Isolation and one-step preparation of A2E and iso-A2E, fluorophores from human retinal pigment epithelium.
Age-related macular degeneration, a major cause of blindness for which no satisfactory treatments exist, leads to a gradual decrease in central high acuity vision. The accumulation of fluorescent materials, called lipofuscin, in retinal pigment epithelial cells of the aging retina is most pronounced in the macula. One of the fluorophores of retinal pigment epithelial lipofuscin has been characterized as A2E, a pyridinium bis-retinoid, which is derived from two molecules of vitamin A aldehyde and one molecule of ethanolamine. An investigation aimed at optimizing the in vitro synthesis of A2E has resulted in the one-step biomimetic preparation of this pigment in 49% yield, readily producing more than 50 mg in one step. These results have allowed for the optimization of HPLC conditions so that nanogram quantities of A2E can be detected from extracts of tissue samples. By using 5% of the extract from individual aged human eyes, this protocol has led to the quantification of A2E and the characterization of iso-A2E, a new A2E double bond isomer; all-trans-retinol and 13-cis-retinol also have been identified in these HPLC chromatograms. Exposure of either A2E or iso-A2E to light gives rise to 4:1 A2E:iso-A2E equilibrium mixtures, similar to the composition of these two pigments in eye extracts. A2E and iso-A2E may exhibit surfactant properties arising from their unique wedge-shaped structures.
Parish
C A
CA
Department of Chemistry, Columbia University, New York, NY 10027, USA.
Hashimoto
M
M
Nakanishi
K
K
Dillon
J
J
Sparrow
J
J
D006801:Humans; D010857:Pigment Epithelium of Eye / Q000737:chemistry*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification; D012168:Retinal Pigments / Q000737:chemistry / Q000302:isolation & purification*; D012176:Retinoids / Q000737:chemistry / Q000302:isolation & purification*
C112040:A2-E (N-retinylidene-N-retinylethanolamine); D020033:Protein Isoforms; D012168:Retinal Pigments; D012176:Retinoids