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PMID: 17785452 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Different electrostatic potentials define ETGE and DLG motifs as hinge and latch in oxidative stress response.

Molecular and cellular biology ·Vol. 27 ·No. 21 ·2007-11-00 ·Pages 7511-21

Tong KI, Padmanabhan B, Kobayashi A, Shang C, Hirotsu Y, Yokoyama S, Yamamoto M

Abstract

Nrf2 is the regulator of the oxidative/electrophilic stress response. Its turnover is maintained by Keap1-mediated proteasomal degradation via a two-site substrate recognition mechanism in which two Nrf2-Keap1 binding sites form a hinge and latch. The E3 ligase adaptor Keap1 recognizes Nrf2 through its conserved ETGE and DLG motifs. In this study, we examined how the ETGE and DLG motifs bind to Keap1 in a very similar fashion but with different binding affinities by comparing the crystal complex of a Keap1-DC domain-DLG peptide with that of a Keap1-DC domain-ETGE peptide. We found that these two motifs interact with the same basic surface of either Keap1-DC domain of the Keap1 homodimer. The DLG motif works to correctly position the lysines within the Nrf2 Neh2 domain for efficient ubiquitination. Together with the results from calorimetric and functional studies, we conclude that different electrostatic potentials primarily define the ETGE and DLG motifs as a hinge and latch that senses the oxidative/electrophilic stress.

MeSH Terms
Adaptor Proteins, Signal Transducing/chemistry,metabolism Amino Acid Motifs Amino Acid Sequence Amino Acids/metabolism Animals Calorimetry Cell Line Crystallography, X-Ray Cytoskeletal Proteins/chemistry,metabolism Humans Kelch-Like ECH-Associated Protein 1 Mice Molecular Sequence Data Mutant Proteins/metabolism Mutation/genetics NF-E2-Related Factor 2/metabolism Oxidative Stress/physiology Peptides/chemistry,metabolism Protein Binding Protein Structure, Tertiary Static Electricity Structure-Activity Relationship Thermodynamics Ubiquitin/metabolism
Chemicals
Adaptor Proteins, Signal Transducing Amino Acids Cytoskeletal Proteins Keap1 protein, mouse Kelch-Like ECH-Associated Protein 1 Mutant Proteins NF-E2-Related Factor 2 Peptides Ubiquitin
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Tong Kit I
Graduate School of Comprehensive Human Sciences, Center for TARA, JST-ERATO Environmental Response Project, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba 305-8577, Japan.
Padmanabhan Balasundaram
Kobayashi Akira
Shang Chengwei
Hirotsu Yosuke
Yokoyama Shigeyuki
Yamamoto Masayuki
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.00753-07
Published
2007-11-00
Epub
2007-00-04
Pages
7511-21
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC2169061
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.00753-07 PMC Full Text View journal details
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