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Measurement of mixed disulfides including glutathionylated proteins
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posted on 2023-06-07, 15:34 authored by Raffaella Priora, Lucia Coppo, Sonia Salzano, Paolo Di Simplicio, Pietro GhezziMixed disulfides between protein cysteines and low-molecular-weight thiol cysteine or glutathione lead to the formation of cysteinylated proteins or glutathionylated proteins. These types of posttranslational modification are of great importance in the so-called redox regulation, by which changes in the redox state of the cell regulate a number of biochemical processes. We describe the methods for quantitatively measuring the various redox states of cellular thiols including protein cysteines and these mixed disulfides. These include spectrophotometric methods, which do not distinguish between protein-cysteine and protein-glutathione disulfides, and HPLC methods that make such distinction. Finally, we report a method for labeling proteins susceptible to glutathionylation with biotin, to allow their visualization by Western blot after electrophoretic separation, which is used to identify proteins undergoing this posttranslational modification.
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Publication status
- Published
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- Published version
Journal
Methods in EnzymologyPublisher
Elsevier/Academic PressExternal DOI
Volume
473Page range
149-159Pages
359.0Book title
Thiol redox transitions in cell signaling. Part A, Chemistry and biochemistry of low molecular weight and protein thiolsPlace of publication
Amsterdam; BostonISBN
9780123813459Series
Methods in enzymologyDepartment affiliated with
- Clinical and Experimental Medicine Publications
Full text available
- No
Peer reviewed?
- Yes
Editors
Lester Packer, Enrique CadenasLegacy Posted Date
2010-10-25First Compliant Deposit (FCD) Date
2017-12-01Usage metrics
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