3qay

Catalytic domain of CD27L endolysin targeting Clostridia Difficile

spinqualitylabelsall models 3qay, resolution 2.00Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 3qay is a 4 chain structure with sequence from Clostridium phage phicd27. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Ligands: PO4, ZN Resources: FirstGlance, OCA, RCSB, PDBsum Publication Abstract from PubMed Endolysin CD27L causes cell lysis of the pathogen Clostridium difficile, a major cause of nosocomial infection. We report a structural and functional analysis of the catalytic activity of CD27L against C. difficile and other bacterial strains. We show that truncation of the endolysin to the N-terminal domain, CD27L1-179, gave an increased lytic activity against cells of C. difficile, while the C-terminal region, CD27L180-270, failed to produce lysis. CD27L1-179 also has increased activity against other bacterial species that are targeted by the full-length protein and in addition was able to lyse some CD27L-insensitive strains. However, CD27L1-179 retained a measure of specificity, failing to lyse a wide range of bacteria. The use of green fluorescent protein (GFP)-labeled proteins demonstrated that both CD27L and CD27L1-179 bound to C. difficile cell walls. The crystal structure of CD27L1-179 confirms that the enzyme is a zinc-dependent N-acetylmuramoyl-l-alanine amidase. A structure-based sequence analysis allowed us to identify four catalytic residues, a proton relay cascade, and a substrate binding pocket. A BLAST search shows that the closest-related amidases almost exclusively target Clostridia. This implied that the catalytic domain alone contained features that target a specific bacterial species. To test this hypothesis, we modified Leu 98 to a Trp residue which is found in an endolysin from a bacteriophage of Listeria monocytogenes (PlyPSA). This mutation in CD27L resulted in an increased activity against selected serotypes of L. monocytogenes, demonstrating the potential to tune the species specificity of the catalytic domain of an endolysin. Structure-based modification of a Clostridium difficile-targeting endolysin affects activity and host range.,Mayer MJ, Garefalaki V, Spoerl R, Narbad A, Meijers R J Bacteriol. 2011 Oct;193(19):5477-86. Epub 2011 Jul 29. PMID:21803993 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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