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A D Dunn

Publications and source records attributed to A D Dunn.

33 records · Page 2Linked to original sources
PubMed1983

The role of iodination in the formation of hormone-rich peptides from thyroglobulin.

Reduced thyroglobulins from several animal species contain hormone-rich iodopeptides of 20,000-26,000 and 15,000-18,000 daltons. The present study has investigated the role of iodination in their production. Experimental approaches have included: iodination in vitro of thyroglobulin from rabbit thyroid slices incubated with [3H]leucine and subsequent analysis of 3H distribution by gel electrophoresis; iodination in vitro of low iodine thyroglobulin from a human goiter, followed by isolation of the major iodopeptides and quantitation of their peptide content; and injection of iodine-deficient rats with Na125I and assessment of the distribution of isotope among the iodopeptides at successive time intervals. From these experiments the following general pattern has emerged: 1) at low levels of iodine (less than 5 atoms/molecule of thyroglobulin) in vitro or at short time intervals after iodine administration in vivo (less than 4 h), the principal iodinated component of reduced thyroglobulin is a approximately 230,000-dalton peptide; 2) with moderate increases in iodine (5-40 atoms/molecule) or longer time intervals after administration (greater than or equal to 4 h) there is less of the 230,000-dalton iodopeptide, and an iodothyronine-rich approximately 20,000- to 26,000-dalton iodopeptide appears; 3) at higher levels of iodine (greater than 40 atoms/molecule) the amount of approximately 230,000-dalton iodopeptide decreases further, the amount of 20,000- to 26,000-dalton iodopeptides may decrease, and an iodothyronine-rich 15,000- to 18,000-dalton peptide appears. Progressive iodination gives the same changes in distribution of peptide material among these iodopeptides as it does in iodine distribution, and the changes seen with iodination in vitro are similar to those occurring over time in vivo. We conclude that during the process of iodination discrete peptide bonds of thyroglobulin are cleaved to produce the hormone-rich iodopeptides. This is probably a normal part of thyroglobulin maturation in vivo and may be a necessary event preceding hormone formation.

Animals↗
PubMed1982

Thyroglobulin degradation by thyroidal proteases: action of purified cathepsin D.

Cathepsin D has been purified from rabbit thyroids, and its action on thyroglobulin has been examined. The enzyme was obtained in an electrophoretically homogenous form by gel filtration, followed by ion exchange chromatography and affinity chromatography with immobilized pepstatin. In some preparations, the enzyme occurred in a high molecular weight form. The ability of cathepsin D to hydrolyze [125I]thyroglobulin to fragments with a molecular weight of less than 100K was determined by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. This activity showed a pH optimum of 3.5, was greater with reduced thyroglobulin as substrate than with the native protein, and was unaffected by potassium iodide (1-10 mM). Purified cathepsin D rapidly hydrolyzed thyroglobulin to a number of peptide intermediates. Those in the 20-45K molecular weight range had an iodothyronine content equal to or less than that of intact thyroglobulin, but the smallest peptides (apparent molecular weight, less than 2K) were iodothyronine enriched. No evidence was obtained for the release of free hormone by cathepsin D under the experimental conditions used. We conclude that cathepsin D plays a role in the initial breakdown of thyroglobulin in the thyroid and may have some selectivity for the iodothyronine portion of the molecule. The rapid hydrolysis of thyroglobulin that occurs in vivo, however, probably requires the concerted action of cathepsin D with other lysosomal endopeptidases and exopeptidases.

Animals↗
PubMed1982

Thyroglobulin degradation by thyroidal proteases: action of thiol endopeptidases in vitro.

We have obtained evidence of thiol endopeptidases in the thyroid which are active in thyroglobulin degradation in vitro. Four pepstatin-insensitive endopeptidase fractions were distinguished in extracts of rabbit thyroids by gel filtration on Bio-Gel A-0.5m. An enzyme from one fraction was obtained in highly purified form and was found to be identical to cathepsin B described in other tissues. Endopeptidases in the three remaining fractions were designated as cathepsins 180K, 110K, and 45K, respectively, on the basis of their estimated molecular size. These were partially purified by either organomercurial affinity chromatography or DEAE-cellulose chromatography. They are identified as thiol endopeptidases on the basis of their sensitivity to inhibition by both leupeptin and the thiol-blocking agent iodoacetic acid and by their activation with the reducing agent glutathione. Each is distinguished from cathepsin B on the basis of molecular size and limited ability to hydrolyze benzoylarginine-2-naphthylamide. The action of the thiol endopeptidases on [125I]thyroglobulin was analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate or in sodium dodecyl sulfate and urea. In each instance, the initial peptide fragments were approximately 40-45K and 30K, with iodothyronine contents similar to or less than that of intact thyroglobulin. Later products of digestion than that of intact thyroglobulin. Later products of digestion included first, 20K peptides, which showed a low iodothyronine content, and finally, peptides of approximately 10K, which showed a 1.5-fold enrichment of T4 and T3 over that of intact thyroglobulin. Each of the thiol endopeptidases had a synergistic effect when incubated with cathepsin D and [125I]thyroglobulin. Among the products of such incubations were small iodopeptides, which were iodothyronine-enriched, and free T4, itself. The results show that thiol endopeptidases are present in the thyroid gland and are collectively as important as cathepsin D in the hydrolysis of thyroglobulin in vitro. The action of these enzymes must be considered along with that of cathepsin D in understanding thyroglobulin hydrolysis in vivo.

Animals↗
PubMed1976

On the identity of the thyrotropic cell in the red-spotted newt.

Although the pars distalis of the red-spotted newt has previously undergone extensive cytological examination, the identity of its thyrotropic cells has remained uncertain. From the present ultrastructural study, cells of type 3 (Masur, 1969) containing granules 150-180 nm in diameter are concluded to be the thyrotropes. Such cells were found to be present in the regions of the pars distalis where thyroidectomy cells arise after ablation of the thyroid gland. Cells of type 3 are probably identical with a population of cells containing granules which stain with silver methenamine indicating the presence of a glycoprotein such as thyroid stimulating hormone (TSH). Thyroidectomy cells containing a few residual granules 150-180 nm in diameter were occasionally found in partes distales from newts killed 3 or 7 days after ablation of the thyroid gland, and were abundant in pituitaries 21 days after thyroidectomy. Only cells of type 3 responded (by vacuolation of granules) when animals were immersed in water containing 10 mug/1 of thyroxine. No cells of the pars distalis showed cytological change after administration of synthetic thyrotropic releasing hormone (TRH) giving additional support to the view that this hormone has no stimulatory role in amphibians.

Animals↗