Expression and subcellular localization of a 35-kDa carbonic anhydrase IV in a human pancreatic ductal cell line (Capan-1)
| Autor: | Etienne Hollande, Marjorie Fanjul, Laetitia Alvarez |
|---|---|
| Přispěvatelé: | Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biologie Cellulaire et Moléculaire des Epithéliums, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Simon, Marie Francoise |
| Jazyk: | angličtina |
| Rok vydání: | 2007 |
| Předmět: |
Histology
MESH: Secretory Vesicles Glycosylphosphatidylinositols Ductal cells Immunoelectron microscopy Fluorescent Antibody Technique Golgi Apparatus Biology Endoplasmic Reticulum Cell Line symbols.namesake MESH: Golgi Apparatus Carbonic Anhydrase IV MESH: Endoplasmic Reticulum MESH: Microscopy Immunoelectron MESH: Pancreatic Ducts Humans Secretion Microscopy Immunoelectron MESH: Fluorescent Antibody Technique Secretory pathway MESH: Carbonic Anhydrase IV MESH: Humans Secretory Vesicles Endoplasmic reticulum Cell Membrane Pancreatic Ducts Golgi apparatus Subcellular localization MESH: Glycosylphosphatidylinositols Cell biology MESH: Cell Line Membrane symbols Anatomy MESH: Cell Membrane |
| Zdroj: | Journal of Histochemistry and Cytochemistry / Journal of Histochemistry & Cytochemistry Journal of Histochemistry and Cytochemistry / Journal of Histochemistry & Cytochemistry, 2007, 55 (8), pp.783-94. ⟨10.1369/jhc.6A7112.2007⟩ |
| DOI: | 10.1369/jhc.6A7112.2007⟩ |
| Popis: | The high intraluminal concentrations of HCO3−in the human pancreatic ducts have suggested the existence of a membrane protein supplying the Cl−/HCO3−exchanger. Membrane-bound carbonic anhydrase IV (CA IV) is one of the potential candidates for this protein. The difficulties in isolating human pancreatic ducts have led the authors to study the molecular mechanisms of HCO3−secretion in cancerous cell lines. In this work, we have characterized the CA IV expressed in Capan-1 cells. A 35-kDa CA IV was detected in cell homogenates and purified plasma membranes. Treatment of purified plasma membranes with phosphatidylinositol-phospholipase-C indicated that this CA IV was not anchored by a glycosylphosphatidylinositol (GPI). In contrast, its detection on purified plasma membranes by an antibody specifically directed against the carboxyl terminus of human immature GPI-anchored CA IV indicated that it was anchored by a C-terminal hydrophobic segment. Immunoelectron microscopy and double-labeling immunofluorescence revealed that this CA IV was present on apical plasma membranes, and in the rough endoplasmic reticulum, the endoplasmic reticulum-Golgi intermediate compartment, the Golgi complex, and secretory granules, suggesting its transport via the classical biosynthesis/secretory pathway. The expression in Capan-1 cells of a 35-kDa CA IV anchored in the apical plasma membrane through a hydrophobic segment, as is the case in the healthy human pancreas, should make the study of its role in pancreatic HCO3−secretion easier. (J Histochem Cytochem 55: 783–794, 2007) |
| Databáze: | OpenAIRE |
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