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PEGylating a bacteriophage endolysin inhibits its bactericidal activity
Bacteriophage endolysins (lysins) bind to a cell wall substrate and cleave peptidoglycan, resulting in hypotonic lysis of the phage-infected bacteria. When purified lysins are added externally to Gram-positive bacteria they mediate rapid death by the same mechanism. For this reason, novel therapeuti...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3222324/ https://www.ncbi.nlm.nih.gov/pubmed/21982426 http://dx.doi.org/10.1186/2191-0855-1-29 |
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author | Resch, Gregory Moreillon, Philippe Fischetti, Vincent A |
author_facet | Resch, Gregory Moreillon, Philippe Fischetti, Vincent A |
author_sort | Resch, Gregory |
collection | PubMed |
description | Bacteriophage endolysins (lysins) bind to a cell wall substrate and cleave peptidoglycan, resulting in hypotonic lysis of the phage-infected bacteria. When purified lysins are added externally to Gram-positive bacteria they mediate rapid death by the same mechanism. For this reason, novel therapeutic strategies have been developed using such enzybiotics. However, like other proteins introduced into mammalian organisms, they are quickly cleared from systemic circulation. PEGylation has been used successfully to increase the in vivo half-life of many biological molecules and was therefore applied to Cpl-1, a lysin specific for S. pneumoniae. Cysteine-specific PEGylation with either PEG 10K or 40K was achieved on Cpl-1 mutants, each containing an additional cysteine residue at different locations To the best of our knowledge, this is the first report of the PEGylation of bacteriophage lysin. Compared to the native enzyme, none of the PEGylated conjugates retained significant in vitro anti-pneumococcal lytic activity that would have justified further in vivo studies. Since the anti-microbial activity of the mutant enzymes used in this study was not affected by the introduction of the cysteine residue, our results implied that the presence of the PEG molecule was responsible for the inhibition. As most endolysins exhibit a similar modular structure, we believe that our work emphasizes the inability to improve the in vivo half-life of this class of enzybiotics using a cysteine-specific PEGylation strategy. |
format | Online Article Text |
id | pubmed-3222324 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Springer |
record_format | MEDLINE/PubMed |
spelling | pubmed-32223242011-12-16 PEGylating a bacteriophage endolysin inhibits its bactericidal activity Resch, Gregory Moreillon, Philippe Fischetti, Vincent A AMB Express Original Bacteriophage endolysins (lysins) bind to a cell wall substrate and cleave peptidoglycan, resulting in hypotonic lysis of the phage-infected bacteria. When purified lysins are added externally to Gram-positive bacteria they mediate rapid death by the same mechanism. For this reason, novel therapeutic strategies have been developed using such enzybiotics. However, like other proteins introduced into mammalian organisms, they are quickly cleared from systemic circulation. PEGylation has been used successfully to increase the in vivo half-life of many biological molecules and was therefore applied to Cpl-1, a lysin specific for S. pneumoniae. Cysteine-specific PEGylation with either PEG 10K or 40K was achieved on Cpl-1 mutants, each containing an additional cysteine residue at different locations To the best of our knowledge, this is the first report of the PEGylation of bacteriophage lysin. Compared to the native enzyme, none of the PEGylated conjugates retained significant in vitro anti-pneumococcal lytic activity that would have justified further in vivo studies. Since the anti-microbial activity of the mutant enzymes used in this study was not affected by the introduction of the cysteine residue, our results implied that the presence of the PEG molecule was responsible for the inhibition. As most endolysins exhibit a similar modular structure, we believe that our work emphasizes the inability to improve the in vivo half-life of this class of enzybiotics using a cysteine-specific PEGylation strategy. Springer 2011-10-07 /pmc/articles/PMC3222324/ /pubmed/21982426 http://dx.doi.org/10.1186/2191-0855-1-29 Text en Copyright ©2011 Resch et al; licensee Springer. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Resch, Gregory Moreillon, Philippe Fischetti, Vincent A PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title | PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title_full | PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title_fullStr | PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title_full_unstemmed | PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title_short | PEGylating a bacteriophage endolysin inhibits its bactericidal activity |
title_sort | pegylating a bacteriophage endolysin inhibits its bactericidal activity |
topic | Original |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3222324/ https://www.ncbi.nlm.nih.gov/pubmed/21982426 http://dx.doi.org/10.1186/2191-0855-1-29 |
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