Differential regulation of thyrotropin subunit apoprotein and carbohydrate biosynthesis by thyroid hormone.
The regulation of TSH apoprotein and carbohydrate biosynthesis by thyroid hormone was studied by incubating pituitaries from normal and hypothyroid (3 weeks post-thyroidectomy) rats in medium containing [14C]alanine and [3H] glucosamine. After 6 h, samples were sequentially treated with anti-TSH beta to precipitate TSH and free TSH beta, anti-LH beta to clear the sample of LH and free LH beta, then anti-LH alpha to precipitate free alpha-subunit. Total proteins were acid precipitated. All precipitates were subjected to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, which were then sliced and assayed by scintillation spectrometry. In hypothyroid pituitaries plus medium, [14C]alanine incorporation in combined and free beta-subunits was 26 times normal (P less than 0.001) and considerably greater than the 3.4-fold increase seen in total protein (P less than 0.05); combined and free alpha-subunits showed no specific increase in apoprotein synthesis. [3H]Glucosamine incorporation in combined alpha- and beta-subunits in hypothyroid samples was 13 and 21 times normal, respectively, and was greater than the 1.9-fold increase in total protein (P less than 0.05); free alpha-subunit showed no specific increase in carbohydrate synthesis. The glucosamine to alanine ratio, reflecting relative glycosylation of newly synthesized molecules, was increased in hypothyroidism for combined alpha-subunits (P less than 0.001), but not for combined beta-subunits, free alpha-subunits, or total proteins. In summary, short term hypothyroidism selectively stimulated TSH beta apoprotein synthesis and carbohydrate synthesis of combined alpha- and beta-subunits. Hypothyroidism also increased the relative glycosylation of combined alpha-subunit. Thus, thyroid hormone deficiency appears to alter the rate-limiting step in TSH assembly (i.e. beta-subunit synthesis) as well as the carbohydrate structure of TSH, which may play important roles in its biological function.
D000818:Animals; D001059:Apoproteins / Q000096:biosynthesis*; D002241:Carbohydrates / Q000096:biosynthesis*; D002250:Carbon Radioisotopes; D008297:Male; D010902:Pituitary Gland / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D013963:Thyroid Hormones / Q000502:physiology*; D013972:Thyrotropin / Q000096:biosynthesis* / Q000378:metabolism; D014316:Tritium
D001059:Apoproteins; D002241:Carbohydrates; D002250:Carbon Radioisotopes; D011506:Proteins; D013963:Thyroid Hormones; D014316:Tritium; D013972:Thyrotropin