NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47

Maho Yagi-Utsumi, Sumi Yoshikawa, Yoshiki Yamaguchi, Yohei Nishi, Eiji Kurimoto, Yoshihito Ishida, Takayuki Homma, Jun Hoseki, Yoshimi Nishikawa, Takaki Koide, Kazuhiro Nagata, Koichi Kato

    研究成果: Article

    10 引用 (Scopus)

    抄録

    Heat shock protein 47 (Hsp47) acts as a client-specific chaperone for collagen and plays a vital role in collagen maturation and the consequent embryonic development. In addition, this protein can be a potential target for the treatment of fibrosis. Despite its physiological and pathological importance, little is currently known about the collagen-binding mode of Hsp47 from a structural aspect. Here, we describe an NMR study that was conducted to identify the collagen-binding site of Hsp47. We used chicken Hsp47, which has higher solubility than its human counterpart, and applied a selective 15N-labeling method targeting its tryptophan and histidine residues. Spectral assignments were made based on site-directed mutagenesis of the individual residues. By inspecting the spectral changes that were observed upon interaction with a trimeric collagen peptide and the mutational data, we successfully mapped the collagen-binding site in the B/C β-barrel domain and a nearby loop in a 3D-homology model based upon a serpin fold. This conclusion was confirmed by mutational analysis. Our findings provide a molecular basis for the design of compounds that target the interaction between Hsp47 and procollagen as therapeutics for fibrotic diseases.

    元の言語English
    記事番号e45930
    ジャーナルPLoS One
    7
    発行部数9
    DOI
    出版物ステータスPublished - 2012 9 25

    Fingerprint

    HSP47 Heat-Shock Proteins
    heat shock proteins
    collagen
    binding sites
    Collagen
    Binding Sites
    Nuclear magnetic resonance
    Serpins
    Procollagen
    Mutagenesis
    site-directed mutagenesis
    Site-Directed Mutagenesis
    fibrosis
    histidine
    Histidine
    Tryptophan
    tryptophan
    Solubility
    Labeling
    Embryonic Development

    ASJC Scopus subject areas

    • Agricultural and Biological Sciences(all)
    • Biochemistry, Genetics and Molecular Biology(all)
    • Medicine(all)

    これを引用

    Yagi-Utsumi, M., Yoshikawa, S., Yamaguchi, Y., Nishi, Y., Kurimoto, E., Ishida, Y., ... Kato, K. (2012). NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47. PLoS One, 7(9), [e45930]. https://doi.org/10.1371/journal.pone.0045930

    NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47. / Yagi-Utsumi, Maho; Yoshikawa, Sumi; Yamaguchi, Yoshiki; Nishi, Yohei; Kurimoto, Eiji; Ishida, Yoshihito; Homma, Takayuki; Hoseki, Jun; Nishikawa, Yoshimi; Koide, Takaki; Nagata, Kazuhiro; Kato, Koichi.

    :: PLoS One, 巻 7, 番号 9, e45930, 25.09.2012.

    研究成果: Article

    Yagi-Utsumi, M, Yoshikawa, S, Yamaguchi, Y, Nishi, Y, Kurimoto, E, Ishida, Y, Homma, T, Hoseki, J, Nishikawa, Y, Koide, T, Nagata, K & Kato, K 2012, 'NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47', PLoS One, 巻. 7, 番号 9, e45930. https://doi.org/10.1371/journal.pone.0045930
    Yagi-Utsumi M, Yoshikawa S, Yamaguchi Y, Nishi Y, Kurimoto E, Ishida Y その他. NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47. PLoS One. 2012 9 25;7(9). e45930. https://doi.org/10.1371/journal.pone.0045930
    Yagi-Utsumi, Maho ; Yoshikawa, Sumi ; Yamaguchi, Yoshiki ; Nishi, Yohei ; Kurimoto, Eiji ; Ishida, Yoshihito ; Homma, Takayuki ; Hoseki, Jun ; Nishikawa, Yoshimi ; Koide, Takaki ; Nagata, Kazuhiro ; Kato, Koichi. / NMR and Mutational Identification of the Collagen-Binding Site of the Chaperone Hsp47. :: PLoS One. 2012 ; 巻 7, 番号 9.
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