Palmitoyl transferase activity of lecithin retinol acyl transferase.
Lecithin retinol acyl transferase (LRAT) has the essential role of catalyzing the transfer of an acyl group from the sn-1 position of lecithin to vitamin A to generate all-trans-retinyl esters (tREs). In vitro studies had shown previously that LRAT also can exchange palmitoyl groups between RPE65, a tRE binding protein essential for vision, and tREs. This exchange is likely to be of regulatory significance in the operation of the visual cycle. In the current study, the substrate specificity of LRAT is explored with palmitoylated amino acids and dipeptides as RPE65 surrogates. Both O- and S-substituted palmitoylated analogues are excellent substrates for tLRAT, a readily expressed and readily purified form of LRAT. Using vitamin A as the palmitoyl acceptor, tREs are readily formed. The cognate of these reactions occurs in crude retinal pigment epithelial (RPE) membranes as well. RPE membranes containing LRAT transfer palmitoyl groups from radiolabeled [1-(14)C]-l-alpha-dipalmitoyl diphosphatidylcholine (DPPC) to RPE65. Palmitoyl transfer is abolished by preincubation with a specific LRAT antagonist both in membranes and with purified tLRAT. These experiments are consistent with an expanded role for LRAT function as a protein palmitoyl transferase.
Xue
Linlong
L
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 45 Shattuck Street, Boston, Massachusetts 02115, USA.
Jahng
Wan Jin
WJ
Gollapalli
Deviprasad
D
Rando
Robert R
RR
D000217:Acyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D002417:Cattle; D005136:Eye Proteins / Q000737:chemistry*; D066298:In Vitro Techniques; D015394:Molecular Structure; D010170:Palmitoyl-CoA Hydrolase; D010455:Peptides / Q000737:chemistry; D010857:Pigment Epithelium of Eye / Q000201:enzymology*; D013379:Substrate Specificity
D005136:Eye Proteins; D010455:Peptides; D000217:Acyltransferases; C056942:lecithin-retinol acyltransferase; D010170:Palmitoyl-CoA Hydrolase