| First Author | Zhao J | Year | 2014 |
| Journal | Invest Ophthalmol Vis Sci | Volume | 55 |
| Issue | 12 | Pages | 8241-50 |
| PubMed ID | 25414195 | Mgi Jnum | J:230293 |
| Mgi Id | MGI:5755929 | Doi | 10.1167/iovs.14-15709 |
| Citation | Zhao J, et al. (2014) Preparative and biosynthetic insights into pdA2E and isopdA2E, retinal-derived fluorophores of retinal pigment epithelial lipofuscin. Invest Ophthalmol Vis Sci 55(12):8241-50 |
| abstractText | PURPOSE: Retinal-derived fluorophores that accumulate as RPE lipofuscin are implicated in pathological mechanisms of AMD. One component of RPE lipofuscin has been characterized as pdA2E, a pyridinium adduct derived from all-trans-retinal and excess ethanolamine. One-step preparation and biosynthetic studies of pdA2E and its novel isomer called isopdA2E are reported. METHODS: Biosynthetic reaction mixtures, RPE/choroids and neural retinas dissected from bovines, eyes harvested from Abca4(-/-)Rdh8(-/-) mice, irradiated samples, and enzyme-treated solutions were analyzed by HPLC, mass spectrometry, nuclear magnetic resonance spectroscopy, fluorescence spectrophotometry, and density functional theory (DFT). RESULTS: Optimization of the in vitro synthesis of pdA2E resulted in a biomimetic preparation of this pigment in a yield of 15%; this protocol also allowed the identification of isopdA2E, a double-bond isomer of pdA2E at the C13C14 position in bovine RPE lipofuscin. Interconversion between these two molecules occurs when either pdA2E or isopdA2E is exposed to light. A phospholipase D-based assay demonstrated the possibility of pdA2-PE being formed in neural retina and served as a precursor of pdA2E in the biosynthetic pathway. DFT calculations revealed that the 492-nm absorbance was assigned to the long arm of pdA2E/isopdA2E and the 340/342-nm absorbance to the short arm. Fluorescence efficiency of pdA2E and isopdA2E is very similar, but is much weaker in comparison with A2E, isoA2E, and iisoA2E. CONCLUSIONS: Our results facilitate the understanding of compositions and biosynthetic pathways of adverse RPE lipofuscin. |
