TY - JOUR
T1 - Radically different thioredoxin domain arrangement of ERp46, an efficient disulfide bond introducer of the mammalian PDI family
AU - Kojima, Rieko
AU - Okumura, Masaki
AU - Masui, Shoji
AU - Kanemura, Shingo
AU - Inoue, Michio
AU - Saiki, Masatoshi
AU - Yamaguchi, Hiroshi
AU - Hikima, Takaaki
AU - Suzuki, Mamoru
AU - Akiyama, Shuji
AU - Inaba, Kenji
N1 - Funding Information:
This work was supported by a grant for Next Generation World-leading Researchers from MEXT to K.I., the MEXT program “X-ray Free Electron Laser Priority Strategy Program” to M.S., MEXT to S.A., and a Grant-in-Aid for JSPS Fellows to M.O. and S.M. The synchrotron radiation experiments were performed at BL45XU in SPring-8 with approval of RIKEN (proposal no. 2013A1113).
Funding Information:
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). For structural determination of ERp46 Trx1 and Trx2, data were integrated with IMOSFLM ( Leslie, 2006 ) and scaled with SCALA from the CCP4 interface ( CCP4, 1994 ). Phase determination was made by molecular replacement using the deposited structure of ERp46 Trx3 (PDB code 3UVT ) as a search model with the program Phaser ( McCoy et al., 2007 ). The structures were manually rebuilt with Coot ( Emsley and Cowtan, 2004 ) and refined with Refmac ( Murshudov et al., 1997 ) and phenix.refine. Data collection and refinement statistics are given in Table 1 . For structural determination of the ERp46 Trx2-Prx4 C-terminal peptide complex, data were integrated using the HKL2000 software ( Otwinowski and Minor, 1997 ). Phase determination was made by molecular replacement using Phaser, with ERp46 Trx2 (PDB code 3WGE) as a search model. The initial structural models were refined by several cycles of manual rebuilding and refinement with phenix.refine. Figures depicting the structure were prepared with the program PyMOL (Delano Scientific).
PY - 2014/3/4
Y1 - 2014/3/4
N2 - The mammalian endoplasmic reticulum (ER) contains a diverse oxidative protein folding network in which ERp46, a member of the protein disulfide isomerase (PDI) family, serves as an efficient disulfide bond introducer together with Peroxiredoxin-4 (Prx4). We revealed a radically different molecular architecture of ERp46, in which the N-terminal two thioredoxin (Trx) domains with positively charged patches near their peptide-binding site and the C-terminal Trx are linked by unusually long loops and arranged extendedly, forming an opened V-shape. Whereas PDI catalyzes native disulfide bond formation by the cooperative action of two mutually facing redox-active sites on folding intermediates bound to the central cleft, ERp46 Trx domains are separated, act independently, and engage in rapid but promiscuous disulfide bond formation during early oxidative protein folding. Thus, multiple PDI family members likely contribute to different stages of oxidative folding and work cooperatively to ensure the efficient production of multi-disulfide proteins in the ER.
AB - The mammalian endoplasmic reticulum (ER) contains a diverse oxidative protein folding network in which ERp46, a member of the protein disulfide isomerase (PDI) family, serves as an efficient disulfide bond introducer together with Peroxiredoxin-4 (Prx4). We revealed a radically different molecular architecture of ERp46, in which the N-terminal two thioredoxin (Trx) domains with positively charged patches near their peptide-binding site and the C-terminal Trx are linked by unusually long loops and arranged extendedly, forming an opened V-shape. Whereas PDI catalyzes native disulfide bond formation by the cooperative action of two mutually facing redox-active sites on folding intermediates bound to the central cleft, ERp46 Trx domains are separated, act independently, and engage in rapid but promiscuous disulfide bond formation during early oxidative protein folding. Thus, multiple PDI family members likely contribute to different stages of oxidative folding and work cooperatively to ensure the efficient production of multi-disulfide proteins in the ER.
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U2 - 10.1016/j.str.2013.12.013
DO - 10.1016/j.str.2013.12.013
M3 - Article
C2 - 24462249
AN - SCOPUS:84896717675
VL - 22
SP - 431
EP - 443
JO - Structure with Folding & design
JF - Structure with Folding & design
SN - 0969-2126
IS - 3
ER -