Proteome analysis of lipofuscin in human retinal pigment epithelial cells.
Excessive accumulation of lipofuscin in postmitotic retinal pigment epithelial cells is a common pathogenetic pathway in various blinding retinal diseases including age-related macular degeneration, which is now the most common cause of registerable blindness in the industrialized nations. To better understand the role of lipofuscin accumulation and to manipulate the pathogenetic mechanisms on both experimental and therapeutic levels we analyzed the proteome of isolated human ocular lipofuscin granules from human RPE cells. After homogenization and fractionation by gradient ultracentrifugation of the RPE/choroid complex from 10 pairs of human donors, protein compounds were separated by 2D gel electrophoresis and analyzed using matrix-assisted laser desorption/ionization mass spectrometry and HPLC-coupled electrospray tandem mass spectrometry. Besides a better understanding of downstream pathways, this approach may provide new targets for therapeutic interventions in a currently untreatable disease.
Schutt
F
F
Department of Ophthalmology, INF 400, D-69120 Heidelberg, Germany. florian_schuett@med.uni-heidelberg.de
Ueberle
B
B
Schnölzer
M
M
Holz
F G
FG
Kopitz
J
J
D002851:Chromatography, High Pressure Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008062:Lipofuscin / Q000302:isolation & purification* / Q000378:metabolism; D008268:Macular Degeneration / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D020543:Proteome; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D008062:Lipofuscin; D020543:Proteome