Cell type-dependent differences in thyroid peroxidase cell surface expression.
Recently, it has been suggested that only approximately 2% of human thyroid peroxidase (hTPO(933)) reaches the surface of stably transfected (Chinese hamster ovary) cells, most being degraded intracellularly, and this might be representative of thyroid peroxidase (TPO) behavior in thyrocytes (Fayadat, L., Siffroi-Fernandez, S., Lanet, J., and Franc, J.-L. (2000) J. Biol. Chem. 275, 15948-15954). In agreement, in stably transfected Madin-Darby canine kidney clones, nonpermeabilized cells exhibit wild-type hTPO(933) immunofluorescence (apically) on <10% of that found in permeabilized cells, where an endoplasmic reticulum pattern is observed. Further, a C-terminally truncated, membrane-anchorless hTPO(848) is also retained in the endoplasmic reticulum of stably transfected Madin-Darby canine kidney cells. However, by contrast, in Chinese hamster ovary cells after transient transfection, hTPO(933) immunofluorescence is detected equally well in nonpermeabilized and permeabilized cells, indicating that a large portion of hTPO(933) is present at the cell surface; furthermore, hTPO(848) is efficiently secreted. Further, using an antiserum not cross-reacting with rat TPO, we find by immunofluorescence that in stable clones of PC Cl3 (rat) thyrocytes, considerably more ( approximately 50%) of the cells exhibit hTPO(933) at the cell surface. However, cell surface biotinylation and endoglycosidase H digestion assays appear to under-represent the extent of hTPO(933) transport, presumably because protein folding limits both Golgi carbohydrate modification and accessibility of lysines in the extracellular domain. We conclude that cell type-specific factors may facilitate stable expression of TPO at the cell surface of thyrocytes.
Division of Endocrinology and Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
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D000818:Animals; D019921:Biotinylation; D016466:CHO Cells; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology* / Q000378:metabolism; D002463:Cell Membrane Permeability; D006224:Cricetinae; D004285:Dogs; D004795:Enzyme Stability; D005455:Fluorescent Antibody Technique; D006596:Hexosaminidases / Q000378:metabolism; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009928:Organ Specificity; D021381:Protein Transport; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D013045:Species Specificity; D013961:Thyroid Gland / Q000166:cytology* / Q000201:enzymology* / Q000378:metabolism; D014162:Transfection
D008565:Membrane Proteins; D011994:Recombinant Proteins; D007453:Iodide Peroxidase; D006596:Hexosaminidases