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@ARTICLE{Zhang:207430,
author = {Zhang, Gongyi and Mertens, Haydyn and Garefalaki, Vasiliki
and Jeffries, Cy and Thompson, Andrew and Lemke, Edward A.
and Svergun, Dmitri and Mayer, Melinda J. and Narbad, Arjan
and Meijers, Rob and Zhang, Gongyi},
title = {{T}he {CD}27{L} and {CTP}1{L} {E}ndolysins {T}argeting
{C}lostridia {C}ontain a {B}uilt-in {T}rigger and {R}elease
{F}actor},
journal = {PLoS pathogens},
volume = {10},
number = {7},
issn = {1553-7374},
address = {Lawrence, Kan.},
publisher = {PLoS},
reportid = {PUBDB-2015-01340},
pages = {e1004228 -},
year = {2014},
note = {OA},
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, 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.},
cin = {EMBL},
ddc = {610},
cid = {I:(DE-H253)EMBL-20120731},
pnm = {DORIS Beamline K1.2 (POF2-54G13) / PETRA Beamline P14
(POF2-54G14)},
pid = {G:(DE-H253)POF2-K1.2-20130405 /
G:(DE-H253)POF2-P14-20130405},
experiment = {EXP:(DE-H253)D-K1.2-20150101 / EXP:(DE-H253)P-P14-20150101},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000340551000023},
pubmed = {pmid:25058163},
doi = {10.1371/journal.ppat.1004228},
url = {https://bib-pubdb1.desy.de/record/207430},
}