolated from the RPE-choroid complex of 10 pairs of donor eyes by gradient ultracentrifugation. Protein compounds were separated by two-dimensional (2-D) gel electrophoresis and screened by Western blot analysis for lipid peroxidation- or glucoxidation-induced damage-in particular, by malondialdehyde (MDA), 4-hydroxynonenal (HNE), and advanced glycation end products (AGEs). Identity of the immunostained proteins was revealed using 2-D software for comparison of the spot position with Coomassie-stained 2-D gels of the same samples.


RESULTS
By comparing the results taken from the authors' previous proteome analysis of RPE LF with an immunoblot analysis of the same samples, this study shows that a variety of LF-associated proteins were damaged by aberrant covalent modifications of MDA, 4-HNE, and AGEs. Several proteins were altered by two or three different modification types. Modified mitochondrial proteins indicated that autophagy of altered proteins also contributed to lipofuscin formation.


CONCLUSIONS
The identification of lipid peroxidation and glucoxidation products in proteinaceous LF components in human RPE supports the hypothesis that these compounds are involved in lipofuscinogenesis and may contribute to the cytotoxic effects of LF in retinal diseases such as age-related macular degeneration and Stargardt disease. Their identification may help to identify potential future treatment targets.
Schutt
Florian
F
Department of Ophthalmology, University of Heidelberg, Heidelberg, Germany. florian_schuett@med.uni-heidelberg.de
Bergmann
Marion
M
Holz
Frank G
FG
Kopitz
Jurgen
J
D000368:Aged; D000369:Aged, 80 and over; D000447:Aldehydes / Q000494:pharmacology*; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005136:Eye Proteins / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D006801:Humans; D015227:Lipid Peroxidation; D008062:Lipofuscin / Q000302:isolation & purification / Q000378:metabolism*; D008315:Malondialdehyde / Q000494:pharmacology*; D010084:Oxidation-Reduction; D010857:Pigment Epithelium of Eye / Q000187:drug effects* / Q000378:metabolism; D020543:Proteome / Q000378:metabolism
D000447:Aldehydes; D005136:Eye Proteins; D017127:Glycation End Products, Advanced; D008062:Lipofuscin; D020543:Proteome; D008315:Malondialdehyde; C027576:4-hydroxy-2-nonenal