MAL is an integral protein component of the machinery for apical transport in epithelial Madin–Darby canine kidney (MDCK) cells. To maintain its distribution, MAL cycles continuously between the plasma membrane and the Golgi complex. The clathrin-mediated route for apical internalization is known to differ from that at the basolateral surface. Herein, we report that MAL depends on the clathrin pathway for apical internalization. Apically internalized polymeric Ig receptor (pIgR), which uses clathrin for endocytosis, colocalized with internalized MAL in the same apical vesicles. Time-lapse confocal microscopic analysis revealed cotransport of pIgR and MAL in the same endocytic structures. Immunoelectron microscopic analysis evidenced colabeling of MAL with apically labeled pIgR in pits and clathrin-coated vesicles. Apical internalization of pIgR was abrogated in cells with reduced levels of MAL, whereas this did not occur either with its basolateral entry or the apical internalization of glycosylphosphatidylinositol-anchored proteins, which does not involve clathrin. Therefore, MAL is critical for efficient clathrin-mediated endocytosis at the apical surface in MDCK cells.
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13 October 2003
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October 06 2003
MAL regulates clathrin-mediated endocytosis at the apical surface of Madin–Darby canine kidney cells
Fernando Martín-Belmonte,
Fernando Martín-Belmonte
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
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José A. Martínez-Menárguez,
José A. Martínez-Menárguez
2Departamento de Biología Celular, Facultad de Medicina, Universidad de Murcia, Murcia, 30071 Spain
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Juan F. Aranda,
Juan F. Aranda
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
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José Ballesta,
José Ballesta
2Departamento de Biología Celular, Facultad de Medicina, Universidad de Murcia, Murcia, 30071 Spain
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María C. de Marco,
María C. de Marco
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
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Miguel A. Alonso
Miguel A. Alonso
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
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Fernando Martín-Belmonte
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
José A. Martínez-Menárguez
2Departamento de Biología Celular, Facultad de Medicina, Universidad de Murcia, Murcia, 30071 Spain
Juan F. Aranda
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
José Ballesta
2Departamento de Biología Celular, Facultad de Medicina, Universidad de Murcia, Murcia, 30071 Spain
María C. de Marco
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
Miguel A. Alonso
1Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain
Address correspondence to Miguel A. Alonso, Centro de Biología Molecular “Severo Ochoa”, Universidad Autónoma de Madrid and Consejo Superior de Investigaciones Científicas, Cantoblanco, Madrid, 28049 Spain. Tel.: 34-91-397-8037. Fax: 34-91-397-8087. email: [email protected]
The online version of this article contains supplemental material.
Abbreviations used in this paper: FR, folate receptor; GPI, glycosylphosphatidylinositol; NHS-SS-biotin, sulfosuccinimidyl 2-(6-(biotinamido)hexanoamido)ethyl-1-3′-dithiopropionate; pIgR, polymeric Ig receptor; sulfo-NHS-biotin, sulfo-N-hydroxyl-succinimido-biotin.
Received:
April 10 2003
Accepted:
August 14 2003
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2003
J Cell Biol (2003) 163 (1): 155–164.
Article history
Received:
April 10 2003
Accepted:
August 14 2003
Citation
Fernando Martín-Belmonte, José A. Martínez-Menárguez, Juan F. Aranda, José Ballesta, María C. de Marco, Miguel A. Alonso; MAL regulates clathrin-mediated endocytosis at the apical surface of Madin–Darby canine kidney cells . J Cell Biol 13 October 2003; 163 (1): 155–164. doi: https://doi.org/10.1083/jcb.200304053
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