Structural characterization of bisretinoid A2E photocleavage products and implications for age-related macular degeneration.
Fluorescent bisretinoids, such as A2E and all-trans-retinal dimer, form as a by-product of vitamin A cycling in retina and accumulate in retinal pigment epithelial (RPE) cells as lipofuscin pigments. These pigments are implicated in pathological mechanisms involved in several vision-threatening diseases including age-related macular degeneration. Efforts to understand damaging events initiated by these bisretinoids have revealed that photoexcitation of A2E by wavelengths in the visible spectrum leads to singlet oxygen production and photooxidation of A2E. Here we have employed liquid chromatography coupled to electrospray ionization mass spectrometry together with tandem mass spectrometry (MS/MS), to demonstrate that A2E also undergoes photooxidation-induced degradation and we have elucidated the structures of some of the aldehyde-bearing cleavage products. Studies in which A2E was incubated with a singlet oxygen generator yielded results consistent with a mechanism involving bisretinoid photocleavage at sites of singlet molecular oxygen addition. We provide evidence that one of the products released by A2E photodegradation is methylglyoxal, a low molecular weight reactive dicarbonyl with the capacity to form advanced glycation end products. Methylglyoxal is already known to be generated by carbohydrate and lipid oxidation; this is the first report of its production via bisretinoid photocleavage. It is significant that AGE-modified proteins are detected in deposits (drusen) that accumulate below RPE cells in vivo; drusen have been linked to age-related macular degeneration pathogenesis. Whereas various processes play a role in drusen formation, these findings are indicative of a contribution from lipofuscin photooxidation in RPE.
Wu
Yalin
Y
Department of Ophthalmology, Columbia University, 630 West 168th Street, New York, NY 10032, USA.
Yanase
Emiko
E
Feng
Xidong
X
Siegel
Marshall M
MM
Sparrow
Janet R
JR
D000447:Aldehydes / Q000737:chemistry; D002241:Carbohydrates / Q000737:chemistry; D006801:Humans; D008027:Light; D008055:Lipids / Q000737:chemistry; D008062:Lipofuscin / Q000737:chemistry; D008268:Macular Degeneration / Q000628:therapy*; D010100:Oxygen / Q000737:chemistry; D010777:Photochemistry / Q000379:methods; D011726:Pyridinium Compounds / Q000737:chemistry*; D011765:Pyruvaldehyde / Q000737:chemistry; D055213:Retinal Pigment Epithelium / Q000166:cytology*; D012176:Retinoids / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014801:Vitamin A / Q000737:chemistry
C112040:A2-E (N-retinylidene-N-retinylethanolamine); D000447:Aldehydes; D002241:Carbohydrates; D008055:Lipids; D008062:Lipofuscin; D011726:Pyridinium Compounds; D012176:Retinoids; D014801:Vitamin A; D011765:Pyruvaldehyde; D010100:Oxygen