Fatty acyl CoA-dependent and -independent retinol esterification by rat liver and lactating mammary gland microsomes.
Retinol esterification was examined in microsomes from rat liver and lactating mammary gland as a function of the form of retinol substrate, dependence on fatty acyl CoA, and sensitivity to phenylmethylsulfonyl fluoride (PMSF). Retinol bound to cellular retinol-binding protein (CRBP) or dispersed in solvent was esterified in a fatty acyl CoA-independent, PMSF-sensitive reaction, consistent with lecithin:retinol acyltransferase (LRAT) activity. LRAT activity exhibited the same Km (2 microM retinol) between tissues but a higher Vmax in liver as compared to that in mammary gland (47 vs 8 pmol/min/mg microsome protein, respectively). Solvent-dispersed retinol was also esterified in a fatty acyl CoA-dependent, PMSF-resistant reaction, consistent with acyl CoA:retinol acyltransferase (ARAT) activity. Retinol bound to CRBP was not a good substrate for this reaction. ARAT activity displayed a similar Vmax (300 pmol/min/mg microsome protein) between tissues but Km values of 15 and 5 microM for retinol and fatty acyl CoA in mammary gland as compared to 30 and 25 microM, respectively, in the liver. Thus, when substrate was near or below Km, retinol esterification occurred predominantly by LRAT in the liver and ARAT in the mammary gland, respectively. The concentration of CRBP in the cytosol, determined by Western blotting, was approximately 2 microM in the liver but was almost nondetectable in the mammary gland. These data suggest that retinol esterification is regulated via different mechanisms in liver and mammary gland and support a specific role for CRBP in the liver.
Randolph
R K
RK
Department of Physiology and Biochemistry, Medical College of Pennsylvania, Philadelphia 19129.
Winkler
K E
KE
Ross
A C
AC
D000214:Acyl Coenzyme A / Q000378:metabolism*; D000818:Animals; D003600:Cytosol / Q000378:metabolism; D004952:Esters; D005260:Female; D007700:Kinetics; D007774:Lactation / Q000502:physiology*; D008099:Liver / Q000378:metabolism*; D008297:Male; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D010171:Palmitoyl Coenzyme A / Q000378:metabolism; D051381:Rats; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains; D012177:Retinol-Binding Proteins / Q000378:metabolism; D054840:Retinol-Binding Proteins, Cellular; D014801:Vitamin A / Q000378:metabolism*
D000214:Acyl Coenzyme A; D004952:Esters; D012177:Retinol-Binding Proteins; D054840:Retinol-Binding Proteins, Cellular; D014801:Vitamin A; C017585:oleoyl-coenzyme A; D010171:Palmitoyl Coenzyme A