Disruption of Heat Shock Protein 90 (Hsp90)-Protein Kinase Cδ (PKCδ) Interaction by (-)-Maackiain Suppresses Histamine H1 Receptor Gene Transcription in HeLa Cells.

J Biol Chem. 2015 Nov 6;290(45):27393-402.

 

Nariai Y1, Mizuguchi H2, Ogasawara T1, Nagai H1, Sasaki Y1, Okamoto Y1, Yoshimura Y3, Kitamura Y4, Nemoto H5, Takeda N4, Fukui H6.
  • 1From the Departments of Molecular Pharmacology.
  • 2From the Departments of Molecular Pharmacology, [email protected]
  • 3Clinical Pharmacy.
  • 4Otolaryngology.
  • 5Pharmaceutical Chemistry, and.
  • 6Molecular Studies for Incurable Diseases, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.

 

Abstract

The histamine H1 receptor (H1R) gene is an allergic disease sensitive gene, and its expression level is strongly correlated with the severity of allergic symptoms. (-)-Maackiain was identified as a Kujin-derived anti-allergic compound that suppresses the up-regulation of the H1R gene. However, the underlying mechanism of H1R gene suppression remains unknown. Here, we sought to identify a target protein of (-)-maackiain and investigate its mechanism of action. A fluorescence quenching assay and immunoblot analysis identified heat shock protein 90 (Hsp90) as a target protein of (-)-maackiain. A pull-down assay revealed that (-)-maackiain disrupted the interaction of Hsp90 with PKCδ, resulting in the suppression of phorbol 12-myristate 13-acetate (PMA)-induced up-regulation of H1R gene expression in HeLa cells. Additional Hsp90 inhibitors, including 17-(allylamino)-17-demethoxygeldanamycin, celastrol, and novobiocin also suppressed PMA-induced H1R gene up-regulation. 17-(Allylamino)-17-demethoxygeldanamycin inhibited PKCδ translocation to the Golgi and phosphorylation of Tyr(311) on PKCδ. These data suggest that (-)-maackiain is a novel Hsp90 pathway inhibitor. The underlying mechanism of the suppression of PMA-induced up-regulation of H1R gene expression by (-)-maackiain and Hsp90 inhibitors is the inhibition of PKCδ activation through the disruption of Hsp90-PKCδ interaction. Involvement of Hsp90 in H1R gene up-regulation suggests that suppression of the Hsp90 pathway could be a novel therapeutic strategy for allergic rhinitis.

KEYWORDS: (−)-maackiain; G protein-coupled receptor; PKCdelta; allergic disease-sensitive gene; allergy; gene expression; heat shock protein 90 (Hsp90); histamine; histamine H1 receptor gene; protein-protein interaction

PMID: 26391399