title: Disruption of Heat Shock Protein 90 (Hsp90)-Protein Kinase Cδ (PKCδ) Interaction by (-)-Maackiain Suppresses Histamine H1 Receptor Gene Transcription in HeLa Cells.
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.
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Nariai
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan. Electronic address: guchi003@tokushima-u.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mizuguchi
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Takeyasu
authors.initials: T
authors.lastname: Ogasawara
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nagai
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Yohei
authors.initials: Y
authors.lastname: Sasaki
authors.affiliation: Departments of Molecular Pharmacology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Okamoto
authors.affiliation: Departments of Clinical Pharmacy, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Yoshimura
authors.affiliation: Departments of Otolaryngology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kitamura
authors.affiliation: Departments of Pharmaceutical Chemistry, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Hisao
authors.initials: H
authors.lastname: Nemoto
authors.affiliation: Departments of Otolaryngology, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Takeda
authors.affiliation: Departments of Molecular Studies for Incurable Diseases, Institute of Biomedical Sciences, Tokushima University Graduate School, Tokushima 770-8505, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Fukui
mesh_terms: D018926:Anti-Allergic Agents / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000378:metabolism; D018833:Chaperonins / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006634:Histamine H1 Antagonists / Q000494:pharmacology; D006801:Humans; D015847:Interleukin-4 / Q000235:genetics; D053978:Pentacyclic Triterpenes; D010766:Phosphorylation; D011485:Protein Binding; D051745:Protein Kinase C-delta / Q000378:metabolism*; D036343:Pterocarpans / Q000494:pharmacology*; D011794:Quercetin / Q000494:pharmacology; D011969:Receptors, Histamine H1 / Q000235:genetics*; D015398:Signal Transduction / Q000187:drug effects; D014158:Transcription, Genetic / Q000187:drug effects; D014315:Triterpenes / Q000494:pharmacology; D000094162:Phytoalexins
chemical_list: D018926:Anti-Allergic Agents; D018797:Cell Cycle Proteins; D018833:Chaperonins; D018841:HSP90 Heat-Shock Proteins; D006634:Histamine H1 Antagonists; D015847:Interleukin-4; D053978:Pentacyclic Triterpenes; D051745:Protein Kinase C-delta; D036343:Pterocarpans; D011794:Quercetin; D011969:Receptors, Histamine H1; D014315:Triterpenes; C498362:CDC37 protein, human; C508598:IL4 protein, human; C050414:celastrol; C001449:inermin; D000094162:Phytoalexins
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