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JW Crabb, M Miyagi, X Gu, KA West, A Marmorstein, M Kamei, KG Shadrach, ME Rayborn, RG Salomon, JG Hollyfield; Proteomic Approaches to Age-Related Macular Degeneration . Invest. Ophthalmol. Vis. Sci. 2002;43(13):860.
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Purpose: The causes of drusen formation and Bruch's membrane thickening in age-related macular degeneration (AMD) are not known. However, the progression of AMD might be slowed if these processes could be modulated. Proteomic studies of these tissues from normal and AMD donors seek clues to the biochemical pathways involved in AMD. Methods: Microdissection and laser capture methods are used to isolate drusen and Bruch's membrane. Mass spectrometry tools are used for protein identification. Oxidative protein modifications are identified using bioinformatic tools and antibodies. Immunocytochemistry provides confirmation of drusen and Bruch's membrane localization. Results: We have developed methods for isolating drusen and Bruch's membrane that provide microgram amounts of protein. Currently 29 potential drusen proteins have been identified by LC MS/MS, 8 of which were observed by others using immunocytochemistry. We have confirmed clusterin, calgranulin A and B, and psoriasin as new drusen components by immunocytochemistry. Western results suggest that docosahexenoate derived protein modifications (ie, carboxyethyl pyrrole adducts) are more abundant in AMD than normal tissues. Advanced glycation end products and abnormal protein crosslinks also appear in drusen and Bruch's membrane. Conclusions: Oxidative protein modifications may be causally involved in drusen formation and Bruch's membrane thickening. Protoemic studies to test this hypothesis are in progress.
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