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000294676 245__ $$aUsing the heme peroxidase APEX2 to probe intracellular H2O2 flux and diffusion.
000294676 260__ $$a[London]$$bNature Publishing Group UK$$c2024
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000294676 520__ $$aCurrently available genetically encoded H2O2 probes report on the thiol redox state of the probe, which means that they reflect the balance between probe thiol oxidation and reduction. Here we introduce the use of the engineered heme peroxidase APEX2 as a thiol-independent chemogenetic H2O2 probe that directly and irreversibly converts H2O2 molecules into either fluorescent or luminescent signals. We demonstrate sensitivity, specificity, and the ability to quantitate endogenous H2O2 turnover. We show how the probe can be used to detect changes in endogenous H2O2 generation and to assess the roles and relative contributions of endogenous H2O2 scavengers. Furthermore, APEX2 can be used to study H2O2 diffusion inside the cytosol. Finally, APEX2 reveals the impact of commonly used alkylating agents and cell lysis protocols on cellular H2O2 generation.
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000294676 536__ $$0G:(EU-Grant)742039$$aRedox Relays - Detecting, understanding and exploiting intracellular redox signaling relays (742039)$$c742039$$fERC-2016-ADG$$x1
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000294676 650_7 $$042VZT0U6YR$$2NLM Chemicals$$aHeme
000294676 650_7 $$0BBX060AN9V$$2NLM Chemicals$$aHydrogen Peroxide
000294676 650_7 $$0EC 1.11.1.-$$2NLM Chemicals$$aPeroxidases
000294676 650_7 $$2NLM Chemicals$$aSulfhydryl Compounds
000294676 650_2 $$2MeSH$$aHeme
000294676 650_2 $$2MeSH$$aHydrogen Peroxide
000294676 650_2 $$2MeSH$$aOxidation-Reduction
000294676 650_2 $$2MeSH$$aPeroxidases: chemistry
000294676 650_2 $$2MeSH$$aPeroxidases: metabolism
000294676 650_2 $$2MeSH$$aSulfhydryl Compounds
000294676 7001_ $$0P:(DE-He78)387d4f13f0c0f8a92fee635038e7d424$$aBarayeu, Uladzimir$$b1
000294676 7001_ $$0P:(DE-He78)86ad31c8d73f6fda34fc756afb17dff8$$aSulková, Katerina$$b2$$udkfz
000294676 7001_ $$0P:(DE-HGF)0$$aAranda-Vallejo, Carla$$b3
000294676 7001_ $$0P:(DE-He78)7f55a0ed8b021080de00960cc73768fb$$aDick, Tobias$$b4$$eLast author$$udkfz
000294676 773__ $$0PERI:(DE-600)2553671-0$$a10.1038/s41467-024-45511-9$$gVol. 15, no. 1, p. 1239$$n1$$p1239$$tNature Communications$$v15$$x2041-1723$$y2024
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