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Synthesis and Characterization of a Fluorescent Analogue of Cyclic di-GMP

Sharma, Indra Mani and Dhanaraman, Thillaivillalan and Mathew, Ritta and Chatterji, Dipankar (2012) Synthesis and Characterization of a Fluorescent Analogue of Cyclic di-GMP. In: BIOCHEMISTRY, 51 (27). pp. 5443-5453.

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Official URL: http://dx.doi.org/10.1021/bi3003617

Abstract

Cyclic di-GMP (c-di-GMP), a ubiquitous bacterial second messenger, has emerged as a key controller of several biological processes. Numbers of reports that deal with the mechanistic aspects of this second messenger have appeared in the literature. However, the lack of a reporter tag attached to the c-di-GMP at times limits the understanding of further details. In this study, we have chemically coupled N-methylisatoic anhydride (MANT) with c-di-GMP, giving rise to Mant-(c-di-GMP) or MANT-CDG. We have characterized the chemical and physical properties and spectral behavior of MANT-CDG. The fluorescence of MANT-CDG is sensitive to changes in the microenvironment, which helped us study its interaction with three different c-di-GMP binding proteins (a diguanylate cyclase, a phosphodiesterase, and a PilZ domain-containing protein). In addition, we have shown here that MANT-CDG can inhibit diguanylate cyclase activity; however, it is hydrolyzed by c-di-GMP specific phosphodiesterase. Taken together, our data suggest that MANT-CDG behaves like native c-di-GMP, and this study raises the possibility that MANT-CDG will be a valuable research tool for the in vitro characterization of c-di-GMP signaling factors.

Item Type: Journal Article
Additional Information: Copyright for this article belongs to the American Chemical Society
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Date Deposited: 09 Aug 2012 11:26
Last Modified: 09 Aug 2012 11:27
URI: http://eprints.iisc.ernet.in/id/eprint/44918

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