Abstract
Oxidative protein folding in the endoplasmic reticulum is supported by efficient electron relays driven by enzymatic reactions centering on the ERO1-protein-disulfide isomerase (PDI) pathway. A controlled in vitro oxygen consumption assay was carried out to analyze the ERO1-PDI reaction. The results showed the pH-dependent oxidation of PDI by ERO1α. Among several possible disulfide bonds regulating ERO1α activity, Cys(94)-Cys(131) and Cys(99)-Cys(104) disulfide bonds are dominant regulators by excluding the involvement of the Cys(85)-Cys(391) disulfide in the regulation. The fine-tuned species specificity of the ERO1-PDI pathway was demonstrated by functional in vitro complementation assays using yeast and mammalian oxidoreductases. Finally, the results provide experimental evidence for the intramolecular electron transfer from the a domain to the a' domain within PDI during its oxidation by ERO1α.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cysteine / chemistry
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Cysteine / genetics
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Cysteine / metabolism
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Glycoproteins / chemistry
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Glycoproteins / genetics
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Glycoproteins / metabolism*
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Humans
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Membrane Glycoproteins / chemistry
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism*
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Oxidation-Reduction
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Oxidoreductases / chemistry
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Oxidoreductases / genetics
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Oxidoreductases / metabolism*
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Oxidoreductases Acting on Sulfur Group Donors / chemistry
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Oxidoreductases Acting on Sulfur Group Donors / genetics
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Oxidoreductases Acting on Sulfur Group Donors / metabolism*
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Protein Disulfide-Isomerases / chemistry
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Protein Disulfide-Isomerases / genetics
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Protein Disulfide-Isomerases / metabolism*
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Protein Structure, Tertiary
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae Proteins / chemistry
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism*
Substances
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Glycoproteins
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Membrane Glycoproteins
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Saccharomyces cerevisiae Proteins
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ERO1A protein, human
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Oxidoreductases
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Oxidoreductases Acting on Sulfur Group Donors
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ERO1 protein, S cerevisiae
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Protein Disulfide-Isomerases
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Cysteine