Studies of the congenitally goitrous sheep. Iodoproteins of the goitre

Abstract

1. Congenitally goitrous thyroid tissue was obtained from South Australian Merino sheep. Ultrastructural studies of the secretory cells in this tissue showed active cells of normal appearance, containing apical protein droplets. 2. ¹²⁵I-labelling in vivo of goitre tissue was used to investigate the iodoproteins, in which the major proportion of ¹²⁵I appeared in the cell protein fraction soluble in 0.9% sodium chloride (average 62% in goitres from untreated sheep). 3. Ammonium sulphate fractionation showed two clear peaks of iodoprotein precipitation, one at 35–40% saturation and the other at 50–55% saturation. Both iodoprotein fractions contained iodotyrosines and iodothyronines, which were identified chromatographically after enzymic hydrolysis of the protein. 4. Polyacrylamide-gel electrophoresis at pH9.4, at either 7.5 or 5.0% acrylamide concentration, was used to characterize the iodoproteins. Two major fractions were observed, the fastest-migrating fraction coincident with serum albumin, and a slower-migrating, less-well-defined zone. This fraction migrated in 7.5% acrylamide gel, which excluded normal thyroglobulin. 5. Density-gradient (10–40% sucrose) centrifugation was used to determine the approximate sedimentation coefficients of the iodoproteins, which showed major components at s_20,w 8–9S and s_20,w125I-labelled proteins from goitre extracts. 7. Ouchterlony-type double diffusion in agar plates demonstrated immunoprecipitation lines between rabbit anti-(sheep thyroglobulin) and both the concentrated goitre extract and its Sephadex G-200-excluded fraction, which were confluent with that obtained on reaction with purified normal thyroglobulin. 8. It was concluded that both major iodoprotein fractions were capable of supplying thyroid hormones to the animal, and that the fraction of s_20,w125I-labelled thyroglobulin was not detected in goitre tissue from untreated or thyroxine-treated animals, it was possible that the genetic defect causing goitre resulted in an abnormal thyroglobulin, incapable of being iodinated but immunologically reactive.