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

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Abstract

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.

Original languageEnglish
Article numbere45930
JournalPloS one
Volume7
Issue number9
DOIs
Publication statusPublished - 2012 Sep 25

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ASJC Scopus subject areas

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

Cite this

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, 7(9), [e45930]. https://doi.org/10.1371/journal.pone.0045930