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PMID: 20172859 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Calcium activates Nedd4 E3 ubiquitin ligases by releasing the C2 domain-mediated auto-inhibition.

The Journal of biological chemistry ·Vol. 285 ·No. 16 ·2010-04-16 ·Pages 12279-88

Wang J, Peng Q, Lin Q, Childress C, Carey D, Yang W

Abstract

Nedd4 E3 ligases are members of the HECT E3 ubiquitin ligase family and regulate ubiquitination-mediated protein degradation. In this report, we demonstrate that calcium releases the C2 domain-mediated auto-inhibition in both Nedd4-1 and Nedd4-2. Calcium disrupts binding of the C2 domain to the HECT domain. Consistent with this, calcium activates the E3 ubiquitin ligase activity of Nedd4. Elevation of intracellular calcium by ionomycin treatment, or activation of acetylcholine receptor or epidermal growth factor receptor by carbachol or epidermal growth factor stimulation induced activation of endogenous Nedd4 in vivo evaluated by assays of either Nedd4 E3 ligase activity or ubiquitination of Nedd4 substrate ENaC-beta. The activation effect of calcium on Nedd4 E3 ligase activity was dramatically enhanced by a membrane-rich fraction, suggesting that calcium-mediated membrane translocation through the C2 domain might be an activation mechanism of Nedd4 in vivo. Our studies have revealed an activation mechanism of Nedd4 E3 ubiquitin ligases and established a connection of intracellular calcium signaling to regulation of protein ubiquitination.

MeSH Terms
Animals Calcium/metabolism Calcium Signaling Cell Line Endosomal Sorting Complexes Required for Transport/antagonists & inhibitors,chemistry,genetics,metabolism Enzyme Activation Epithelial Sodium Channels/metabolism Humans In Vitro Techniques Models, Biological Mutagenesis Nedd4 Ubiquitin Protein Ligases Protein Structure, Tertiary Rats Recombinant Fusion Proteins/chemistry,genetics,metabolism Sequence Deletion Substrate Specificity Transfection Ubiquitin-Protein Ligases/antagonists & inhibitors,chemistry,genetics,metabolism Ubiquitination
Chemicals
Endosomal Sorting Complexes Required for Transport Epithelial Sodium Channels Recombinant Fusion Proteins SCNN1B protein, human NEDD4L protein, rat Nedd4 Ubiquitin Protein Ligases Nedd4 protein, human Nedd4 protein, rat Nedd4L protein, human Ubiquitin-Protein Ligases Calcium
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wang Jian
Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822, PA, USA.
Peng Qisheng
Lin Qiong
Childress Chandra
Carey David
Yang Wannian
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M109.086405
Published
2010-04-16
Epub
2010-00-19
Pages
12279-88
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2852967
Subset
IM
External Links
PubMed source DOI: 10.1074/jbc.M109.086405 PMC Full Text View journal details
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