title: Identification and characterization of GmPDIL7, a soybean ER membrane-bound protein disulfide isomerase family protein.
abstract: Most proteins synthesized in the endoplasmic reticulum (ER) possess intramolecular and intermolecular disulfide bonds, which play an important role in the conformational stability and function of proteins. Hence, eukaryotic cells contain protein disulfide bond formation pathways such as the protein disulfide isomerase (PDI)-ER oxidoreductin 1 (Ero1) system in the ER lumen. In this study, we identified soybean PDIL7 (GmPDIL7), a novel soybean ER membrane-bound PDI family protein, and determined its enzymatic properties. GmPDIL7 has a putative N-terminal signal sequence, a thioredoxin domain with an active center motif (CGHC), and a putative C-terminal transmembrane region. Likewise, we demonstrated that GmPDIL7 is ubiquitously expressed in soybean tissues and is localized in the ER membrane. Furthermore, GmPDIL7 associated with other soybean PDI family proteins in vivo and GmPDIL7 mRNA was slightly upregulated under ER stress. The redox potential of recombinant GmPDIL7 expressed in Escherichia coli was -187 mV, indicating that GmPDIL7 could oxidize unfolded proteins. GmPDIL7 exhibited a dithiol oxidase activity level that was similar to other soybean PDI family proteins. However, the oxidative refolding activity of GmPDIL7 was lower than other soybean PDI family proteins. GmPDIL7 was well oxidized by GmERO1. Taken together, our results indicated that GmPDIL7 primarily plays a role as a supplier of disulfide bonds in nascent proteins for oxidative folding on the ER membrane.
  
  
  
  The nucleotide sequence data for the GmPDIL7 cDNA are available in the DNA Data Bank of Japan (DDBJ) databases under the accession numbers LC158001.
  
  
  
  Protein disulfide isomerase: EC 5.3.4.1.
authors.affiliation: Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Uji, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Okuda
authors.affiliation: Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Uji, Japan.
authors.forename: Motonori
authors.initials: M
authors.lastname: Matsusaki
authors.affiliation: Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Uji, Japan.
authors.forename: Taro
authors.initials: T
authors.lastname: Masuda
authors.affiliation: Division of Agronomy and Horticultural Science, Graduate School of Agriculture, Kyoto University, Uji, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Urade
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000072417:Protein Domains; D021382:Protein Sorting Signals; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013025:Glycine max / Q000737:chemistry* / Q000201:enzymology
chemical_list: D010940:Plant Proteins; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019704:Protein Disulfide-Isomerases
keywords: ERO1; endoplasmic reticulum; protein disulfide isomerase; protein folding; soybean