@article{ad6558d676d14bc788b413ef79a505a9,
title = "A unique leucine-valine adhesive motif supports structure and function of protein disulfide isomerase P5 via dimerization",
abstract = "P5, also known as PDIA6, is a PDI family member involved in the ER quality control. Here, we revealed that P5 dimerizes via a unique adhesive motif contained in the N-terminal thioredoxin-like domain. Unlike conventional leucine zipper motifs with leucine residues every two helical turns on ∼30-residue parallel α helices, this adhesive motif includes periodic repeats of leucine/valine residues at the third or fourth position spanning five helical turns on 15-residue anti-parallel α helices. The P5 dimerization interface is further stabilized by several reciprocal salt bridges and C-capping interactions between protomers. A monomeric P5 mutant with the impaired adhesive motif showed structural instability and local unfolding, and behaved as aberrant proteins that induce the ER stress response. Disassembly of P5 to monomers compromised its ability to inactivate IRE1α via intermolecular disulfide bond reduction and its Ca2+-dependent regulation of chaperone function in vitro. Thus, the leucine-valine adhesive motif supports structure and function of P5.",
keywords = "calcium binding, dimerization motif, ER quality control, NMR, oxidative protein folding, P5, protein disulfide isomerase, SAXS",
author = "Masaki Okumura and Shingo Kanemura and Motonori Matsusaki and Misaki Kinoshita and Tomohide Saio and Dai Ito and Chihiro Hirayama and Hiroyuki Kumeta and Mai Watabe and Yuta Amagai and Lee, {Young Ho} and Shuji Akiyama and Kenji Inaba",
note = "Funding Information: Synchrotron radiation experiments were performed on SPring-8 beamline BL45XU with the approval of RIKEN (proposal nos. 2014A1345 , 2017B1176 , 2018A1311 , and 2018B1457 ). This work was partly supported by the Nanotechnology Platform Program Molecule and Material Synthesis ( JPMXP09S20MS0001 ) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. We are grateful to N. Fukamachi (Tohoku University) for experimental assistance. We are grateful to Dr. Y. Lin from KBSI for ITC analysis. We thank Dr. Kozo Tanaka (Tohoku University) for providing HeLa Kyoto cells. This research was funded by JSPS KAKENHI grant no. JP17H06521 (to S.K.), JP19K16092 (to S.K.), JP19J00893 (to M.M.), and JP20K15969 (to M.M.). We acknowledge, with thanks, funding from a Grant-in-Aid for Scientific Research on Innovative Areas from MEXT ( 19H04799 and 20H04688 to M.O.), the Takeda Science Foundation (to K.I. and M.O.), the Mochida Memorial Foundation for Medical and Pharmaceutical Research (to M.O.), the Japan Foundation of Applied Enzymology (to M.O.), the Building of Consortia for the Development of Human Resources in Science and Technology (to M.O.), a Grant-in-Aid for Scientific Research (C) to M.O. ( 19K06520 ), and Scientific Research (A) to K.I. ( 18H03978 ), the Promotion of Joint International Research (Fostering Joint International Research; B) to T.S., M.O., and S.K. ( 20345793 ), Ensemble Grants for Early Career Researchers 2020 to M.M. and M.O., National Research Foundation of Korea (NRF) grants funded by the Korean Government ( NRF-2018K1A3A1A39088040 and NRF-2019R1A2C1004954 ) to Y.-H.L., a National Research Council of Science & Technology (NST) grant funded by the Korea Government (MSIP; CAP-17-05-KIGAM) to Y.-H.L., and a Korea Basic Science Institute grant ( C070410 ) to Y.-H.L. Funding Information: Synchrotron radiation experiments were performed on SPring-8 beamline BL45XU with the approval of RIKEN (proposal nos. 2014A1345, 2017B1176, 2018A1311, and 2018B1457). This work was partly supported by the Nanotechnology Platform Program Molecule and Material Synthesis (JPMXP09S20MS0001) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. We are grateful to N. Fukamachi (Tohoku University) for experimental assistance. We are grateful to Dr. Y. Lin from KBSI for ITC analysis. We thank Dr. Kozo Tanaka (Tohoku University) for providing HeLa Kyoto cells. This research was funded by JSPS KAKENHI grant no. JP17H06521 (to S.K.), JP19K16092 (to S.K.), JP19J00893 (to M.M.), and JP20K15969 (to M.M.). We acknowledge, with thanks, funding from a Grant-in-Aid for Scientific Research on Innovative Areas from MEXT (19H04799 and 20H04688 to M.O.), the Takeda Science Foundation (to K.I. and M.O.), the Mochida Memorial Foundation for Medical and Pharmaceutical Research (to M.O.), the Japan Foundation of Applied Enzymology (to M.O.), the Building of Consortia for the Development of Human Resources in Science and Technology (to M.O.), a Grant-in-Aid for Scientific Research (C) to M.O. (19K06520), and Scientific Research (A) to K.I. (18H03978), the Promotion of Joint International Research (Fostering Joint International Research; B) to T.S. M.O. and S.K. (20345793), Ensemble Grants for Early Career Researchers 2020 to M.M. and M.O. National Research Foundation of Korea (NRF) grants funded by the Korean Government (NRF-2018K1A3A1A39088040 and NRF-2019R1A2C1004954) to Y.-H.L. a National Research Council of Science & Technology (NST) grant funded by the Korea Government (MSIP; CAP-17-05-KIGAM) to Y.-H.L. and a Korea Basic Science Institute grant (C070410) to Y.-H.L. M.O. designed and performed parts of experiments, including SAXS and oxidative protein folding experiments. S.K. performed SAXS and chaperone activity experiments. M.M. prepared purified IRE1 LD, and performed reduction assays and P5 overexpression experiments in cultured cells. M.K. D.I. and Y.-H.L. performed ITC and CD experiments. T.S. and H.K. performed NMR experiments. C.H. and M.W. prepared P5 and P5 mutant proteins. Y.A. assisted with cell culture experiments. S.A. analyzed the SAXS data. K.I. supervised the study. K.I. and M.O. wrote the manuscript. M.O. and K.I. prepared figures. All authors discussed the results and approved the manuscript. The authors declare no conflict of interest. Publisher Copyright: {\textcopyright} 2021 Elsevier Ltd",
year = "2021",
month = dec,
day = "2",
doi = "10.1016/j.str.2021.03.016",
language = "English",
volume = "29",
pages = "1357--1370.e6",
journal = "Structure",
issn = "0969-2126",
publisher = "Cell Press",
number = "12",
}