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The crystal structure of the lipid II-degrading bacteriocin syringacin M suggests unexpected evolutionary relationships between colicin M-like bacteriocins

DOI: 10.1074/jbc.M112.400150 DOI Help
PMID: 22995910 PMID Help

Authors: Rhys Grinter (University of Glasgow) , A Roszak (University of Glasgow) , R. J. Cogdell (University of Glasgow) , J. J. Milner (University of Glasgow) , D. Walker (University of Glasgow)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of Biological Chemistry

State: Published (Approved)
Published: September 2012
Diamond Proposal Number(s): 6683

Abstract: Colicin-like bacteriocins show potential as next generation antibiotics with clinical and agricultural applications. Key to these potential applications is their high potency and species specificity that enables a single pathogenic species to be targeted with minimal disturbance of the wider microbial community. Here we present the structure and function of the colicin M-like bacteriocin, syringacin M from Pseudomonas syringae pv. tomato DC3000. Syringacin M kills susceptible cells through a highly specific phosphatase activity that targets lipid II, ultimately inhibiting peptidoglycan synthesis. Comparison of the structures of syringacin M and colicin M reveals that, in addition to the expected similarity between the homologous C-terminal catalytic domains, the receptor binding domains of these proteins, which share no discernible sequence homology, share a striking structural similarity. This indicates that the generation of the novel receptor binding and species specificities of these bacteriocins has been driven by diversifying selection rather than diversifying recombination as suggested previously. Additionally, the structure of syringacin M reveals the presence of an active site calcium ion that is coordinated by a conserved aspartic acid side chain and is essential for catalytic activity. We show that mutation of this residue to alanine inactivates syringacin M and that the metal ion is absent from the structure of the mutant protein. Consistent with the presence of Ca2+ in the active site, we show that syringacin M activity is supported by Ca2+, along with Mg2+ and Mn2+, and the protein is catalytically inactive in the absence of these ions.

Journal Keywords: Calcium-binding Proteins; Crystal Structure; Metalloenzymes; Pseudomonas; X-ray Crystallography; Bacteriocin; Colicin M; Divergent Evolution; Lipid II; Syringacin M

Subject Areas: Biology and Bio-materials, Chemistry, Food Science


Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography

Other Facilities: No