Enzyme-catalyzed synthesis of radiolabeled 5-pregnen-3 beta-ol-20-one sulfate.
The enzymatic synthesis of [3H]5-pregnen-3 beta-ol-20-one sulfate using [3H]5-pregnen-3 beta-ol-20-one, 3'-phosphoadenosine-5'-phosphosulfate and hydroxysteroid sulfotransferase 1 purified from rat liver is reported. The described procedure allowed the obtainment of high specific activity [3H]5-pregnen-3 beta-ol-20-one sulfate in yields ranging from 78 to 86% with respect to [3H]5-pregnen-3 beta-ol-20-one. Two-dimensional thin-layer chromatography was used to purify [3H]5-pregnen-3 beta-ol-20-one sulfate which upon solvolysis resulted in the formation of [3H]5-pregnen-3 beta-ol-20-one. The identity both of the synthesized compound and the solvolysed one was confirmed by reversed-phase high pressure liquid chromatography, and 2-dimensional thin-layer chromatography.
Soliman
H R
HR
Laboratorie de Biologie Homonale, Centre Hospitalo-Universitaire St-Louis, Paris, France.
Boudou
P
P
Villette
J M
JM
Fiet
J
J
D000818:Animals; D007553:Isotope Labeling; D008099:Liver / Q000201:enzymology*; D011284:Pregnenolone / Q000737:chemistry*; D051381:Rats; D015238:Sulfotransferases / Q000378:metabolism*; D014316:Tritium
C018370:pregnenolone sulfate; D014316:Tritium; D011284:Pregnenolone; D015238:Sulfotransferases; C028152:alcohol sulfotransferase