Journal
CHEMICAL BIOLOGY & DRUG DESIGN
Volume 78, Issue 6, Pages 1014-1021Publisher
WILEY
DOI: 10.1111/j.1747-0285.2011.01240.x
Keywords
Kelch ECH-associated protein 1; nuclear factor erythroid 2 (Nf-E2)-related factor 2; protein-protein interaction; solution binding assay; surface plasmon resonance
Funding
- National Institutes of Health [CA133791, CA125868]
Ask authors/readers for more resources
The Keap1Nrf2 interaction plays important roles in regulation of Nrf2 activity and induction of chemopreventive enzymes. To better understand the interaction and to determine the minimal Nrf2 sequence required for Keap1 binding, we synthesized a series of Nrf2 peptides containing ETGE motif and determined their binding affinities to the Kelch domain of Keap1 in solution using a surface plasmon resonance-based competition assay. The equilibrium dissociation constant for the interaction between 16mer Nrf2 peptide and Keap1 Kelch domain in solution (KDsoultion) was found to be 23.9 nm, which is 10X lower than the surface binding constant (KDsoultion) of 252 nm obtained for the direct binding of Keap1 Kelch domain to the immobilized 16mer Nrf2 peptide on a surface plasmon resonance sensor chip surface. The binding affinity of Nrf2 peptides to Keap1 Kelch domain was not lost until after deletion of eight residues from the N-terminus of the 16mer Nrf2 peptide. The 9mer Nrf2 peptide has a moderate binding affinity with a of 352 nm and the affinity was increased 15X upon removal of the positive charge at the peptide N-terminus by acetylation. These results suggest that the minimal Nrf2 peptide sequence required for Keap1 binding is the 9mer sequence of LDEETGEFL.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available