Analysis of renal and hippocampal type I and type II receptors by fast protein liquid chromatography.
Type I and Type II adrenal steroid receptors from rat renal and hippocampal cytosols were studied by the technique of Fast Protein Liquid Chromatography. Type I receptors were labelled with [3H]aldosterone plus excess RU26988, and Type II receptors with [3H]dexamethasone. On a Mono Q anion exchange column the molybdate-stabilized renal and hippocampal Type I receptors both eluted as single symmetrical peaks at 0.27 M NaCl, with a recovery of approximately 90% and 60-fold purification (renal) and 10-15-fold (hippocampal). Molybdate-stabilized Type II binding sites from both hippocampal and renal cytosols co-eluted with the Type I sites. On Superose gel filtration renal Type I receptor-steroid complexes consistently eluted two fractions later than hippocampal Type I complexes, suggesting that the renal complexes are smaller; Type II receptor-steroid complexes from both cytosols co-eluted, consistently one fraction behind hippocampal Type I sites. Sequential gel filtration and anion exchange chromatography achieved a 1000-fold purification of renal Type I binding sites, with an overall recovery of 10%.
Doyle
D
D
Medical Research Centre, Prince Henry's Hospital, Melbourne, Australia.
Krozowski
Z
Z
Morgan
F J
FJ
Funder
J W
JW
D000732:Androstanols / Q000378:metabolism; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003345:Corticosterone / Q000378:metabolism; D003600:Cytosol / Q000032:analysis; D003907:Dexamethasone / Q000378:metabolism; D006624:Hippocampus / Q000032:analysis*; D007668:Kidney / Q000032:analysis*; D051381:Rats; D018161:Receptors, Mineralocorticoid; D011987:Receptors, Steroid / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism
D000732:Androstanols; D018161:Receptors, Mineralocorticoid; D011987:Receptors, Steroid; C038341:RU 26988; D003907:Dexamethasone; D003345:Corticosterone