The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation.
Glutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2006
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| _version_ | 1826261574411616256 |
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| author | Brennan, J Miller, J Fuller, W Wait, R Begum, S Dunn, M Eaton, P |
| author_facet | Brennan, J Miller, J Fuller, W Wait, R Begum, S Dunn, M Eaton, P |
| author_sort | Brennan, J |
| collection | OXFORD |
| description | Glutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or regulate protein function. We developed novel strategies for the study of protein S-glutathiolation that involved the simple synthesis of N,N-biotinyl glutathione disulfide (biotin-GSSG). Biotin-GSSG treatment of cells mimics a defined component of oxidative stress, namely a shift in the glutathione redox couple to the oxidized disulfide state. This induces widespread protein S-glutathiolation, which was detected on non-reducing Western blots probed with streptavidin-horseradish peroxidase and imaged using confocal fluorescence microscopy and ExtrAvidin-FITC. S-Glutathiolated proteins were purified using streptavidin-agarose and identified using proteomic methods. We conclude that biotin-GSSG is a useful tool in the investigation of protein S-glutathiolation and offers significant advantages over conventional methods or antibody-based strategies. These novel approaches may find widespread utility in the study of disease or redox signaling models where GSSG accumulation occurs. |
| first_indexed | 2024-03-06T19:23:30Z |
| format | Journal article |
| id | oxford-uuid:1aebfcef-9a1e-4f04-bffa-fdfa884992fd |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T19:23:30Z |
| publishDate | 2006 |
| record_format | dspace |
| spelling | oxford-uuid:1aebfcef-9a1e-4f04-bffa-fdfa884992fd2022-03-26T10:57:26ZThe utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:1aebfcef-9a1e-4f04-bffa-fdfa884992fdEnglishSymplectic Elements at Oxford2006Brennan, JMiller, JFuller, WWait, RBegum, SDunn, MEaton, PGlutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or regulate protein function. We developed novel strategies for the study of protein S-glutathiolation that involved the simple synthesis of N,N-biotinyl glutathione disulfide (biotin-GSSG). Biotin-GSSG treatment of cells mimics a defined component of oxidative stress, namely a shift in the glutathione redox couple to the oxidized disulfide state. This induces widespread protein S-glutathiolation, which was detected on non-reducing Western blots probed with streptavidin-horseradish peroxidase and imaged using confocal fluorescence microscopy and ExtrAvidin-FITC. S-Glutathiolated proteins were purified using streptavidin-agarose and identified using proteomic methods. We conclude that biotin-GSSG is a useful tool in the investigation of protein S-glutathiolation and offers significant advantages over conventional methods or antibody-based strategies. These novel approaches may find widespread utility in the study of disease or redox signaling models where GSSG accumulation occurs. |
| spellingShingle | Brennan, J Miller, J Fuller, W Wait, R Begum, S Dunn, M Eaton, P The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title | The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title_full | The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title_fullStr | The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title_full_unstemmed | The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title_short | The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. |
| title_sort | utility of n n biotinyl glutathione disulfide in the study of protein s glutathiolation |
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