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Cysteine reactivity profiling to unveil redox regulation in phytopathogens

Abstract:
Reactivity-based chemical proteomics is a powerful technology based on the use of tagged chemicals that covalently react with surface-exposed residues on proteins in native proteomes. Reactivity profiling involves the purification, identification, and quantification of labeled peptides by LC-MS/MS. Here, we have detailed a protocol for reactivity profiling of Cys residues using iodoacetamide probes, displaying >1000 reactive Cys residues in the proteome of phytopathogen Pseudomonas syringae pv. tomato DC3000 (PtoDC3000). Comparative reactivity profiling of PtoDC3000 treated with or without hydrogen peroxide (H2O2) identified ~200 H2O2-sensitive Cys residues in antioxidant enzymes, metabolic enzymes, and transcription regulators. Interestingly, half of these H2O2-sensitive Cys residues are more reactive in response to H2O2 and several proteins have multiple Cys residues with opposite reactivities in response to H2O2 exposure.
Publication status:
Published
Peer review status:
Peer reviewed

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Files:
Publisher copy:
10.1007/978-1-0716-2079-3_9

Authors


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Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Sub department:
Plant Sciences
Research group:
Plant Chemetics Laboratory
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MSD
Department:
Biochemistry
Research group:
Advanced Proteomics Centre
Role:
Author
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Chemistry
Sub department:
Physical & Theoretical Chem
Role:
Author
ORCID:
0000-0003-2640-9560
More by this author
Institution:
University of Oxford
Division:
MPLS
Department:
Biology
Sub department:
Plant Sciences
Research group:
Plant Chemetics Laboratory
Oxford college:
Somerville College
Role:
Author
ORCID:
0000-0002-3692-7487


More from this funder
Funder identifier:
https://ror.org/0439y7842
Grant:
EP/X527245/1


Publisher:
Springer Nature
Host title:
Plant Proteases and Plant Cell Death: Methods and Protocols
Pages:
105-117
Series:
Methods in Molecular Biology
Series number:
2447
Place of publication:
New York, NY, USA
Publication date:
2022-05-19
DOI:
EISSN:
1940-6029
ISSN:
1064-3745
Pmid:
35583776
EISBN:
9781071620793
ISBN:
9781071620786


Language:
English
Keywords:
Subtype:
Chapter
Pubs id:
1261813
Local pid:
pubs:1261813
Deposit date:
2025-04-29

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