Proteome-wide tagging with an H 2 O 2 biosensor reveals highly localized and dynamic redox microenvironments
Hydrogen peroxide (H 2 O 2 ) sensing and signaling involves the reversible oxidation of particular thiols on particular proteins to modulate protein function in a dynamic manner. H 2 O 2 can be generated from various intracellular sources, but their identities and relative contributions are often un...
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| Published in | Proceedings of the National Academy of Sciences - PNAS Vol. 120; no. 48 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
28.11.2023
|
| Online Access | Get full text |
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| Summary: | Hydrogen peroxide (H
2
O
2
) sensing and signaling involves the reversible oxidation of particular thiols on particular proteins to modulate protein function in a dynamic manner. H
2
O
2
can be generated from various intracellular sources, but their identities and relative contributions are often unknown. To identify endogenous “hotspots” of H
2
O
2
generation on the scale of individual proteins and protein complexes, we generated a yeast library in which the H
2
O
2
sensor HyPer7 was fused to the C-terminus of all protein-coding open reading frames (ORFs). We also generated a control library in which a redox-insensitive mutant of HyPer7 (SypHer7) was fused to all ORFs. Both libraries were screened side-by-side to identify proteins located within H
2
O
2
-generating environments. Screening under a variety of different metabolic conditions revealed dynamic changes in H
2
O
2
availability highly specific to individual proteins and protein complexes. These findings suggest that intracellular H
2
O
2
generation is much more localized and functionally differentiated than previously recognized. |
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| ISSN: | 0027-8424 1091-6490 |
| DOI: | 10.1073/pnas.2314043120 |