Subcellular distribution, secretion, and posttranslational modifications of clusterin in thyrocytes.
A prominent secretory glycoprotein was detected in the culture medium of porcine thyrocytes which was identified as clusterin by microsequencing. Treatment of thyrocytes with thyroid stimulating hormone revealed a tight regulation of both synthesis and secretion of clusterin, with a distinct fraction of clusterin being always associated with the cells. At least three N-bound glycans were found on each subunit of clusterin, receiving most of the incorporated [(32)P]phosphate-label. Binding of clusterin to the immobilized cation-independent mannose 6-phosphate (M6P) receptor indicated that part of the phosphate label was contained in M6P moieties. Immunolabeling of cultured thyrocytes and of thyrocytes in situ showed clusterin on the apical cell surfaces where it colocalized with gp330/megalin, which is known to serve as a binding site for clusterin. The association with the apical plasma membrane, which, in thyrocytes, carries the iodinating system, was confirmed by biosynthetic iodination, an as yet unknown posttranslational modification of clusterin. On the basolateral plasma membranes clusterin was found within distinct, bipartite patches, suggesting that it is a constituent of cell-adhesion complexes and that it participates in cell-cell and cell-matrix interactions.
Lemansky
P
P
Institut für Zellbiologie, Ulrich-Haberland-Strasse 61a, Bonn, 53121, Germany. lemansky@mailer.uni-marburg.de
Brix
K
K
Herzog
V
V
D000581:Amidohydrolases / Q000378:metabolism; D000595:Amino Acid Sequence; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D051152:Clusterin; D017077:Culture Media, Conditioned; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000378:metabolism*; D026541:Heymann Nephritis Antigenic Complex; D007455:Iodine / Q000378:metabolism; D008360:Mannosephosphates / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D018832:Molecular Chaperones; D008970:Molecular Weight; D009844:Oligosaccharides / Q000378:metabolism; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D017527:Receptor, IGF Type 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D013972:Thyrotropin / Q000494:pharmacology
D051152:Clusterin; D017077:Culture Media, Conditioned; D006023:Glycoproteins; D026541:Heymann Nephritis Antigenic Complex; D008360:Mannosephosphates; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D009844:Oligosaccharides; D017527:Receptor, IGF Type 2; D000643:Ammonium Chloride; C027693:mannose-6-phosphate; D013972:Thyrotropin; D007455:Iodine; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase