FRET reveals multiple interaction states between two component signalling system proteins of M. tuberculosis.
Biochim Biophys Acta
; 1860(7): 1498-507, 2016 Jul.
Article
em En
| MEDLINE
| ID: mdl-27102281
ABSTRACT
BACKGROUND:
Two component signalling involves interaction between sensor kinase (SK) and response regulator (RR) proteins which depends on their phosphorylation status.METHODS:
In this study we report the development of an in vitro FRET assay for studying interaction between fluorescently tagged SK and RR proteins.RESULTS:
Using TCS proteins of Mycobacterium tuberculosis, we demonstrate that phosphorylation status of SK affects the SK-RR interaction, which varies from one TCS to another. The observation was strengthened by recordings from mutant SK and RR proteins. The assay retained the specificity/crosstalk potential of the participating proteins and reflected the inherent phosphotransfer potentials.CONCLUSIONS:
SK and RR proteins interact with each other in unphosphorylated state and the phosphorylation affects the interaction between SK and RR, which was reflected as reduction in FRET ratio. GENERALSIGNIFICANCE:
A non-radioactive, in vitro FRET based assay is reported, which can be utilized for studying genome-wide partner screening, identifying crosstalk or specificity in TCSs.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas Quinases
/
Proteínas de Bactérias
/
Técnicas Biossensoriais
/
Transdução de Sinais
/
Transativadores
/
Transferência Ressonante de Energia de Fluorescência
/
Mycobacterium tuberculosis
Tipo de estudo:
Prognostic_studies
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article