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The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing

The rising antimicrobial resistance is particularly alarming for Acinetobacter baumannii, calling for the discovery and evaluation of alternatives to treat A. baumannii infections. Some bacteriophages produce a structural protein that depolymerizes capsular exopolysaccharide. Such purified depolymer...

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Autores principales: Abdelkader, Karim, Gutiérrez, Diana, Latka, Agnieszka, Boeckaerts, Dimitri, Drulis-Kawa, Zuzanna, Criel, Bjorn, Gerstmans, Hans, Safaan, Amal, Khairalla, Ahmed S., Gaber, Yasser, Dishisha, Tarek, Briers, Yves
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9137491/
https://www.ncbi.nlm.nih.gov/pubmed/35625321
http://dx.doi.org/10.3390/antibiotics11050677
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author Abdelkader, Karim
Gutiérrez, Diana
Latka, Agnieszka
Boeckaerts, Dimitri
Drulis-Kawa, Zuzanna
Criel, Bjorn
Gerstmans, Hans
Safaan, Amal
Khairalla, Ahmed S.
Gaber, Yasser
Dishisha, Tarek
Briers, Yves
author_facet Abdelkader, Karim
Gutiérrez, Diana
Latka, Agnieszka
Boeckaerts, Dimitri
Drulis-Kawa, Zuzanna
Criel, Bjorn
Gerstmans, Hans
Safaan, Amal
Khairalla, Ahmed S.
Gaber, Yasser
Dishisha, Tarek
Briers, Yves
author_sort Abdelkader, Karim
collection PubMed
description The rising antimicrobial resistance is particularly alarming for Acinetobacter baumannii, calling for the discovery and evaluation of alternatives to treat A. baumannii infections. Some bacteriophages produce a structural protein that depolymerizes capsular exopolysaccharide. Such purified depolymerases are considered as novel antivirulence compounds. We identified and characterized a depolymerase (DpoMK34) from Acinetobacter phage vB_AbaP_PMK34 active against the clinical isolate A. baumannii MK34. In silico analysis reveals a modular protein displaying a conserved N-terminal domain for anchoring to the phage tail, and variable central and C-terminal domains for enzymatic activity and specificity. AlphaFold-Multimer predicts a trimeric protein adopting an elongated structure due to a long α-helix, an enzymatic β-helix domain and a hypervariable 4 amino acid hotspot in the most ultimate loop of the C-terminal domain. In contrast to the tail fiber of phage T3, this hypervariable hotspot appears unrelated with the primary receptor. The functional characterization of DpoMK34 revealed a mesophilic enzyme active up to 50 °C across a wide pH range (4 to 11) and specific for the capsule of A. baumannii MK34. Enzymatic degradation of the A. baumannii MK34 capsule causes a significant drop in phage adsorption from 95% to 9% after 5 min. Although lacking intrinsic antibacterial activity, DpoMK34 renders A. baumannii MK34 fully susceptible to serum killing in a serum concentration dependent manner. Unlike phage PMK34, DpoMK34 does not easily select for resistant mutants either against PMK34 or itself. In sum, DpoMK34 is a potential antivirulence compound that can be included in a depolymerase cocktail to control difficult to treat A. baumannii infections.
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spelling pubmed-91374912022-05-28 The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing Abdelkader, Karim Gutiérrez, Diana Latka, Agnieszka Boeckaerts, Dimitri Drulis-Kawa, Zuzanna Criel, Bjorn Gerstmans, Hans Safaan, Amal Khairalla, Ahmed S. Gaber, Yasser Dishisha, Tarek Briers, Yves Antibiotics (Basel) Article The rising antimicrobial resistance is particularly alarming for Acinetobacter baumannii, calling for the discovery and evaluation of alternatives to treat A. baumannii infections. Some bacteriophages produce a structural protein that depolymerizes capsular exopolysaccharide. Such purified depolymerases are considered as novel antivirulence compounds. We identified and characterized a depolymerase (DpoMK34) from Acinetobacter phage vB_AbaP_PMK34 active against the clinical isolate A. baumannii MK34. In silico analysis reveals a modular protein displaying a conserved N-terminal domain for anchoring to the phage tail, and variable central and C-terminal domains for enzymatic activity and specificity. AlphaFold-Multimer predicts a trimeric protein adopting an elongated structure due to a long α-helix, an enzymatic β-helix domain and a hypervariable 4 amino acid hotspot in the most ultimate loop of the C-terminal domain. In contrast to the tail fiber of phage T3, this hypervariable hotspot appears unrelated with the primary receptor. The functional characterization of DpoMK34 revealed a mesophilic enzyme active up to 50 °C across a wide pH range (4 to 11) and specific for the capsule of A. baumannii MK34. Enzymatic degradation of the A. baumannii MK34 capsule causes a significant drop in phage adsorption from 95% to 9% after 5 min. Although lacking intrinsic antibacterial activity, DpoMK34 renders A. baumannii MK34 fully susceptible to serum killing in a serum concentration dependent manner. Unlike phage PMK34, DpoMK34 does not easily select for resistant mutants either against PMK34 or itself. In sum, DpoMK34 is a potential antivirulence compound that can be included in a depolymerase cocktail to control difficult to treat A. baumannii infections. MDPI 2022-05-17 /pmc/articles/PMC9137491/ /pubmed/35625321 http://dx.doi.org/10.3390/antibiotics11050677 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Abdelkader, Karim
Gutiérrez, Diana
Latka, Agnieszka
Boeckaerts, Dimitri
Drulis-Kawa, Zuzanna
Criel, Bjorn
Gerstmans, Hans
Safaan, Amal
Khairalla, Ahmed S.
Gaber, Yasser
Dishisha, Tarek
Briers, Yves
The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title_full The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title_fullStr The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title_full_unstemmed The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title_short The Specific Capsule Depolymerase of Phage PMK34 Sensitizes Acinetobacter baumannii to Serum Killing
title_sort specific capsule depolymerase of phage pmk34 sensitizes acinetobacter baumannii to serum killing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9137491/
https://www.ncbi.nlm.nih.gov/pubmed/35625321
http://dx.doi.org/10.3390/antibiotics11050677
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