Dual targeting of Keap1 and Gsk-3 by hexaraphane in the regulation of transcription factor Nrf2

6-(Methylsulfinyl)hexyl isothiocyanate, here referred to as hexaraphane, is a bioactive compound found in wasabi, which exhibits cytoprotective, anti-inflammatory, and chemopreventive properties. The beneficial effects of hexaraphane are largely mediated through derepression of transcription factor...

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Detalles Bibliográficos
Autores: García Yagüe, Ángel Juan, Cueto Díaz, Eduardo J., Escoll, Maribel, Okunishi, Isao, Hayes, John D., Rodríguez Franco, María Isabel, Rojo Sanchís, Ana Isabel, Cuadrado Pastor, Antonio
Tipo de recurso: artículo
Fecha de publicación:2025
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/728520
Acceso en línea:https://hdl.handle.net/10486/728520
https://dx.doi.org/10.1016/j.freeradbiomed.2025.07.051
Access Level:acceso abierto
Palabra clave:Hexaraphane
Nrf2
Keap1
Gsk-3β
Molecular docking
Molecular dynamics
Medicina
Química
Descripción
Sumario:6-(Methylsulfinyl)hexyl isothiocyanate, here referred to as hexaraphane, is a bioactive compound found in wasabi, which exhibits cytoprotective, anti-inflammatory, and chemopreventive properties. The beneficial effects of hexaraphane are largely mediated through derepression of transcription factor Nrf2, which is typically repressed via proteasomal degradation mediated by its two-site interaction with the ubiquitin E3 ligase adapter Keap1. Like other isothiocyanates, hexaraphane increases Nrf2 activity by perturbing the Nrf2–Keap1 interaction and stalling Keap1-directed ubiquitination. However, we found that hexaraphane modestly increases Nrf2 levels in Keap1-deficient cells, suggesting an additional Keap1-independent mechanism. Unlike other electrophilic Nrf2 activators, hexaraphane did not significantly impact Erk, p38Mapk, Jnk, or Pten/Pi3k/Akt signaling. Instead, our data reveal that hexaraphane inhibits Gsk-3β, a kinase that targets Nrf2 for proteasomal degradation by phosphorylating a DSGIS degron in the transcription factor through which it interacts with the β-TrCP E3 ligase adaptor. Hexaraphane reduced Nrf2 ubiquitination and its binding to Gsk-3β, mimicking the effects of the Gsk-3β inhibitor SB216763. In vitro kinase assays confirmed that hexaraphane suppresses Nrf2 phosphorylation. Furthermore, simulations of molecular docking and dynamics predict a specific interaction between hexaraphane and the catalytic groove of Gsk-3β. These findings identify hexaraphane as a previously unrecognized dual-acting Nrf2 activator that stabilizes Nrf2 both by disturbing Keap1 binding and by inhibiting Gsk-3β that forms the DSGIpS338 phosphodegron