Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	The CD27L and CTP1L endolysins targeting Clostridia contain a built-in trigger and release factor.
Journal, issue, pages	PLoS Pathog, Vol. 10, Issue 7, Page e1004228, Year 2014
Publish date	Jul 24, 2014
Authors	Matthew Dunne / Haydyn D T Mertens / Vasiliki Garefalaki / Cy M Jeffries / Andrew Thompson / Edward A Lemke / Dmitri I Svergun / Melinda J Mayer / Arjan Narbad / Rob Meijers /
PubMed Abstract	The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, ...The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, and represents a promising alternative as a bactericide. To better understand the lysis mechanism, we have determined the crystal structure of an autoproteolytic fragment of the CD27L endolysin. The structure covers the C-terminal domain of the endolysin, and represents a novel fold that is identified in a number of lysins that target Clostridia bacteria. The structure indicates endolysin cleavage occurs at the stem of the linker connecting the catalytic domain with the C-terminal domain. We also solved the crystal structure of the C-terminal domain of a slow cleaving mutant of the CTP1L endolysin that targets C. tyrobutyricum. Two distinct dimerization modes are observed in the crystal structures for both endolysins, despite a sequence identity of only 22% between the domains. The dimers are validated to be present for the full length protein in solution by right angle light scattering, small angle X-ray scattering and cross-linking experiments using the cross-linking amino acid p-benzoyl-L-phenylalanine (pBpa). Mutagenesis on residues contributing to the dimer interfaces indicates that there is a link between the dimerization modes and the autocleavage mechanism. We show that for the CTP1L endolysin, there is a reduction in lysis efficiency that is proportional to the cleavage efficiency. We propose a model for endolysin triggering, where the extended dimer presents the inactive state, and a switch to the side-by-side dimer triggers the cleavage of the C-terminal domain. This leads to the release of the catalytic portion of the endolysin, enabling the efficient digestion of the bacterial cell wall.
External links	PLoS Pathog / PubMed:25058163 / PubMed Central
Methods	SAS (X-ray synchrotron) / X-ray diffraction
Resolution	2.11 - 2.24 Å
Structure data	SASDAL5: CD27L (Clostridium difficile bacteriophage 27 endolysin) Method: SAXS/SANS SASDAM5: CD27L (C238R) (Clostridium difficile bacteriophage 27 endolysin C238R mutant, CD27L(C238R)) Method: SAXS/SANS PDB-4cu2: C-terminal domain of CTP1L endolysin mutant V195P that reduces autoproteolysis Method: X-RAY DIFFRACTION / Resolution: 2.11 Å PDB-4cu5: C-terminal domain of endolysin from phage CD27L is a trigger and release factor Method: X-RAY DIFFRACTION / Resolution: 2.24 Å
Chemicals	ChemComp-HOH: WATER
Source	Clostridioides difficile (bacteria) clostridium phage phictp1 (virus) clostridium phage phicd27 (virus)
Keywords	HYDROLASE / BACTERIAL LYSIS / AUTOPROTEOLYSIS / BACTERIOPHAGE