Molecular and biochemical characterization of lecithin retinol acyltransferase

Alberto Ruiz, Anette Winston, Young Hee Lim, Bryant A. Gilbert, Robert R. Rando, Dean Bok

Research output: Contribution to journalArticle

193 Citations (Scopus)

Abstract

The enzyme responsible for conversion of all-transretinol into retinyl esters, the lecithin retinol acyltransferase (LRAT) has been characterized at the molecular level. The cDNA coding for this protein was cloned and its amino acid sequence deduced. LRAT is composed of a polypeptide of 230 amino acid residues with a calculated mass of 25.3 kDa. Tissue distribution analysis by Northern blot showed expression of a 5.0-kilobase transcript in the human retinal pigment epithelium as well as in other tissues that are known for their high LRAT activity and vitamin A processing. Affinity labeling experiments using specific compounds with high affinity for LRAT and monospecific polyclonal antibodies raised in rabbits against two peptide sequences for LRAT confirmed the molecular mass of LRAT as a 25-kDa protein. High performance liquid chromatography analysis of the reaction product formed by HEK-293 cells transfected with LRAT eDNA confirmed the ability of the transfected cells to convert [3H]all-trans-retinol into authentic [3H]all-trans-retinyl palmitate as chemically determined.

Original languageEnglish
Pages (from-to)3834-3841
Number of pages8
JournalJournal of Biological Chemistry
Volume274
Issue number6
DOIs
Publication statusPublished - 1999 Feb 5
Externally publishedYes

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Vitamin A
Tissue
Amino Acids
Peptides
Retinal Pigments
Retinal Pigment Epithelium
HEK293 Cells
High performance liquid chromatography
Molecular mass
Tissue Distribution
lecithin-retinol acyltransferase
Reaction products
Northern Blotting
Labeling
Amino Acid Sequence
Esters
Proteins
Complementary DNA
High Pressure Liquid Chromatography
Rabbits

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular and biochemical characterization of lecithin retinol acyltransferase. / Ruiz, Alberto; Winston, Anette; Lim, Young Hee; Gilbert, Bryant A.; Rando, Robert R.; Bok, Dean.

In: Journal of Biological Chemistry, Vol. 274, No. 6, 05.02.1999, p. 3834-3841.

Research output: Contribution to journalArticle

Ruiz, Alberto ; Winston, Anette ; Lim, Young Hee ; Gilbert, Bryant A. ; Rando, Robert R. ; Bok, Dean. / Molecular and biochemical characterization of lecithin retinol acyltransferase. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 6. pp. 3834-3841.
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