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PMID: 21402695 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Pivotal role of the C2 domain of the Smurf1 ubiquitin ligase in substrate selection.

The Journal of biological chemistry ·Vol. 286 ·No. 19 ·2011-05-13 ·Pages 16861-70

Lu K, Li P, Zhang M, Xing G, Li X, Zhou W, Bartlam M, Zhang L, Rao Z, He F

Abstract

The C2-WW-HECT-type ubiquitin ligases Smurf1 and Smurf2 play a critical role in embryogenesis and adult bone homeostasis via regulation of bone morphogenetic protein, Wnt, and RhoA signaling pathways. The intramolecular interaction between C2 and HECT domains autoinhibits the ligase activity of Smurf2. However, the role of the Smurf1 C2 domain remains elusive. Here, we show that the C2-HECT autoinhibition mechanism is not observed in Smurf1, and instead its C2 domain functions in substrate selection. The Smurf1 C2 domain exerts a key role in localization to the plasma membrane and endows Smurf1 with differential activity toward RhoA versus Smad5 and Runx2. Crystal structure analysis reveals that the Smurf1 C2 domain possesses a typical anti-parallel β-sandwich fold. Examination of the sulfate-binding site analysis reveals two key lysine residues, Lys-28 and Lys-85, within the C2 domain that are important for Smurf1 localization at the plasma membrane, regulation on cell migration, and robust ligase activity toward RhoA, which further supports a Ca(2+)-independent localization mechanism for Smurf1. These findings demonstrate a previously unidentified role of the Smurf1 C2 domain in substrate selection and cellular localization.

MeSH Terms
Amino Acid Sequence Animals Binding Sites Calcium/chemistry Cell Line, Tumor Cell Membrane/metabolism Crystallography, X-Ray/methods Gene Expression Regulation Humans Mice Microscopy, Fluorescence/methods Molecular Sequence Data Protein Structure, Secondary Protein Structure, Tertiary Sequence Homology, Amino Acid Substrate Specificity Ubiquitin-Protein Ligases/chemistry
Chemicals
SMURF1 protein, human Ubiquitin-Protein Ligases Calcium
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Lu Kefeng
State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 100850, China.
Li Ping
Zhang Minghua
Xing Guichun
Li Xin
Zhou Weihong
Bartlam Mark
Zhang Lingqiang
Rao Zihe
He Fuchu
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
1083-351X
DOI
10.1074/jbc.M110.211979
Published
2011-05-13
Epub
2011-00-14
Pages
16861-70
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC3089529
Subset
IM
Databases
PDB
3PYC
External Links
PubMed source DOI: 10.1074/jbc.M110.211979 PMC Full Text View journal details
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