RT Journal Article
SR Electronic
T1 A procedure for the rapid separation and purification of UDP-glucuronosyltransferases from rabbit liver microsomes.
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 97
OP 101
VO 10
IS 2
A1 R H Tukey
A1 R Robinson
A1 B Holm
A1 C N Falany
A1 T R Tephly
YR 1982
UL http://dmd.aspetjournals.org/content/10/2/97.abstract
AB A technique has been developed which rapidly separates and purifies UDP-glucuronosyltransferases from liver microsomes of untreated rabbits. by use of this method, highly purified estrone and p-nitrophenol UDP-glucuronosyltransferases can be obtained in good yield in about 48 hr. Microsomes were solubilized with the nonionic detergent Emulgen 911 in low ionic strength buffers and applied to a DEAE-cellulose column equilibrated with low ionic strength buffers. UDP-glucuronosyltransferase activities were then eluted in a stepwise fashion with increasing concentration of KCl. Three fractions were studied. The first two fractions contained only estrone UDP-glucuronosyltransferase activity while a third contained p-nitrophenol UDP-glucuronosyltransferase activity. Each fraction was directly applied to a UDP-hexanolamine Sepharose-4B column, which was then washed extensively with KCl, and the transferases were eluted with UDP-glucuronic acid. A method for separating the transferases on the affinity column is presented. Testosterone and morphine could not be conjugated by any of the purified enzymes.