title: Glutathione peroxidase 7 utilizes hydrogen peroxide generated by Ero1α to promote oxidative protein folding.
abstract: OBJECTIVE
  Ero1 flavoproteins catalyze oxidative folding in the endoplasmic reticulum (ER), consuming oxygen and generating hydrogen peroxide (H2O2). The ER-localized glutathione peroxidase 7 (GPx7) shows protein disulfide isomerase (PDI)-dependent peroxidase activity in vitro. Our work aims at identifying the physiological role of GPx7 in the Ero1α/PDI oxidative folding pathway and at dissecting the reaction mechanisms of GPx7.
  
  
  RESULTS
  Our data show that GPx7 can utilize Ero1α-produced H2O2 to accelerate oxidative folding of substrates both in vitro and in vivo. H2O2 oxidizes Cys57 of GPx7 to sulfenic acid, which can be resolved by Cys86 to form an intramolecular disulfide bond. Both the disulfide form and sulfenic acid form of GPx7 can oxidize PDI for catalyzing oxidative folding. GPx7 prefers to interact with the a domain of PDI, and intramolecular cooperation between the two redox-active sites of PDI increases the activity of the Ero1α/GPx7/PDI triad.
  
  
  METHODS
  Our in vitro and in vivo evidence provides mechanistic insights into how cells consume potentially harmful H2O2 while optimizing oxidative protein folding via the Ero1α/GPx7/PDI triad. Cys57 can promote PDI oxidation in two ways, and Cys86 emerges as a novel noncanonical resolving cysteine.
  
  
  CONCLUSIONS
  GPx7 promotes oxidative protein folding, directly utilizing Ero1α-generated H2O2 in the early secretory compartment. Thus, the Ero1α/GPx7/PDI triad generates two disulfide bonds and two H2O molecules at the expense of a single O2 molecule.
authors.affiliation: 1 National Laboratory of Biomacromolecules, Institute of Biophysics , Chinese Academy of Sciences, Beijing, China .
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Lihui
authors.initials: L
authors.lastname: Zhang
authors.forename: Yingbo
authors.initials: Y
authors.lastname: Niu
authors.forename: Roberto
authors.initials: R
authors.lastname: Sitia
authors.forename: Chih-Chen
authors.initials: CC
authors.lastname: Wang
mesh_terms: D020134:Catalytic Domain; D005979:Glutathione Peroxidase; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry* / Q000378:metabolism; D007144:Immunoglobulin J-Chains / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry* / Q000378:metabolism; D010544:Peroxidases / Q000737:chemistry* / Q000502:physiology; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000737:chemistry; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry
chemical_list: D007144:Immunoglobulin J-Chains; D008562:Membrane Glycoproteins; D006861:Hydrogen Peroxide; C405327:ERO1A protein, human; D010088:Oxidoreductases; D010544:Peroxidases; C558641:GPX7 protein, human; D005979:Glutathione Peroxidase; D012259:Ribonuclease, Pancreatic; D019704:Protein Disulfide-Isomerases
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