Journal Article

PUBDB-2015-01340

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

Zhang, G. (Corresponding Author) ; Mertens, H.>Extern*EMBL* ; Garefalaki, V. ; Jeffries, C.>Extern* ; Thompson, A. ; Lemke, E. A. ; Svergun, D.>Extern*EMBL* ; Mayer, M. J. ; Narbad, A. ; Meijers, R.>Extern*EMBL* ; Zhang, G.

2014
PLoS Lawrence, Kan.

This record in other databases:        

Please use a persistent id in citations: doi:10.1371/journal.ppat.1004228

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.

Note: OA

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Contributing Institute(s):

1. EMBL (EMBL)

Research Program(s):

1. DORIS Beamline K1.2 (POF2-54G13) (POF2-54G13)
2. PETRA Beamline P14 (POF2-54G14) (POF2-54G14)

Experiment(s):

1. DORIS Beamline K1.2 (DORIS III)
2. PETRA Beamline P14 (PETRA III)

Appears in the scientific report 2014

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Database coverage:
; ; ; ; BIOSIS Previews ; Current Contents - Life Sciences ; IF >= 5 ; JCR ; NCBI Molecular Biology Database ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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