Structural and Dynamic Characterization of Mutated Keap1 for Varied Affinity toward Nrf2: A Molecular Dynamics Simulation Study.
Keap1 is an adaptor protein that regulates Nrf2 in response to oxidative stress. Under basal conditions, Nrf2 is negatively regulated through ubiquitination by Keap1. However, upon exposure to oxidative stress, the ubiquitination of Nrf2 is inhibited, resulting in an increased steady-state level of Nrf2 in the nucleus and increased transcription of cytoprotective genes. A gene variant G364C and somatic mutation G430C on Keap1 have recently been reported to substantially impair the Keap1-Nrf2 interaction and to be associated with lung cancer. By contrast, alanine scanning experiments have shown that the mutations S363A, S508A, S555A, and S602A do not affect the ability of Keap1 to bind to Nrf2, regardless of the fact that G364 and G430 are not in contact with Nrf2 whereas the four serine residues are involved in the accommodation of Nrf2 with their hydroxy groups. In this study, molecular dynamics simulations were performed to investigate the structural and dynamic variances among wild-type (WT) Keap1 and the six mutants in unbound form. Principal component analysis of the collected MD trajectories was performed to provide dynamic diversity. Our dynamic and structural observations suggest that the G364C and G430C mutants possess a mobile D385 that moves toward R380, an anchor residue to accommodate an acidic residue in Nrf2, thereby hampering the Keap1-Nrf2 recognition of an electrostatic nature. By contrast, none of the four serine-to-alanine mutants alters the H-bond network formed by the serine backbone to its partner; accordingly, these mutants are almost as intact as the WT structurally and dynamically.
Rehabilitation Division, Zuoying Armed Forces General Hospital , Kaohsiung, Taiwan.
I-Chung
IC
Cheng
Department of Life Sciences, National University of Kaohsiung , Kaohsiung, Taiwan.
Ya-Jyun
YJ
Chen
Department of Life Sciences, National University of Kaohsiung , Kaohsiung, Taiwan.
Chia-Wei
CW
Ku
Department of Life Sciences, National University of Kaohsiung , Kaohsiung, Taiwan.
Yu-Wen
YW
Huang
Department of Life Sciences, National University of Kaohsiung , Kaohsiung, Taiwan.
Chia-Ning
CN
Yang
D000595:Amino Acid Sequence; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000072019:Kelch-Like ECH-Associated Protein 1; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009154:Mutation; D051267:NF-E2-Related Factor 2 / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D055672:Static Electricity
D047908:Intracellular Signaling Peptides and Proteins; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human