PXD048374

PXD048374 is an original dataset announced via ProteomeXchange.

Title	Specificity and dynamics of H2O2 detoxification by the cytosolic redox regulatory network as revealed by in vitro reconstitution
Description	The thiol redox state is a decisive functional characteristic of proteins in cell biology. Plasmatic cell compartments maintain a thiol-based redox regulatory network linked to the glutathione/glutathione disulfide couple (GSH/GSSG) and the NAD(P)H system. The basic network constituents are known and in vivo cell imaging with gene-encoded probes have revealed insight into the dynamics of the [GSH]2/[GSSG] redox potential, H2O2 and NAD(P)H+H+ in dependence on the metabolic and environmental conditions. Highly limited is our understanding of the contribution and interaction of the components in the network, also because of compensatory reactions in genetic approaches. Reconstituting the cytosolic network in vitro from fifteen recombinant proteins at in vivo concentrations, namely glutathione peroxidase-like (GPXL), peroxiredoxins (PRX), glutaredoxins (GRX), thioredoxins (TRX), NADPH-dependent thioredoxin reductase A (NTRA) and glutathione reductase (GR) and applying GRX1-roGFP2 or roGFP2-ORP1 as dynamic sensors, allows for monitoring the response to a single H2O2 pulse. The major change in thiol oxidation as visualized by targeted proteomics occurred in relevant peptides of GPXL, and to a lesser extent of PRX, while other Cys-containing peptides only showed small changes in redox state and protection. Titration of ascorbate peroxidase (APX2) into the system together with dehydroascorbate reductase (DHAR1) lowered the oxidation of the fluorescent sensors in the network, but was unable to suppress it. The results demonstrate the power of the network to detoxify H2O2, the partially independent branches of electron flow with significance for specific cell signaling and the importance of APX to modulate the signaling without suppressing it and shifting the burden to glutathione oxidation.
HostingRepository	jPOST
AnnounceDate	2025-01-21
AnnouncementXML	Submission_2025-01-21_04:59:31.838.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jürgen Eirich
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	NEM:2H(5); alpha-amino acetylated residue; N-ethylmaleimide derivatized cysteine; L-methionine sulfoxide
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-01-10 00:27:31	ID requested
⏵ 1	2025-01-21 04:59:32	announced

Vogelsang L, Eirich J, Finkemeier I, Dietz KJ, detoxification by the cytosolic redox regulatory network as revealed by in vitro reconstitution. Redox Biol, 72():103141(2024) [pubmed]

submitter keyword: Glutathione peroxidase, hydrogen peroxide, peroxiredoxin, reactive oxygen species, thioredoxin, isotope labeling

Iris Finkemeier
lab head
Jürgen Eirich
contact affiliation	University of Münster - WWU - Institute for Biology and Biotechnology of Plants
dataset submitter

jPOST dataset URI

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