Determination of glutathione-binding to proteins by fluorescence spectroscopy.
Biochem Biophys Res Commun
; 557: 329-333, 2021 06 11.
Article
em En
| MEDLINE
| ID: mdl-33895474
ABSTRACT
Glutathione (GSH) is the most abundant non-protein thiol and its cellular concentration has been reported as 17 mM in Escherichia coli. This study introduces a label-free method to determine the binding affinity of GSH to proteins, utilizing the intrinsic fluorescence of proteins; the dissociation constants of GSH for d-arabinose 5-phosphate isomerase KdsD, fumarase C, malate dehydrogenase, and RNA polymerase subunit α have been determined as 96 ± 8, 246 ± 42, 292 ± 78, and 296 ± 97 µM, respectively. The dissociation constants, less than 2% of the cellular concentration of GSH, suggests that protein-GSH interactions are strong enough to make all of the GSH-binding sites occupied fully. The method described here may be applicable to other proteins.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Fluorescência
/
RNA Polimerases Dirigidas por DNA
/
Aldose-Cetose Isomerases
/
Fumarato Hidratase
/
Glutationa
/
Malato Desidrogenase
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article