Polarized distribution and delivery of plasma membrane proteins in thyroid follicular epithelial cells.
Thyroid follicular cells coordinate several oppositely located surface enzyme activities. Recent studies have raised questions about the basic mechanisms used to achieve thyroid surface polarity. We investigated these mechanisms in primary thyroid epithelial monolayers cultured on porous filters. In the steady state, most Na+/K(+)-ATpase and aminopeptidase N were available for surface biotinylation, and these proteins exhibited physiological distributions (basolateral and apical, respectively). Glycosylphosphatidylinositol-anchored proteins were also apically distributed. By pulse-chase, newly synthesized transmembrane proteins exhibited polarized surface delivery that was oriented similarly to that observed at steady state. Little time elapsed between acquisition of Golgi-specific processing and cell surface arrival. Interestingly, when either newly synthesized or steady state-labeled thyroid peroxidase was similarly analyzed, only approximately 30% of the enzyme was ever detected at the cell surface. Of this, the majority was localized apically. The data suggest that most thyroid peroxidase remains intracellular in these monolayers, consistent with the possibility of intracellular iodination activity in addition to apical extracellular iodination. Nevertheless, in filter-polarized thyrocytes, most newly synthesized plasma membrane proteins appear to be sorted in the Golgi complex for direct delivery to apical and basolateral domains.
Division of Endocrinology, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts.
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R
Kuliawat
M P
MP
Lisanti
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P
Arvan
D000818:Animals; D018826:CD13 Antigens / Q000378:metabolism; D016764:Cell Polarity; D002478:Cells, Cultured; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D007453:Iodide Peroxidase / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013552:Swine; D013961:Thyroid Gland / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*
D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D007453:Iodide Peroxidase; D018826:CD13 Antigens; D000254:Sodium-Potassium-Exchanging ATPase