Application of photoaffinity labeling with [11,12-3H]all-trans-retinoic acid to characterization of rat liver microsomal UDP-glucuronosyltransferase(s) with activity toward retinoic acid.
[3H]All-trans-retinoic acid has been shown to be an effective photoaffinity label for microsomal UDP-glucuronosyltransferases. Labeling of rat liver microsomal proteins with [3H]all-trans-retinoic acid and [32P]5-azido-UDP-glucuronic acid has shown that at least one protein in the 50-56 kDa mass range encompassing the UDP-glucuronosyltransferases photoincorporated both probes. The fraction of solubilized microsomal protein eluted from a UDP-hexanolamine affinity column with 50 microM UDP-glucuronic acid contained two protein bands, both of which photoincorporated [3H] all-trans-retinoic acid and were detected on Western blot by anti-UDP-glucuronosyltransferase antibodies. Enzymatic glucuronidation activity toward atRA in the same fraction was enriched five-fold over that of native or solubilized microsomes.
Little
J M
JM
Division of Gastroenterology, Department of Internal Medicine, University of Arkansas for Medical Sciences, Little Rock 72205, USA.
Radominska
A
A
D000345:Affinity Labels; D000818:Animals; D002846:Chromatography, Affinity; D004789:Enzyme Activation; D005965:Glucuronates / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology* / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D051381:Rats; D017207:Rats, Sprague-Dawley; D012685:Sepharose; D014212:Tretinoin / Q000378:metabolism*; D014230:Triazoles; D014316:Tritium; D014530:Uridine Diphosphate
D000345:Affinity Labels; D005965:Glucuronates; D010761:Phosphorus Radioisotopes; D014230:Triazoles; D014316:Tritium; D014212:Tretinoin; D014530:Uridine Diphosphate; D012685:Sepharose; D014453:Glucuronosyltransferase