Iodination of mature cathepsin D in thyrocytes as an indicator for its transport to the cell surface.
Thyrocytes are known for their ability to iodinate thyroglobulin from which the thyroid hormones are generated. In the intact thyroid gland the iodination process is almost exclusively executed at the apical plasma membrane of thyroid epithelial cells. Here, we show that freshly isolated thyrocytes iodinated polypeptides other than thyroglobulin and that one of the major iodinated polypeptides was the mature form of the lysosomal protease cathepsin D (CD). The detection of mature CD as an iodinated polypeptide suggested that a fraction of the lysosomally maturated enzyme was delivered to the apical plasma membrane where it became available for iodination. After labeling of thyrocytes with [35S]methionine/cysteine overnight part of the mature CD was released into the culture medium. This was abolished by inhibiting maturation of CD with NH4Cl, indicating that mature CD appeared in the medium after its proteolytic maturation in an acidic compartment. Besides CD other soluble lysosomal polypeptides like the beta-N-acetylhexosaminidase and the sphingolipid-activating protein D (Sap D) were iodinated and partially secreted as mature polypeptides. In contrast, the membrane-associated lysosomal ceramidase was iodinated and partially secreted as immature single-chain enzyme and not as fully maturated two-chain enzyme. These data indicate that a portion of mature CD and other soluble lysosomal enzymes is delivered from lysosomes to the cell surface whereas some membrane-associated enzymes from the terminal lysosomal compartment are efficiently excluded from this process.
Lemansky
P
P
Institut für Zellbiologie und Bonner Forum Biomedizin, Bonn, Germany.
Brix
K
K
Herzog
V
V
D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D002402:Cathepsin D / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D055572:Ceramidases; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007457:Iodine Radioisotopes; D015739:Nocodazole / Q000494:pharmacology; D010455:Peptides / Q000378:metabolism; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D013552:Swine; D013961:Thyroid Gland / Q000201:enzymology*; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D006023:Glycoproteins; D007457:Iodine Radioisotopes; C483345:PSAP protein, human; D010455:Peptides; D049231:Saposins; D049128:Sphingolipid Activator Proteins; D001619:beta-N-Acetylhexosaminidases; D002402:Cathepsin D; D000581:Amidohydrolases; D055572:Ceramidases; D015739:Nocodazole