A stable phage lysin (Cpl-1) dimer with increased antipneumococcal activity and decreased plasma clearance

Resch, Grégory; Moreillon, Philippe; Fischetti, Vincent A.

doi:10.1016/j.ijantimicag.2011.08.009

Cited by 46 publications

(50 citation statements)

References 32 publications

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“…The pneumococcal autolysin LytA does dimerize into a conformation resembling the side-by-side dimer presented here, and it was suggested that the dimer conformation may contribute to its activity [26]. The CPL-1 phage endolysin that targets Streptococcus pneumoniae was engineered to stabilize the (what we call) side-by-side dimerization mode, and this led to a ten fold increase in its activity [27].…”

Section: Discussionmentioning

confidence: 69%

The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

et al. 2014

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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.

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Section: Discussionmentioning

confidence: 69%

The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

et al. 2014

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“…This will mean that EC300 assembles one M23 endopeptidase CD with four copies of the CWB170, which provides to the lytic enzyme high affinity to the cell wall (Proença et al 2014). In addition, due to its multimeric nature, EC300 is a protein of almost 70 kDa; this will certainly be an advantage for the study of its effectiveness in animal infection models since, as observed for the dimeric form of the pneumococcal endolysin Cpl-1 (Resch et al 2011), it should reduce renal clearance (proteins smaller than 60-65 kDa tend to be rapidly eliminated by glomerular filtration in humans; Maack et al 1979).…”

Section: Discussionmentioning

confidence: 99%

EC300: a phage-based, bacteriolysin-like protein with enhanced antibacterial activity against Enterococcus faecalis

Proença

Leandro

García

et al. 2015

Appl Microbiol Biotechnol

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Bacteriophage lytic enzymes, either endolysins or virion-associated lysins, have been receiving considerable attention as potential antibacterial agents, particularly for the combat of antibiotic-resistant Gram-positive pathogens. A conclusion that easily emerges from the careful analysis of a great number of reports on the field is that the activity of phage lytic enzymes is rarely studied in conditions that support robust growth of the target bacteria. Here, we report the construction and study of a chimerical lysin, EC300, which was designed to target and kill Enterococcus faecalis in conditions supporting vigorous bacterial growth. EC300 resulted from the fusion of a predicted M23 endopeptidase domain of a virion-associated lysin to the putative cell wall binding domain of a previously characterized amidase endolysin, both produced by the E. faecalis phage F170/08. This bacteriolysin-like protein exhibited a clear enhanced lytic activity over the parental endolysin when both were assayed in a rich bacterial growth medium. We demonstrate the killing efficacy of EC300 against growing cells of a panel of typed E. faecalis clinical strains with high level of antibiotic resistance. The possible reasons for the marked difference between the lytic performance of EC300 and that of the amidase are discussed.

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“…In the last decade, several staphylococci targeted lysins have been developed and examined in experimental animal models to validate their efficacy as therapeutic agents14. However, natural staphylococcal lysins frequently suffer from solubility/stability and activity problems1516. Moreover, the activities of some staphylococcal lysins are evidenced to be affected by working conditions, such as milk17, and ions concentrations18.…”

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confidence: 99%

A novel chimeric lysin with robust antibacterial activity against planktonic and biofilm methicillin-resistant Staphylococcus aureus

Yang

Zhang

Wang

et al. 2017

Sci Rep

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Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most threatening pathogens due to its multi-drug resistance (MDR) and strong biofilm-forming capacity. Here, we described the screening of a novel chimeolysin (ClyF) that was active against planktonic and biofilm MRSA. Biochemical tests showed that ClyF was active against all S. aureus clinical isolates tested under planktonic and biofilm conditions. Structure analysis revealed that ClyF has an enhanced thermostability and pH tolerance than its parental lysin Pc by forming a hydrophobic cleft in the catalytic domain and an Ig-like structure in the cell-wall binding domain. A single intraperitoneally or topically administration of ClyF showed good MRSA removing efficacy in mouse models of bacteremia and burn wound infection, respectively. Our data collectively demonstrated that ClyF has good bactericidal activity against planktonic and biofilm MRSA both in vitro and in vivo, and therefore represents a useful antibacterial to combat MDR S. aureus.

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A stable phage lysin (Cpl-1) dimer with increased antipneumococcal activity and decreased plasma clearance

Cited by 46 publications

References 32 publications

The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

EC300: a phage-based, bacteriolysin-like protein with enhanced antibacterial activity against Enterococcus faecalis

A novel chimeric lysin with robust antibacterial activity against planktonic and biofilm methicillin-resistant Staphylococcus aureus

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