title: Cooperative Protein Folding by Two Protein Thiol Disulfide Oxidoreductases and 1 in Soybean.
abstract: Most proteins produced in the endoplasmic reticulum (ER) of eukaryotic cells fold via disulfide formation (oxidative folding). Oxidative folding is catalyzed by protein disulfide isomerase (PDI) and PDI-related ER protein thiol disulfide oxidoreductases (ER oxidoreductases). In yeast and mammals, ER oxidoreductin-1s (Ero1s) supply oxidizing equivalent to the active centers of PDI. In this study, we expressed recombinant soybean Ero1 (GmERO1a) and found that GmERO1a oxidized multiple soybean ER oxidoreductases, in contrast to mammalian Ero1s having a high specificity for PDI. One of these ER oxidoreductases, GmPDIM, associated in vivo and in vitro with GmPDIL-2, was unable to be oxidized by GmERO1a. We therefore pursued the possible cooperative oxidative folding by GmPDIM, GmERO1a, and GmPDIL-2 in vitro and found that GmPDIL-2 synergistically accelerated oxidative refolding. In this process, GmERO1a preferentially oxidized the active center in the A': domain among the A: , A': , and B: domains of GmPDIM. A disulfide bond introduced into the active center of the A': domain of GmPDIM was shown to be transferred to the active center of the A: domain of GmPDIM and the A: domain of GmPDIM directly oxidized the active centers of both the A: or A': domain of GmPDIL-2. Therefore, we propose that the relay of an oxidizing equivalent from one ER oxidoreductase to another may play an essential role in cooperative oxidative folding by multiple ER oxidoreductases in plants.
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Motonori
authors.identifier: http://orcid.org/0000-0002-7296-033X
authors.initials: M
authors.lastname: Matsusaki
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Okuda
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Masuda
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Koishihara
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Ryuta
authors.initials: R
authors.lastname: Mita
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Kensuke
authors.initials: K
authors.lastname: Iwasaki
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Hara
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Yurika
authors.initials: Y
authors.lastname: Naruo
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Akiho
authors.initials: A
authors.lastname: Hirose
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Tsuchi
authors.affiliation: Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan urade@kais.kyoto-u.ac.jp.
authors.forename: Reiko
authors.identifier: http://orcid.org/0000-0002-9232-6176
authors.initials: R
authors.lastname: Urade
mesh_terms: D002384:Catalysis; D004220:Disulfides / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000201:enzymology; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D011490:Protein Disulfide Reductase (Glutathione) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics
chemical_list: D004220:Disulfides; D011994:Recombinant Proteins; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione); D019704:Protein Disulfide-Isomerases