DOI: 10.1002/pro.5560020503. Redox properties of protein disulfide isomerase (dsba) from <i>escherichia coli</i>

Redox properties of protein disulfide isomerase (dsba) from escherichia coli

10.1002/pro.5560020503

Crossref journal-article

Wiley

Protein Science (311)

Abstract

AbstractThe redox properties of periplasmic protein disulfide isomerase (DsbA) from Escherichia coli were analyzed by measuring the equilibrium constant of the oxidation of reduced DsbA by oxidized glutathione. The experiments are based on the finding that the intrinsic tryptophan fluorescence of DsbA increases about threefold upon reduction of the enzyme, which can be explained by the catalytic disulfide bridge quenching the fluorescence of a neighboring tryptophan residue. From the specific fluorescence of DsbA equilibrated in the presence of different ratios of reduced and oxidized glutathione at pH 7, an equilibrium constant of 1.2 × 10−4 M was determined, corresponding to a standard redox potential (E'0) of DsbA of ‐0.089 V. Thus, DsbA is a significantly stronger oxidant than cytoplasmic thioredoxins and its redox properties are similar to those of eukaryotic protein disulfide isomerase. The equilibrium constants for the DsbA/glutathione equilibrium were found to be strongly dependent on pH and varied from 2.5 × 10−3 M to 3.9 × 10−5 M between pH 4 and 8.5. The redox state‐dependent fluorescence properties of DsbA should allow detailed physicochemical studies of the enzyme as well as the quantitative determination of the oxidized protein by fluorescence titration with dithiothreitol and open the possibility to observe bacterial protein disulfide isomerase “at work” during catalysis of oxidative protein folding.

Bibliography

Wunderlich, M., & Glockshuber, R. (1993). Redox properties of protein disulfide isomerase (dsba) from escherichia coli. Protein Science, 2(5), 717â726. Portico.

Authors 2

Martina Wunderlich (first)
Rudi Glockshuber (additional)

References 34 Referenced 206

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{'key': 'e_1_2_1_26_2', 'first-page': '3553', 'article-title': 'Protein disulfide isomerase', 'volume': '267', 'author': 'Noiva R.', 'year': '1992', 'journal-title': 'J. Biol. Chem.'} / J. Biol. Chem. / Protein disulfide isomerase by Noiva R. (1992)
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Dates

Type	When
Created	16 years, 6 months ago (Feb. 9, 2009, 7:35 p.m.)
Deposited	1 year, 10 months ago (Oct. 24, 2023, 7:03 p.m.)
Indexed	2 months, 1 week ago (June 20, 2025, 1:26 p.m.)
Issued	32 years, 4 months ago (May 1, 1993)
Published	32 years, 4 months ago (May 1, 1993)
Published Online	16 years, 8 months ago (Dec. 31, 2008)
Published Print	32 years, 4 months ago (May 1, 1993)

Funders 0

None

BibTeX

@article{Wunderlich_1993, title={Redox properties of protein disulfide isomerase (dsba) from escherichia coli}, volume={2}, ISSN={1469-896X}, url={http://dx.doi.org/10.1002/pro.5560020503}, DOI={10.1002/pro.5560020503}, number={5}, journal={Protein Science}, publisher={Wiley}, author={Wunderlich, Martina and Glockshuber, Rudi}, year={1993}, month=may, pages={717–726} }

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  "abstract": "<jats:title>Abstract</jats:title><jats:p>The redox properties of periplasmic protein disulfide isomerase (DsbA) from <jats:italic>Escherichia coli</jats:italic> were analyzed by measuring the equilibrium constant of the oxidation of reduced DsbA by oxidized glutathione. The experiments are based on the finding that the intrinsic tryptophan fluorescence of DsbA increases about threefold upon reduction of the enzyme, which can be explained by the catalytic disulfide bridge quenching the fluorescence of a neighboring tryptophan residue. From the specific fluorescence of DsbA equilibrated in the presence of different ratios of reduced and oxidized glutathione at pH 7, an equilibrium constant of 1.2 \u00d7 10<jats:sup>\u22124</jats:sup> M was determined, corresponding to a standard redox potential (<jats:italic>E</jats:italic>'<jats:sub>0</jats:sub>) of DsbA of \u20100.089 V. Thus, DsbA is a significantly stronger oxidant than cytoplasmic thioredoxins and its redox properties are similar to those of eukaryotic protein disulfide isomerase. The equilibrium constants for the DsbA/glutathione equilibrium were found to be strongly dependent on pH and varied from 2.5 \u00d7 10<jats:sup>\u22123</jats:sup> M to 3.9 \u00d7 10<jats:sup>\u22125</jats:sup> M between pH 4 and 8.5. The redox state\u2010dependent fluorescence properties of DsbA should allow detailed physicochemical studies of the enzyme as well as the quantitative determination of the oxidized protein by fluorescence titration with dithiothreitol and open the possibility to observe bacterial protein disulfide isomerase \u201cat work\u201d during catalysis of oxidative protein folding.</jats:p>",
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