FRET reveals multiple interaction states between two component signalling system proteins of M. tuberculosis

Agrawal, Ruchi and Kumar, Prem and Ramanan, Harini and Saini, Deepak Kumar (2016) FRET reveals multiple interaction states between two component signalling system proteins of M. tuberculosis. In: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 1860 (7). pp. 1498-1507.

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Abstract

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.

Item Type:	Journal Article
Publication:	BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
Publisher:	ELSEVIER SCIENCE BV
Additional Information:	Copy right for this article belongs to the ELSEVIER SCIENCE BV, PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS
Keywords:	Two component signalling; Sensor kinase; Response regulator; Crosstalk; FRET
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics Division of Interdisciplinary Sciences > Centre for Biosystems Science and Engineering
Date Deposited:	30 Jun 2016 05:39
Last Modified:	11 Oct 2018 07:27
URI:	http://eprints.iisc.ac.in/id/eprint/54076

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