ABSTRACT
The [3H]corticosterone-transcortin complexes from kidney cytosol show elution positions on DEAE-cellulose identical to serum transcortin. The incorporation of 14C-labeled amino acids into anti-transcortin-precipitable material of kidney slices has been measured and compared with that of serum transcortin. It was established that kidney synthesized transcortin with an apparent molecular weight of 66 kDa on SDS-electrophoresis which resembles serum corticosteroid-binding globulin. Studies on the binding of [125I]anti-transcortin-IgG to membrane-bound rat kidney polyribosomes revealed an association of [125I]anti-transcortin-IgG with a discrete polyribosome fraction in the heavy polyribosome region; free polyribosomes were devoid of antigenic material able to bind antibodies to transcortin.
Subject(s)
Kidney/metabolism , Polyribosomes/metabolism , Transcortin/metabolism , Animals , Cell Compartmentation , Corticosterone/metabolism , Cytosol/metabolism , Immunologic Techniques , Molecular Weight , Rats , Transcortin/biosynthesisABSTRACT
Transcortin biosynthesis in rat has been examined using liver slices technique. The incorporation of 14C-labeled amino acids into the anti-transcortin-precipitable material of liver slices has been measured and compared with that of serum transcortin. It was shown that liver synthesized transcortin with an apparent mol. wt of 66 kDa on SDS-electrophoresis which co-migrated with authentic rat serum transcortin. In order to determine an intracellular distribution of transcortin synthesizing polyribosomes, the binding character of [125J]anti-transcortin-IgG to free and membrane-bound rat liver polyribosomes has been studied. It was shown that after incubation of [125J]anti-transcortin-IgG with liver membrane-bound polyribosomes, the radioactivity was associated with the discrete polyribosome fraction in heavy polyribosome region. In similar experiments the radioactivity of [125J]anti-transcortin-IgG bound to the free polyribosomes was distributed throughout the polyribosome region. The results of our experiments obviously demonstrated that serum transcortin was synthesized exclusively on membrane-bound polyribosomes of rat liver; free polyribosomes were devoid of detectable antigenic material able to bind antibodies to transcortin.
