FRET reveals multiple interaction states between two component signalling system proteins of M. tuberculosis
FRET reveals multiple interaction states between two component signalling system proteins of M. tuberculosis
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DOI：
10.1016/j.bbagen.2016.04.011
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发表时间：
2016-07-01
影响因子：
3
通讯作者：
Saini, Deepak Kumar
中科院分区：
文献类型：
--
作者：
Agrawal, Ruchi;Kumar, Prem, V;Saini, Deepak Kumar
Background: Two component signalling involves interaction between sensor kinase (SIC) 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 SIC and RR, which was reflected as reduction in FRET ratio.General significance: 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. (C) 2016 Elsevier B.V. All rights reserved.
