Structural Insight into the Mechanism of C-di-GMP Hydrolysis by EAL Domain Phosphodiesterases

Overview

Journal J Mol Biol

Publisher Elsevier

Specialties Microbiology
Molecular Biology

Date 2010 Aug 10

PMID 20691189

Citations 66

Authors

Anatoli Tchigvintsev

Xiaohui Xu

Alexander Singer

Changsoo Chang

Greg Brown

Michael Proudfoot

Hong Cui

Robert Flick

Wayne F Anderson

Andrzej Joachimiak

Michael Y Galperin

Alexei Savchenko

Alexander F Yakunin

Affiliations

Soon will be listed here.

Abstract

Cyclic diguanylate (or bis-(3'-5') cyclic dimeric guanosine monophosphate; c-di-GMP) is a ubiquitous second messenger that regulates diverse cellular functions, including motility, biofilm formation, cell cycle progression, and virulence in bacteria. In the cell, degradation of c-di-GMP is catalyzed by highly specific EAL domain phosphodiesterases whose catalytic mechanism is still unclear. Here, we purified 13 EAL domain proteins from various organisms and demonstrated that their catalytic activity is associated with the presence of 10 conserved EAL domain residues. The crystal structure of the TBD1265 EAL domain was determined in free state (1.8 Å) and in complex with c-di-GMP (2.35 A), and unveiled the role of conserved residues in substrate binding and catalysis. The structure revealed the presence of two metal ions directly coordinated by six conserved residues, two oxygens of c-di-GMP phosphate, and potential catalytic water molecule. Our results support a two-metal-ion catalytic mechanism of c-di-GMP hydrolysis by EAL domain phosphodiesterases.

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