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Structure of the SAS-6 cartwheel hub from Leishmania major

DOI: 10.7554/eLife.01812 DOI Help
PMID: 24596152 PMID Help

Authors: Mark Van Breugel (MRC Laboratory of Molecular Biology) , Rainer Wilcken (MRC Centre) , Stephen H. Mclaughlin (MRC Laboratory of Molecular Biology) , Trevor J. Rutherford (MRC Laboratory of Molecular Biology) , Christopher M. Johnson (MRC Laboratory of Molecular Biology)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Elife , VOL 3 , PAGES e01812 - e01812

State: Published (Approved)
Published: January 2014
Diamond Proposal Number(s): 8547

Open Access Open Access

Abstract: The covalent attachment of adenosine monophosphate (AMP) to proteins, a process called AMPylation (adenylylation), has recently emerged as a novel theme in microbial pathogenesis. Although several AMPylating enzymes have been characterized, the only known virulence protein with de-AMPylation activity is SidD from the human pathogen Legionella pneumophila. SidD de-AMPylates mammalian Rab1, a small GTPase involved in secretory vesicle transport, thereby targeting the host protein for inactivation. The molecular mechanisms underlying Rab1 recognition and de-AMPylation by SidD are unclear. Here, we report the crystal structure of the catalytic region of SidD at 1.6 Å resolution. The structure reveals a phosphatase-like fold with additional structural elements not present in generic PP2C-type phosphatases. The catalytic pocket contains a binuclear metal-binding site characteristic of hydrolytic metalloenzymes, with strong dependency on magnesium ions. Subsequent docking and molecular dynamics simulations between SidD and Rab1 revealed the interface contacts and the energetic contribution of key residues to the interaction. In conjunction with an extensive structure-based mutational analysis, we provide in vivo and in vitro evidence for a remarkable adaptation of SidD to its host cell target Rab1 which explains how this effector confers specificity to the reaction it catalyses.

Subject Areas: Biology and Bio-materials

Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography

Other Facilities: ESRF ID29 and BM14

Added On: 20/03/2014 12:45

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