Redox-dependent domain rearrangement of protein disulfide isomerase coupled with exposure of its substrate-binding hydrophobic surface

Olivier Serve, Yukiko Kamiya, Aya Maeno, Michiko Nakano, Chiho Murakami, Hiroaki Sasakawa, Yoshiki Yamaguchi, Takushi Harada, Eiji Kurimoto, Maho Yagi-Utsumi, Takeshi Iguchi, Kenji Inaba, Jun Kikuchi, Osamu Asami, Tsutomu Kajino, Toshihiko Oka, Masayoshi Nakasako, Koichi Kato

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48 Citations (Scopus)

Abstract

Protein disulfide isomerase (PDI) is a major protein in the endoplasmic reticulum, operating as an essential folding catalyst and molecular chaperone for disulfide-containing proteins by catalyzing the formation, rearrangement, and breakage of their disulfide bridges. This enzyme has a modular structure with four thioredoxin-like domains, a, b, b′, and a′, along with a C-terminal extension. The homologous a and a′ domains contain one cysteine pair in their active site directly involved in thiol-disulfide exchange reactions, while the b′ domain putatively provides a primary binding site for unstructured regions of the substrate polypeptides. Here, we report a redox-dependent intramolecular rearrangement of the b′ and a′ domains of PDI from Humicola insolens, a thermophilic fungus, elucidated by combined use of nuclear magnetic resonance (NMR) and small-angle X-ray scattering (SAXS) methods. Our NMR data showed that the substrates bound to a hydrophobic surface spanning these two domains, which became more exposed to the solvent upon oxidation of the active site of the a′ domain. The hydrogen-deuterium exchange and relaxation data indicated that the redox state of the a′ domain influences the dynamic properties of the b′ domain. Moreover, the SAXS profiles revealed that oxidation of the a′ active site causes segregation of the two domains. On the basis of these data, we propose a mechanistic model of PDI action; the a′ domain transfers its own disulfide bond into the unfolded protein accommodated on the hydrophobic surface of the substrate-binding region, which consequently changes into a "closed" form releasing the oxidized substrate.

Original languageEnglish
Pages (from-to)361-374
Number of pages14
JournalJournal of Molecular Biology
Volume396
Issue number2
DOIs
Publication statusPublished - 2010 Feb

Keywords

  • Domain rearrangement
  • Molecular chaperone
  • NMR
  • Protein disulfide isomerase
  • SAXS

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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    Serve, O., Kamiya, Y., Maeno, A., Nakano, M., Murakami, C., Sasakawa, H., Yamaguchi, Y., Harada, T., Kurimoto, E., Yagi-Utsumi, M., Iguchi, T., Inaba, K., Kikuchi, J., Asami, O., Kajino, T., Oka, T., Nakasako, M., & Kato, K. (2010). Redox-dependent domain rearrangement of protein disulfide isomerase coupled with exposure of its substrate-binding hydrophobic surface. Journal of Molecular Biology, 396(2), 361-374. https://doi.org/10.1016/j.jmb.2009.11.049