Publications

• Lecithin:retinol acyltransferase in retinal pigment epithelial microsomes.

  Saari J.C., Bredberg D.L.

  Microsomal preparations from retinal pigment epithelium carry out phosphatidylcholine synthesis upon incubation with 1-palmitoyllysophosphatidylcholine and fatty acyl-CoA. Phosphatidylcholine synthesized in situ in this manner is an acyl donor for retinyl ester synthesis, demonstrating the existen ... >> More

  Microsomal preparations from retinal pigment epithelium carry out phosphatidylcholine synthesis upon incubation with 1-palmitoyllysophosphatidylcholine and fatty acyl-CoA. Phosphatidylcholine synthesized in situ in this manner is an acyl donor for retinyl ester synthesis, demonstrating the existence of lecithin:retinol acyltransferase. Although acyl transfer to retinol is from the 1-position of phosphatidylcholine, the fatty acid in the 2-position is important in substrate recognition. The finding of this novel enzyme activity in retinal pigment epithelial microsomes suggests that phosphatidylcholine is the endogenous acyl donor in CoA-independent retinol esterification observed in these preparations. << Less

  J. Biol. Chem. 264:8636-8640(1989) [PubMed] [EuropePMC]

• Retinol esterification in bovine retinal pigment epithelium: reversibility of lecithin:retinol acyltransferase.

  Saari J.C., Bredberg D.L., Farrell D.F.

  Esterification of all-trans-retinol is a key reaction of the vertebrate visual cycle, since it produces an insoluble, relatively non-toxic, form of the vitamin for storage and supplies substrate for the isomerization reaction. CoA-dependent and -independent pathways have been described for retinol ... >> More

  Esterification of all-trans-retinol is a key reaction of the vertebrate visual cycle, since it produces an insoluble, relatively non-toxic, form of the vitamin for storage and supplies substrate for the isomerization reaction. CoA-dependent and -independent pathways have been described for retinol esterification in retinal pigment epithelium (RPE). The CoA-independent reaction, catalysed by lecithin:retinol acyltransferase (LRAT) was examined in more detail in this study. Addition of retinol to RPE microsomes results in a burst of retinyl ester synthesis, followed by a rapid apparent cessation of the reaction. However, [3H]retinol, added when retinyl ester synthesis has apparently ceased, is rapidly incorporated into retinyl ester without a net increase in the amount of ester. The specific radioactivities of [3H]retinol and [3H]retinyl ester reach the same value. [14C]Palmitate from palmitoyl-CoA is incorporated into preexisting retinyl ester in the absence of net ester synthesis, too. These exchange reactions suggest that the reaction has reached equilibrium at the plateau of the progress curve and that only the accumulation of retinyl ester, and not its synthesis, has stopped during this phase of the reaction. Studies with geometrical isomers of retinol revealed that the rate of exchange of all-trans-retinol with all-trans-retinyl esters was about 6 times more rapid than exchange of 11-cis-retinol with 11-cis-retinyl ester. This is the first demonstration of the reversibility of LRAT and the first example of stereospecificity of retinyl ester synthesis in the visual system. Reversal of the LRAT reaction could contribute to the mobilization of 11-cis-retinol from 11-cis-retinyl ester pools. << Less

  Biochem J 291:697-700(1993) [PubMed] [EuropePMC]

• Distribution of 11-cis LRAT, 11-cis RD and 11-cis REH in bovine retinal pigment epithelium membranes.

  Mata N.L., Tsin A.T.

  Our recent finding of the co-localization of 11-cis retinyl esters and 11-cis retinyl ester hydrolase (11-cis REH) activity in bovine retinal pigment epithelium (RPE) plasma membrane (PM) has led us to explore the possibility that the PM may provide 11-cis retinal for rhodopsin regeneration. In th ... >> More

  Our recent finding of the co-localization of 11-cis retinyl esters and 11-cis retinyl ester hydrolase (11-cis REH) activity in bovine retinal pigment epithelium (RPE) plasma membrane (PM) has led us to explore the possibility that the PM may provide 11-cis retinal for rhodopsin regeneration. In the RPE, visual chromophore is synthesized via a membrane associated 11-cis retinol dehydrogenase (11-cis RD). Accordingly, bovine RPE membranes enriched with either endoplasmic reticulum (ER) or plasma membrane (PM) enzyme markers were prepared and assayed for visual cycle enzyme activities. Pronounced 11-cis RD activity was associated with both ER- and PM-enriched membrane fractions. In contrast, 11-cis REH activity was mostly recovered in PM-enriched fractions while LRAT activity was found only in ER-enriched membranes. The finding that both 11-cis retinol and 11-cis retinal can be produced at the PM of the bovine RPE strongly suggests that 11-cis retinyl esters at this subcellular locale serve as a precursor of visual chromophore for pigment regeneration. << Less

  Biochim Biophys Acta 1394:16-22(1998) [PubMed] [EuropePMC]

• Evidence for a lecithin-retinol acyltransferase activity in the rat small intestine.

  MacDonald P.N., Ong D.E.

  Cellular retinol-binding protein, type II (CRBP (II] is an abundant protein of the mature enterocytes of the small intestine. It has been shown to direct retinol to an acyl-CoA-independent esterifying activity that utilizes an endogenous acyl donor (Ong, D.E., Kakkad, B., and MacDonald, P.N. (1987 ... >> More

  Cellular retinol-binding protein, type II (CRBP (II] is an abundant protein of the mature enterocytes of the small intestine. It has been shown to direct retinol to an acyl-CoA-independent esterifying activity that utilizes an endogenous acyl donor (Ong, D.E., Kakkad, B., and MacDonald, P.N. (1987) J. Biol. Chem. 262, 2729-2736). Here we report that this activity in intestinal microsomes will catalyze the transfer of acyl moieties from exogenous phosphatidylcholine (PC) to retinol-CRBP(II) to produce retinyl esters. The microsomal activity displayed positional selectivity as only the sn-1-acyl moiety of PC was transferred to retinol-CRBP(II). The retinyl ester synthase was selective for PC substrates as acyl transfer from phosphatidylethanolamine, phosphatidic acid, or free fatty acid to retinol-CRBP(II) was not observed. Some formation of retinyl esters was observed with exogenous acyl-CoA, but the amount produced was considerably lower than ester formation from exogenous PC and could be shown to be due to a different enzyme activity. Inhibitor studies clearly distinguished between the enzyme activities responsible for the acyl-CoA-dependent esterification and the phosphatidylcholine-dependent esterification of retinol. The results provide strong evidence that retinol-CRBP(II) esterification in the intestine proceeds via a phosphatidylcholine-dependent transacylase mechanism similar to that established for the esterification of cholesterol by lecithin-cholesterol acyltransferase. << Less

  J. Biol. Chem. 263:12478-12482(1988) [PubMed] [EuropePMC]

• Molecular and biochemical characterization of lecithin retinol acyltransferase.

  Ruiz A., Winston A., Lim Y.-H., Gilbert B.A., Rando R.R., Bok D.

  The enzyme responsible for conversion of all-trans-retinol 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 ... >> More

  The enzyme responsible for conversion of all-trans-retinol 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 cDNA confirmed the ability of the transfected cells to convert [3H]all-trans-retinol into authentic [3H]all-trans-retinyl palmitate as chemically determined. << Less

  J. Biol. Chem. 274:3834-3841(1999) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.