PDBsum entry 3wn7

Transcription

PDB id: 3wn7

Contents

Protein chains

286 a.a.

24 a.a.

Ligands

ACT ×2

Waters ×489

PDB id:

3wn7

Name:

Transcription

Title:

Crystal structure of keap1 in complex with the n-terminal region of the nrf2 transcription factor

Structure:

Kelch-like ech-associated protein 1. Chain: a, l. Fragment: keap1-dc, unp residues 321-609. Synonym: cytosolic inhibitor of nrf2, inrf2. Engineered: yes. Peptide from nuclear factor erythroid 2-related factor 2. Chain: b, m. Fragment: unp residues 17-51. Synonym: nf-e2-related factor 2, nfe2-related factor 2, nuclear

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: keap1, inrf2, kiaa0132. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes. Organism_taxid: 10090

Resolution:

1.57Å R-factor: 0.189 R-free: 0.223

Authors:

T.Fukutomi,K.Takagi,T.Mizushima,N.Ohuchi,M.Yamamoto

Key ref:

T.Fukutomi et al. (2014). Kinetic, thermodynamic, and structural characterizations of the association between Nrf2-DLGex degron and Keap1. Mol Cell Biol, 34, 832-846. PubMed id: 24366543 DOI: 10.1128/MCB.01191-13

Date:

05-Dec-13 Release date: 25-Dec-13

Protein chains

Q9Z2X8  (KEAP1_MOUSE) -  Kelch-like ECH-associated protein 1 from Mus musculus

Seq: 624 a.a.

Struc: 286 a.a.

Protein chains

Q60795  (NF2L2_MOUSE) -  Nuclear factor erythroid 2-related factor 2 from Mus musculus

Seq: 597 a.a.

Struc: 24 a.a.

DOI no: 10.1128/MCB.01191-13

Mol Cell Biol 34:832-846 (2014)

PubMed id: 24366543

Kinetic, thermodynamic, and structural characterizations of the association between Nrf2-DLGex degron and Keap1.

T.Fukutomi, K.Takagi, T.Mizushima, N.Ohuchi, M.Yamamoto.

ABSTRACT

Transcription factor Nrf2 (NF-E2-related factor 2) coordinately regulates cytoprotective gene expression, but under unstressed conditions, Nrf2 is degraded rapidly through Keap1 (Kelch-like ECH-associated protein 1)-mediated ubiquitination. Nrf2 harbors two Keap1-binding motifs, DLG and ETGE. Interactions between these two motifs and Keap1 constitute a key regulatory nexus for cellular Nrf2 activity through the formation of a two-site binding hinge-and-latch mechanism. In this study, we determined the minimum Keap1-binding sequence of the DLG motif, the low-affinity latch site, and defined a new DLGex motif that covers a sequence much longer than that previously defined. We have successfully clarified the crystal structure of the Keap1-DC-DLGex complex at 1.6 Å. DLGex possesses a complicated helix structure, which interprets well the human-cancer-derived loss-of-function mutations in DLGex. In thermodynamic analyses, Keap1-DLGex binding is characterized as enthalpy and entropy driven, while Keap1-ETGE binding is characterized as purely enthalpy driven. In kinetic analyses, Keap1-DLGex binding follows a fast-association and fast-dissociation model, while Keap1-ETGE binding contains a slow-reaction step that leads to a stable conformation. These results demonstrate that the mode of DLGex binding to Keap1 is distinct from that of ETGE structurally, thermodynamically, and kinetically and support our contention that the DLGex motif serves as a converter transmitting environmental stress to Nrf2 induction as the latch site.