Immunochemical characterization of developmental changes in rat hepatic hydroxysteroid sulfotransferase.
A major isoenzyme of hepatic androsterone-sulfating sulfotransferase (AD-ST) was purified from adult female rats. The activity was purified 122-fold over that found in the cytosol and showed a single protein band with a subunit molecular mass of 30 kDa after sodium dodecyl sulfate polyacrylamide gel electrophoresis. The purified enzyme exhibited four isoelectric variants of subunits on denaturing isoelectrofocusing gels (pI = 5.8, 6.1, 6.7 and 7.2). Rabbit antiserum raised against the enzyme specifically detected AD-ST polypeptide in rat liver cytosol. Immunoblot analysis of liver cytosol from female and male rats at various ages showed good correlation between the levels of AD-ST activity and AD-ST polypeptide. Significant levels of AD-ST activity and polypeptide were detected in senescent male rats, though normal adult male rats have very low levels of AD-ST activity and protein. The relative content of the isoelectric variants of AD-ST were different in liver cytosol of weanling and adult females, indicating that age- and gender-related alterations of hepatic AD-ST activity are primarily determined by the levels of AD-ST polypeptide and the relative amounts of the four isoelectric variants of the enzyme.
Homma
H
H
Kyoritsu College of Pharmacy, Tokyo, Japan.
Nakagome
I
I
Kamakura
M
M
Matsui
M
M
D000375:Aging; D000818:Animals; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003600:Cytosol / Q000201:enzymology; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015151:Immunoblotting; D007527:Isoenzymes / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology* / Q000254:growth & development; D008297:Male; D008970:Molecular Weight; D011817:Rabbits / Q000276:immunology; D051381:Rats; D015238:Sulfotransferases; D013466:Sulfurtransferases / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*
D007527:Isoenzymes; D013466:Sulfurtransferases; D015238:Sulfotransferases; C028152:alcohol sulfotransferase