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Evidence for a membrane carbonic anhydrase IV anchored by its C-terminal peptide in normal human pancreatic ductal cells

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Abstract

The high concentration of HCO3 − ions (150 mM) in the human pancreatic ducts raises the question of the membrane proteins responsible for their secretion in addition to the Cl−/HCO3 − exchanger. In this study, we investigated the expression of carbonic anhydrase IV (CA IV), a possible candidate. Experiments were carried out on specimens of normal human pancreas obtained from brain-dead donors (n=9) as well as on isolated human ductal cells. Two antibodies were generated: CA IV NH2 antibody directed against the NH2 terminal of human glycosyl phosphatidylinositol (GPI)-anchored CA IV and CA IV COOH antibody directed against the COOH terminal of the same protein before its association with a GPI in the rough endoplasmic reticulum. A 35-kDa CA IV was detected in the homogenates of human pancreas. Immunocytochemistry demonstrated the expression of CA IV in centroacinar cells and in intercalated, intralobular, and interlobular ductal cells. The immunoreactivity observed with the CA IV COOH antibody was mainly localized on luminal membranes of ductal cells. Treatment of purified plasma membranes with phosphatidylinositol-phospholipase C indicated that the CA IV expressed in pancreatic ducts was not GPI-anchored. Its detection in the same extracts by the CA IV COOH antibody indicated that it was anchored by a hydrophobic segment at the carboxy terminal. Taken together, these results suggest that normal human pancreatic ductal cells express a 35-kDa CA IV anchored in their luminal plasma membrane by a hydrophobic segment of the COOH terminus. In view of its localization and its mode of anchorage in luminal plasma membranes, this CA IV may participate in the maintenance of luminal pH.

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Acknowledgements

This study was financed by the “Ministère de l’Enseignement Supérieur et de la Recherche” and the French association “Vaincre la Mucoviscidose”.

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Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire et Moléculaire des Epithéliums (EA 3032), Université Paul Sabatier, 38 rue des 36 Ponts, 31400, Toulouse, France

    Marjorie Fanjul, Laetitia Alvarez, Christel Salvador & Etienne Hollande

  2. Unité INSERM U531, Biologie et Pathologie Digestive, CHU Rangueil Bât L3, 31403, Toulouse, France

    Marjorie Fanjul

  3. Cellular Therapy of Diabetes, Equipe de Recherche et d’Innovation Méthodologique, Faculté de Médecine, INSERM, 59045, Lille, France

    Valéry Gmyr, Julie Kerr-Conte & François Pattou

  4. St. George’s Hospital Medical School, Department of Child Health, University of London, Cranmer Terrace, London, SW17 ORE, UK

    Nicholas Carter

Authors
  1. Marjorie Fanjul
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  2. Laetitia Alvarez
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  3. Christel Salvador
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  4. Valéry Gmyr
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  5. Julie Kerr-Conte
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  6. François Pattou
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  7. Nicholas Carter
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  8. Etienne Hollande
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Correspondence to Etienne Hollande.

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The first two authors have contributed equally to this work

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Fanjul, M., Alvarez, L., Salvador, C. et al. Evidence for a membrane carbonic anhydrase IV anchored by its C-terminal peptide in normal human pancreatic ductal cells. Histochem Cell Biol 121, 91–99 (2004). https://doi.org/10.1007/s00418-003-0616-2

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  • DOI: https://doi.org/10.1007/s00418-003-0616-2

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