title: Thyroid hormone residues are released from thyroglobulin with only limited alteration of the thyroglobulin structure.
abstract: We have tried to characterize thyroglobulin (Tg) degradation products in purified pig thyroid lysosomes to determine whether the release of thyroid hormone residues from Tg involves a random proteolytic attack or discrete and selective cleavage reactions. The intralysosomal soluble protein fraction was prepared by osmotic pressure-dependent lysis of lysosomes purified by isopycnic centrifugation on Percoll gradients. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate revealed the presence of a fraction of Tg (5-10% of total lysosomal protein) with the same molecular weight as that of the intact Tg subunit. This high molecular weight Tg was the only intralysosomal species detected by Western blot using antipig Tg antibodies. In nondenaturing conditions, lysosomal Tg (LTg) identified by radioimmunoassay was in the form of a dimer with a sedimentation coefficient lower than that of either iodinated Tg (colloid Tg) or noniodinated Tg (microsomal Tg). LTg had a lower iodine content than colloid Tg:9-12 versus 39-42 iodine atoms/molecule. Pronase hydrolysates of LTg did not contain any 3,5,3',5'-tetraiodo-L-thyronine or 3,3',5-triiodo-L-thyronine residues detectable by reverse-phase high pressure liquid chromatography; iodine present in LTg was in the form of iodotyrosines. Under reducing conditions, LTg almost completely disappeared and gave rise to various polypeptides of smaller size. These results suggest that Tg transferred to lysosomes is subjected to selective proteolytic cleavage reaction(s) that release thyroid hormone residues. This early step would lead to the formation of hormone-depleted Tg molecules that are cleaved at discrete sites, the resulting polypeptides remaining bound through disulfide bonds to yield Tg molecules with an apparently normal size and a slightly altered structure.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale Unité 197, Faculté de Médecine Alexis Carrel, Lyon, France.
authors.forename: B
authors.initials: B
authors.lastname: Rousset
authors.forename: S
authors.initials: S
authors.lastname: Selmi
authors.forename: H
authors.initials: H
authors.lastname: Bornet
authors.forename: P
authors.initials: P
authors.lastname: Bourgeat
authors.forename: R
authors.initials: R
authors.lastname: Rabilloud
authors.forename: Y
authors.initials: Y
authors.lastname: Munari-Silem
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002499:Centrifugation, Density Gradient; D002851:Chromatography, High Pressure Liquid; D008247:Lysosomes / Q000378:metabolism*; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism; D011402:Pronase; D013552:Swine; D013954:Thyroglobulin / Q000302:isolation & purification / Q000378:metabolism*; D013961:Thyroid Gland / Q000378:metabolism*; D013963:Thyroid Hormones / Q000032:analysis*
chemical_list: D000596:Amino Acids; D013963:Thyroid Hormones; D013954:Thyroglobulin; D010447:Peptide Hydrolases; D011402:Pronase