title: Mitochondrial proteins NIP-SNAP-1 and -2 are a target for the immunomodulatory activity of clarithromycin, which involves NF-κB-mediated cytokine production.
abstract: Macrolide antibiotics have immunomodulatory activities, including suppression of cytokine production, cell adhesion molecule expression, and mucin production. These immunomodulatory activities improve the symptoms of respiratory diseases associated with chronic inflammation. However, the underlying molecular mechanism(s) is not well understood yet. To address this, we prepared clarithromycin (CAM)-conjugated Sepharose and examined bound cellular proteins by proteome analysis. We identified mitochondrial proteins 4-nitrophenylphosphatase domain and non-neuronal synaptosomal associated protein 25-like protein homolog (NIP-SNAP)-1 and -2 and very long-chain acyl-CoA dehydrogenase (VLCAD) as CAM-binding proteins. Production of proinflammatory cytokines (IL-8 and IL-6) induced by lipopolysaccharides (LPSs) and Pam3-CSK4in human epithelial cell lines BEAS-2B and T24 were suppressed by knockdown of NIP-SNAP-1 or -2, and partly by knockdown of VLCAD. Also, knockdown of NIP-SNAP-1 or -2 in various cell lines suppressed LPS-induced expression of IL-8 and IL-6 mRNA and NF-κB activity. Thus, CAM suppresses NF-κB-mediated proinflammatory cytokine production by interacting with mitochondrial proteins, NIP-SNAP-1 and -2.
authors.affiliation: Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Soh
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan; Department of Otorhinolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Ogasawara
authors.affiliation: Department of Otorhinolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Yamamoto
authors.affiliation: Faculty of Pharmacy, Meijo University, Naogya, Japan.
authors.forename: Chika
authors.initials: C
authors.lastname: Uemura
authors.affiliation: Faculty of Pharmacy, Meijo University, Naogya, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Takaya
authors.affiliation: Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Shiraishi
authors.affiliation: Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Toyotaka
authors.initials: T
authors.lastname: Sato
authors.affiliation: Department of Pediatrics, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Hashimoto
authors.affiliation: Department of Pediatrics, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tsutsumi
authors.affiliation: Department of Otorhinolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Takano
authors.affiliation: Department of Otorhinolaryngology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Himi
authors.affiliation: Department of Microbiology, Sapporo Medical University School of Medicine, Sapporo, Japan. Electronic address: syokota@sapmed.ac.jp.
authors.forename: Shin-Ichi
authors.initials: SI
authors.lastname: Yokota
mesh_terms: D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D017291:Clarithromycin / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides / Q000494:pharmacology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D010750:Phosphoproteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism; D051193:Toll-Like Receptors / Q000819:agonists
chemical_list: D016207:Cytokines; D007155:Immunologic Factors; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D016328:NF-kappa B; C113197:NIPSNAP1 protein, human; C113390:NIPSNAP2 protein, human; D010750:Phosphoproteins; D011506:Proteins; D012333:RNA, Messenger; C494208:SNAP25 protein, human; D050825:Synaptosomal-Associated Protein 25; D051193:Toll-Like Receptors; D044942:Acyl-CoA Dehydrogenase, Long-Chain; D017291:Clarithromycin
keywords: Clarithromycin; Innate immunity; Macrolide antibiotics; Mitochondria; NIP-SNAP; Proinflammatory cytokines