TY - JOUR
T1 - Synergistic cooperation of PDI family members in peroxiredoxin 4-driven oxidative protein folding
AU - Sato, Yoshimi
AU - Kojima, Rieko
AU - Okumura, Masaki
AU - Hagiwara, Masatoshi
AU - Masui, Shoji
AU - Maegawa, Ken Ichi
AU - Saiki, Masatoshi
AU - Horibe, Tomohisa
AU - Suzuki, Mamoru
AU - Inaba, Kenji
N1 - Funding Information:
We thank Akiko Sato for technical support. We also thank Roberto Sitia and Masakazu Kikuchi for kindly providing the expression vectors for mPrx4 and several PDIs, respectively. We are also grateful to Junichi Fujii for providing antibodies to mPrx4. This work was supported by Next Generation World-leading Researchers from MEXT to K. Inaba and by the MEXT program ‘‘X-ray Free Electron Laser Priority Strategy Program’’ and Grant-in-Aid for basic science from MEXT to M. Suzuki. The authors declare no conflicts of interest related to this work. This work was also supported by Grant-in-Aid for JSPS Fellows to M. Okumura, M. Hagiwara, and S. Masui.
Funding Information:
Crystallographic analysis. Diffraction data were collected at the Osaka University beamline BL44XU at SPring-8 equipped with an MX225-HE detector (Rayonix), whose acquisition was financially supported by the Academia Sinica and National Synchrotron Radiation Research Center (Taiwan) and at the beamline BL5A at Photon Factory equipped with an ADSC Quantum 315r detector. Data were integrated using HKL2000 software43. The crystallographic parameters are summarized in Table 1. Phase determination was made by molecular replacement using the deposited structures of human Prx4 (PDB code 2PN8) for mPrx4 C54A and P5 a0 (PDB code 3VWW) for the P5 a0–Prx4 C-terminal complex as search models with the program Phaser44. The initial structural models were refined by several cycles of manual rebuilding and refinement with Phenix.refine.
PY - 2013
Y1 - 2013
N2 - The mammalian endoplasmic reticulum (ER) harbors disulfide bond-generating enzymes, including Ero1α and peroxiredoxin 4 (Prx4), and nearly 20 members of the protein disulfide isomerase family (PDIs), which together constitute a suitable environment for oxidative protein folding. Here, we clarified the Prx4 preferential recognition of two PDI family proteins, P5 and ERp46, and the mode of interaction between Prx4 and P5 thioredoxin domain. Detailed analyses of oxidative folding catalyzed by the reconstituted Prx4-PDIs pathways demonstrated that, while P5 and ERp46 are dedicated to rapid, but promiscuous, disulfide introduction, PDI is an efficient proofreader of non-native disulfides. Remarkably, the Prx4-dependent formation of native disulfide bonds was accelerated when PDI was combined with ERp46 or P5, suggesting that PDIs work synergistically to increase the rate and fidelity of oxidative protein folding. Thus, the mammalian ER seems to contain highly systematized oxidative networks for the efficient production of large quantities of secretory proteins.
AB - The mammalian endoplasmic reticulum (ER) harbors disulfide bond-generating enzymes, including Ero1α and peroxiredoxin 4 (Prx4), and nearly 20 members of the protein disulfide isomerase family (PDIs), which together constitute a suitable environment for oxidative protein folding. Here, we clarified the Prx4 preferential recognition of two PDI family proteins, P5 and ERp46, and the mode of interaction between Prx4 and P5 thioredoxin domain. Detailed analyses of oxidative folding catalyzed by the reconstituted Prx4-PDIs pathways demonstrated that, while P5 and ERp46 are dedicated to rapid, but promiscuous, disulfide introduction, PDI is an efficient proofreader of non-native disulfides. Remarkably, the Prx4-dependent formation of native disulfide bonds was accelerated when PDI was combined with ERp46 or P5, suggesting that PDIs work synergistically to increase the rate and fidelity of oxidative protein folding. Thus, the mammalian ER seems to contain highly systematized oxidative networks for the efficient production of large quantities of secretory proteins.
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U2 - 10.1038/srep02456
DO - 10.1038/srep02456
M3 - Article
C2 - 23949117
AN - SCOPUS:84882771697
SN - 2045-2322
VL - 3
JO - Scientific Reports
JF - Scientific Reports
M1 - 2456
ER -