Retinol esterification in Sertoli cells by lecithin-retinol acyltransferase.
Esterification of retinol occurs during the metabolism of vitamin A in the testis. An acyl-CoA:retinol acyltransferase (ARAT) activity has been described for microsomes isolated from testis homogenates. That activity was also observed here in microsomal preparations obtained from cultured Sertoli cells from 20-day-old (midpubertal) rats. ARAT catalyzed the synthesis of retinyl laurate when free retinol and lauroyl-CoA were provided as substrates. However, in the absence of exogenous acyl-CoA, retinol was esterified by a different activity in a manner similar to the lecithin:retinol acyltransferase (LRAT) activity described recently for liver and intestine. Microsomal preparations obtained from enriched Sertoli cell fractions from the adult rat testis had 75-fold higher levels of LRAT than the preparations from midpubertal animals, but ARAT activity was the same in both these preparations. LRAT utilized an endogenous acyl donor and either unbound retinol or retinol complexed with cellular retinol-binding protein (CRBP) to catalyze the synthesis of retinyl linoleate, retinyl oleate, retinyl palmitate, and retinyl stearate. The addition of exogenous dilaurylphosphatidylcholine (DLPC) resulted in the synthesis of retinyl laurate. The esterification from both exogenous DLPC and endogenous acyl donor was inhibited by 2 mM phenylmethanesulfonyl fluoride (PMSF). ARAT activity was not affected by similar concentrations of PMSF. Furthermore, retinol bound to CRBP, a protein known to be present in Sertoli cells, was not an effective substrate for testicular ARAT. When retinol uptake and metabolism were examined in cultured Sertoli cells from 20-day-old rats, the cells synthesized the same retinyl esters that were produced by microsomal LRAT in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
Shingleton
J L
JL
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
Skinner
M K
MK
Ong
D E
DE
D000217:Acyltransferases / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002478:Cells, Cultured; D004951:Esterification; D008055:Lipids / Q000302:isolation & purification; D008297:Male; D008861:Microsomes; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D051381:Rats; D012708:Sertoli Cells / Q000187:drug effects / Q000378:metabolism*; D013737:Testis; D014801:Vitamin A / Q000378:metabolism*
D008055:Lipids; D014801:Vitamin A; D010664:Phenylmethylsulfonyl Fluoride; D000217:Acyltransferases; C056942:lecithin-retinol acyltransferase