Characterization of dihydro-A2PE: an intermediate in the A2E biosynthetic pathway.
Bisretinoid lipofuscin pigments that accumulate in retinal pigment epithelial cells are implicated in the etiology of several forms of macular degeneration, including juvenile onset Stargardt disease, Best vitelliform macular degeneration, and age-related macular degeneration. One of these compounds, A2E, is generated by phosphate hydrolysis of a phosphatidyl-pyridinium bisretinoid (A2PE) that forms within photoreceptor outer segments. Here, we demonstrate that the formation of the aromatic pyridinium ring of A2PE follows from the oxidation of a dihydropyridinium intermediate. Time-dependent density functional theory calculation, based on the structure of dihydro-A2E, produced a simulated UV-visible absorbance spectrum characterized by maxima of 494 and 344 nm. Subsequently, a compound exhibiting similar UV-visible absorbance maxima (lambdamax 490 and 330 nm) was identified in the A2E biomimetic reaction mixture. By liquid chromatography-mass spectrometry (LC-MS) this bischromophore had the expected mass of the dihydro-pyridinium bisretinoid. The compound also exhibited the behavior of a biosynthetic intermediate since it formed in advance of the final product A2E and was consumed as A2E accumulated. Moreover, under deoxygenated conditions, conversion to the aromatic pyridinium bisretinoid was inhibited. Taken together, these findings indicate that A2E biosynthesis involves the oxidation of a dihydropyridinium intermediate dihydro-A2PE. An understanding of the biosynthetic pathways of retinal pigment epithelial lipofuscin pigments is critical to the development of therapies for macular degeneration that are based on limiting the formation of these damaging compounds.
Kim
So R
SR
Department of Ophthalmology, Columbia University, New York, New York 10032, USA.
He
Jiangtao
J
Yanase
Emiko
E
Jang
Young P
YP
Berova
Nina
N
Sparrow
Janet R
JR
Nakanishi
Koji
K
D002851:Chromatography, High Pressure Liquid; D003198:Computer Simulation; D004847:Epithelial Cells; D008062:Lipofuscin / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism*; D008268:Macular Degeneration / Q000378:metabolism; D013058:Mass Spectrometry; D008954:Models, Biological; D010084:Oxidation-Reduction; D011726:Pyridinium Compounds / Q000737:chemistry / Q000378:metabolism*; D012168:Retinal Pigments / Q000096:biosynthesis*; D012176:Retinoids / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D014801:Vitamin A / Q000737:chemistry / Q000378:metabolism*
C112040:A2-E (N-retinylidene-N-retinylethanolamine); D008062:Lipofuscin; D011726:Pyridinium Compounds; D012168:Retinal Pigments; D012176:Retinoids; D014801:Vitamin A