Detection of Disulfides in Protein Extracts of
Arabidopsis thaliana
Disulfide
Monobromobimane (mBB)
Protein labeling
Redox
Thiol
Tris(2-carboxyethyl) phosphine (TCEP)
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Mar 2019
05 Mar 2019
Historique:
received:
27
12
2018
revised:
28
02
2019
accepted:
12
02
2019
entrez:
3
3
2021
pubmed:
5
3
2019
medline:
5
3
2019
Statut:
epublish
Résumé
Thiol-disulfide exchange is a key posttranslational modification, determining the folding process of intra- and inter-protein structures. Thiols can be detected by colorimetric reagents, which are stoichiometrically reduced by free thiols, and by fluorescent adducts, showing fluorescence only after thioester formation. We adapted a simple three-step method for detection of disulfide bonds in proteins. After irreversible blocking of protein thiols, disulfide bonds are reduced, followed by the detection of thiols. The approach presented here provides an economical procedure that can be used to obtain a global overview of the thiol-disulfide status of proteins in plants. This method allows the detection of modifications in samples on a gel and can be used for semi-quantitative analysis.
Identifiants
pubmed: 33654985
doi: 10.21769/BioProtoc.3183
pii: e3183
pmc: PMC7854131
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e3183Informations de copyright
Copyright © 2019 The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsThe authors declare that there are no conflicts of interest.
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