Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells.

Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Research Abstract Details 

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Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Abstract Text:

Chandima Abeywickrama,Hiroko Matsuda,Steffen Jockusch,Jilin Zhou,Young P Jang,Bi-Xing Chen,Yasuhiro Itagaki,Bernard F Erlanger,Koji Nakanishi,Nicholas J Turro,Janet R Sparrow,

The autofluorescent lipofuscin pigment A2E accumulates in retinal pigment epithelial cells with age and is particularly abundant in some retinal disorders. To generate a polyclonal antibody that recognizes this pyridinium bisretinoid molecule, we immunized rabbits with bovine serum albumin (BSA) conjugates in which the protein was linked to the A2E molecule via its pyridinium ethanolamine moiety. Analysis by matrix-assisted laser desorption ionization/time of flight mass spectrometry (MALDI-TOF MS) of the A2E-BSA conjugate indicated the presence of five intact A2E molecules covalently linked to BSA, thus deeming it a suitable antigen for immunization. By immunocytochemical staining, the rabbit polyclonal antibody recognized A2E that had accumulated in cultured cells, whereas dot-blot analysis revealed binding to both A2E and A2E-rabbit serum albumin (A2E-RSA) conjugate but no cross-reactivity with various retinoids. Preimmune serum was nonreactive. In fluorescence spectroscopy studies, antibody-A2E binding was evidenced by a fluorescence increase and by a blue-shift in the emission maximum consistent with a change in A2E milieu upon antibody binding. The changes in fluorescence emission upon antibody binding could reflect several processes including restrictions on trans-cis isomerization and intersystem crossing of photo-excited A2E.

Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Publishing Authors By Initials

C Abeywickrama,H Matsuda,S Jockusch,J Zhou,YP Jang,BX Chen,Y Itagaki,BF Erlanger,K Nakanishi,NJ Turro,JR Sparrow,

For similar investigative techniques: chemistry, analytical: photometry: spectrophotometry: spectrophotometry, ultraviolet research abstracts see: investigative techniques: chemistry, analytical: photometry: spectrophotometry: spectrophotometry, ultraviolet research

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Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Proceedings of the National Academy of Sciences of

VOLUME: 104

Page Numbers: 14610-5

Journal Abbreviation: Proc. Natl. Acad. Sci. U.S.A.

ISSN: 0027-8424

DAY: 5

MONTH: 09

YEAR: 2007

Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 7505876

Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Keywords Mesh Terms:

KEYWORDS: Spectrophotometry, Ultraviolet

MESH TERMS: metabolism

Chemical & Substance for Abstract: Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells. Information

Substance Name: Serum Albumin, Bovine

Registry Number: 0

Grant and Affiliation Information for Immunochemical recognition of A2E, a pigment in the lipofuscin of retinal pigment epithelial cells.

AFFILIATION: Department of Chemistry, Columbia University, New York, NY 10027, USA.

Country: United States

AGENCY: United States NHLBI

GRANT: HL 4737

ACRONYM: HL

MEDLINETA: Proc Natl Acad Sci U S A

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