Molecular bases of cyclic and specific disulfide interchange between human ERO1α protein and protein-disulfide isomerase (PDI)

Shoji Masui, Stefano Vavassori, Claudio Fagioli, Roberto Sitia, Kenji Inaba

Research output: Contribution to journalArticle

Abstract

In the endoplasmic reticulum (ER) of human cells, ERO1α and protein-disulfide isomerase (PDI) constitute one of the major electron flow pathways that catalyze oxidative folding of secretory proteins. Specific and limited PDI oxidation by ERO1α is essential to avoid ER hyperoxidation. To investigate how ERO1α oxidizes PDI selectively among more than 20 ER-resident PDI family member proteins, we performed docking simulations and systematic biochemical analyses. Our findings reveal that a protruding β-hairpin of ERO1α specifically interacts with the hydrophobic pocket present in the redox-inactive PDI b′-domain through the stacks between their aromatic residues, leading to preferred oxidation of the C-terminal PDI a′-domain. ERO1α associated preferentially with reduced PDI, explaining the stepwise disulfide shuttle mechanism, first from ERO1α to PDI and then from oxidized PDI to an unfolded polypeptide bound to its hydrophobic pocket. The interaction of ERO1α with ERp44, another PDI family member protein, was also analyzed. Notably, ERO1α-dependent PDI oxidation was inhibited by a hyperactive ERp44 mutant that lacks the C-terminal tail concealing the substrate-binding hydrophobic regions. The potential ability of ERp44 to inhibit ERO1α activity may suggest its physiological role in ER redox and protein homeostasis.

Original languageEnglish
Pages (from-to)16261-16271
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number18
DOIs
Publication statusPublished - May 6 2011

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Protein Disulfide-Isomerases
Interchanges
Disulfides
Proteins
Endoplasmic Reticulum
Oxidation
Oxidation-Reduction
Aptitude
Protein Folding
Tail
Homeostasis

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Molecular bases of cyclic and specific disulfide interchange between human ERO1α protein and protein-disulfide isomerase (PDI). / Masui, Shoji; Vavassori, Stefano; Fagioli, Claudio; Sitia, Roberto; Inaba, Kenji.

In: Journal of Biological Chemistry, Vol. 286, No. 18, 06.05.2011, p. 16261-16271.

Research output: Contribution to journalArticle

Masui, Shoji ; Vavassori, Stefano ; Fagioli, Claudio ; Sitia, Roberto ; Inaba, Kenji. / Molecular bases of cyclic and specific disulfide interchange between human ERO1α protein and protein-disulfide isomerase (PDI). In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 18. pp. 16261-16271.
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