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Identification of iodinated proteins in cultured thyrocytes and their possible significance for thyroid hormone
P Lemansky1, G M Popp, J Tietz
1Institute of Cell Biology, Bonn, Germany.
The thyroid gland is known to generate the iodinated hormones T3 and T4 from the prohormone thyroglobulin. In this report we examined whether polypeptides other than thyroglobulin are iodinated and hormonogenic in thyrocytes and the prerequisites for their iodination. In primary cultures of porcine thyrocytes, a substantial portion of organified radioiodine was incorporated into cellular proteins other than thyroglobulin. Some of these were identified by immunoprecipitation. They included proteins of the extracellular matrix, plasma membrane proteins, and lysosomal enzymes, which follow in part a secretion and recapture pathway. All of these proteins come into contact with the iodinating system of thyrocytes located on the apical plasma membrane and possess iodination consensus sequences. Immunoprecipitation with T3- or T4-specific antibodies showed that thyroid hormones were detectable only within thyroglobulin. This was confirmed by an analysis of the iodoamino acids of thyroglobulin, cathepsin-D (representing a secretory protein), and aminopeptidase-N (a membrane-integrated protein) by two-dimensional TLC, which revealed the presence of T3 and T4 only within the polypeptide chain of thyroglobulin. These results indicate that iodoproteins other than thyroglobulin do not participate in the generation of thyroid hormones in situ.
The thyroid gland is known to generate the iodinated hormones T3 and T4 from the prohormone thyroglobulin. In this report we examined whether polypeptides other than thyroglobulin are iodinated and hormonogenic in thyrocytes and the prerequisites for their iodination. In primary cultures of porcine thyrocytes, a substantial portion of organified radioiodine was incorporated into cellular proteins other than thyroglobulin. Some of these were identified by immunoprecipitation. They included proteins of the extracellular matrix, plasma membrane proteins, and lysosomal enzymes, which follow in part a secretion and recapture pathway. All of these proteins come into contact with the iodinating system of thyrocytes located on the apical plasma membrane and possess iodination consensus sequences. Immunoprecipitation with T3- or T4-specific antibodies showed that thyroid hormones were detectable only within thyroglobulin. This was confirmed by an analysis of the iodoamino acids of thyroglobulin, cathepsin-D (representing a secretory protein), and aminopeptidase-N (a membrane-integrated protein) by two-dimensional TLC, which revealed the presence of T3 and T4 only within the polypeptide chain of thyroglobulin. These results indicate that iodoproteins other than thyroglobulin do not participate in the generation of thyroid hormones in situ.