The C2 domain of the ubiquitin protein ligase Nedd4 mediates Ca2+-dependent plasma membrane localization

Détails

ID Serval
serval:BIB_33C99536737E
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
The C2 domain of the ubiquitin protein ligase Nedd4 mediates Ca2+-dependent plasma membrane localization
Périodique
Journal of Biological Chemistry
Auteur(s)
Plant  P. J., Yeger  H., Staub  O., Howard  P., Rotin  D.
ISSN
0021-9258 (Print)
Statut éditorial
Publié
Date de publication
12/1997
Volume
272
Numéro
51
Pages
32329-36
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Dec 19
Résumé
Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) is a ubiquitin protein ligase (E3) containing a hect domain, 3 or 4 WW domains, and a putative C2 domain. We have recently demonstrated an association between the WW domains of Nedd4 and the proline-rich PY motifs (XPPXY) of the epithelial Na+ channel, as well as with PY motifs of several other proteins. The role of the putative C2 domain of Nedd4 has not been elucidated. Here we show that Nedd4, endogenously expressed in Madin-Darby canine kidney cells, was redistributed from the cytosolic to the particulate fraction in response to ionomycin plus Ca2+ treatment. A similar treatment of polarized Madin-Darby canine kidney cells led to an apical and lateral membrane localization of Nedd4, as determined by immunostaining and confocal microscopy. The C2 domain of Nedd4, expressed as a glutathione S-transferase (GST) fusion protein, was sufficient to bind cellular membranes in a Ca2+-dependent manner. Moreover, this GST-Nedd4-C2 domain was able to mediate Ca2+-dependent interactions with phosphatidylserine, phosphatidylinositol, and phosphatidylcholine liposomes in vitro. An epitope-tagged Nedd4 lacking its C2 domain and stably expressed in Madin-Darby canine kidney cells failed to mediate the Ca2+-induced plasma membrane localization seen in wild-type (epitope-tagged) Nedd4. These results indicate that the putative C2 domain of Nedd4 acts as a bona fide C2 domain which binds phospholipids and membranes in a Ca2+-dependent fashion and is involved in localizing the protein primarily to the apical region of polarized epithelial cells in response to Ca2+.
Mots-clé
Amino Acid Sequence Animals Calcium/*metabolism Calcium-Binding Proteins/chemistry/genetics/*metabolism Cell Line Cell Membrane/metabolism Dogs Glutathione Transferase/metabolism *Ligases Molecular Sequence Data Rats Recombinant Fusion Proteins/metabolism Sequence Homology, Amino Acid *Ubiquitin-Protein Ligases
Pubmed
Web of science
Création de la notice
24/01/2008 14:03
Dernière modification de la notice
03/03/2018 15:49
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