title: Thyrotropin alters the utilization of thyroglobulin's hormonogenic sites.
abstract: We injected rabbits and guinea pigs with bovine thyrotropin (TSH) daily for 3 days, while controls received saline. All animals received sodium [125I]iodide on the second day, and thyroglobulin was purified from the thyroids of each group by gel filtration. Hormonogenic tryptic peptides from each S-cyanoethylated thyroglobulin preparation were isolated by high performance liquid chromatography, and their amino acid sequences were determined, permitting their localization within the thyroglobulin polypeptide chain by comparison with cDNA-derived sequences from bovine and human thyroglobulins. Thyroglobulins from the saline-injected rabbits and guinea pigs contained the same four major hormonogenic sites, designated A-D, previously described (Dunn, J. T., Anderson, P. C., Fox, J. W., Fassler, C. A., Dunn, A. D., Hite, L. A., and Moore, R. C. (1987) J. Biol. Chem. 262, 16948-16952). In both species, sites A and C were the major loci for thyroxine and triiodothyronine, respectively. However, site D in the guinea pig had a greater ratio of [125I]thyroxine to [127I]thyroxine than did site A, whereas the reverse was true in the rabbit. TSH administration produced the following changes in thyroglobulins of both species, relative to controls: 1) an increase in the ratio of [125I]triiodothyronine to [125I] thyroxine (rabbit, 0.29 versus 0.17; guinea pig, 0.19 versus 0.08), with the increase in triiodothyronine principally at site C; 2) a marked increase in 125I/127I and in thyroxine formation at site D (14.1% of thyroglobulin's thyroxine versus 9.8% in rabbits, 24 versus 13% in guinea pigs); 3) a corresponding decrease in thyroxine formation at site A (33 versus 43% in rabbits, 30 versus 46% in guinea pigs); and 4) a sharp increase in conversion of thyroglobulin's N-terminal 125I-labeled approximately 20 kDa hormone-rich iodopeptide, which contains site A, to a 125I-labeled approximately 15-kDa (rabbit) or 125I-labeled approximately 13-kDa (guinea pig) form, reflecting probable peptide bond cleavage. Our results show that TSH alters both the structure of the thyroglobulin molecule and the priority of utilization of its hormonogenic sites. We conclude that these changes are important to TSH's enhancement of thyroid hormone synthesis.
authors.affiliation: Department of Internal Medicine, University of Virginia School of Medicine, Charlottesville 22908.
authors.forename: C A
authors.initials: CA
authors.lastname: Fassler
authors.forename: J T
authors.initials: JT
authors.lastname: Dunn
authors.forename: P C
authors.initials: PC
authors.lastname: Anderson
authors.forename: J W
authors.initials: JW
authors.lastname: Fox
authors.forename: A D
authors.initials: AD
authors.lastname: Dunn
authors.forename: L A
authors.initials: LA
authors.lastname: Hite
authors.forename: R C
authors.initials: RC
authors.lastname: Moore
authors.forename: P S
authors.initials: PS
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D005260:Female; D006168:Guinea Pigs; D007454:Iodides / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D011817:Rabbits; D012016:Reference Values; D013045:Species Specificity; D013954:Thyroglobulin / Q000235:genetics / Q000378:metabolism*; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism*; D013963:Thyroid Hormones / Q000096:biosynthesis; D013972:Thyrotropin / Q000494:pharmacology*; D014357:Trypsin
chemical_list: D007454:Iodides; D007457:Iodine Radioisotopes; D010446:Peptide Fragments; D013963:Thyroid Hormones; D013972:Thyrotropin; D013954:Thyroglobulin; D014357:Trypsin