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Functional Disulphide Bonds pp 133–148Cite as

Assays of Thiol Isomerase Enzymatic Activity

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1967))

Abstract

Thiol isomerases are oxidoreductases that mediate disulphide bond formation in nascent proteins of the endoplasmic reticulum to ensure their structural integrity. In addition to its role in protein folding, thiol isomerases can modify allosteric disulphide bonds in both intracellular and extracellular proteins, thereby controlling protein function. The process of disulphide bond formation and cleavage is strictly regulated and responsive to redox conditions. Understanding disulphide bond regulation under different redox environments is critical to understanding physiological and pathological processes related to disulphide bond chemistry. Here we describe protocols for the measurement of disulphide bond modulation by thiol isomerases, including reductase and denitrosylase assays. These methods can be applied to study recombinant thiol isomerases and thiol isomerases in cellular settings.

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Authors and Affiliations

  1. Division of Hemostasis and Thrombosis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Roelof H. Bekendam & Robert Flaumenhaft

Authors
  1. Roelof H. Bekendam
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  2. Robert Flaumenhaft
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Corresponding author

Correspondence to Robert Flaumenhaft .

Editor information

Editors and Affiliations

  1. The Centenary Institute, NHMRC Clinical Trials Centre, Sydney Medical School, University of Sydney, Sydney, NSW, Australia

    Philip Hogg

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Bekendam, R.H., Flaumenhaft, R. (2019). Assays of Thiol Isomerase Enzymatic Activity. In: Hogg, P. (eds) Functional Disulphide Bonds. Methods in Molecular Biology, vol 1967. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9187-7_8

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  • DOI: https://doi.org/10.1007/978-1-4939-9187-7_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9186-0

  • Online ISBN: 978-1-4939-9187-7

  • eBook Packages: Springer Protocols

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