title: Analysis of the interaction between human TLR3 ectodomain and nucleic acids.
abstract: Toll-like receptor 3 (TLR3) recognizes dsRNA of viral origin and polyriboinosine-polyribocytidylic acid (poly (I:C)). TLR3 mediates the activation of IRF-3 and NF-kappaB and thereby the secretion of type I interferons and inflammatory cytokines. However, the mechanism of this activation is poorly understood. To study the molecular recognition events and biochemical interactions between TLR3 and dsRNA, human TLR3 ectodomain (ECD) fused with a signal peptide at the N-terminus and 6 x His-tag at the C-terminus (TLR3-ECD) was obtained by the baculovirus expression system. To examine the various nucleic acids binding to TLR3-ECD in vitro, a filter binding assay was carried out at pH 4.2-7.6. Interaction of TLR3-ECD with various nucleic acids (particularly dsRNA, in vitro transcripts of tRNA and HCV NS3 aptamer) required an acidic pH.
authors.affiliation: Department of Material and Biological Chemistry, Faculty of Science, Yamagata University, Yamagata 990-8560, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Fukuda
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Tsujita
authors.forename: Misako
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Seya
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakiyama
authors.forename: Fumiko
authors.initials: F
authors.lastname: Nishikawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nishikawa
authors.forename: Tsunemi
authors.initials: T
authors.lastname: Hasegawa
mesh_terms: D001665:Binding Sites; D002942:Circular Dichroism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012330:RNA, Double-Stranded / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism; D051196:Toll-Like Receptor 3 / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D012330:RNA, Double-Stranded; C495344:TLR3 protein, human; D051196:Toll-Like Receptor 3; D012343:RNA, Transfer