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Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae

Carbapenemase and extended β-lactamase-producing Klebsiella pneumoniae isolates represent a major health threat, stimulating increasing interest in immunotherapeutic approaches for combating Klebsiella infections. Lipopolysaccharide O antigen polysaccharides offer viable targets for immunotherapeuti...

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Autores principales: Kelly, Steven D., Ovchinnikova, Olga G., Müller, Fabian, Steffen, Michael, Braun, Martin, Sweeney, Ryan P., Kowarik, Michael, Follador, Rainer, Lowary, Todd L., Serventi, Fabio, Whitfield, Chris
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10629545/
https://www.ncbi.nlm.nih.gov/pubmed/37428935
http://dx.doi.org/10.1073/pnas.2301302120
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author Kelly, Steven D.
Ovchinnikova, Olga G.
Müller, Fabian
Steffen, Michael
Braun, Martin
Sweeney, Ryan P.
Kowarik, Michael
Follador, Rainer
Lowary, Todd L.
Serventi, Fabio
Whitfield, Chris
author_facet Kelly, Steven D.
Ovchinnikova, Olga G.
Müller, Fabian
Steffen, Michael
Braun, Martin
Sweeney, Ryan P.
Kowarik, Michael
Follador, Rainer
Lowary, Todd L.
Serventi, Fabio
Whitfield, Chris
author_sort Kelly, Steven D.
collection PubMed
description Carbapenemase and extended β-lactamase-producing Klebsiella pneumoniae isolates represent a major health threat, stimulating increasing interest in immunotherapeutic approaches for combating Klebsiella infections. Lipopolysaccharide O antigen polysaccharides offer viable targets for immunotherapeutic development, and several studies have described protection with O-specific antibodies in animal models of infection. O1 antigen is produced by almost half of clinical Klebsiella isolates. The O1 polysaccharide backbone structure is known, but monoclonal antibodies raised against the O1 antigen showed varying reactivity against different isolates that could not be explained by the known structure. Reinvestigation of the structure by NMR spectroscopy revealed the presence of the reported polysaccharide backbone (glycoform O1a), as well as a previously unknown O1b glycoform composed of the O1a backbone modified with a terminal pyruvate group. The activity of the responsible pyruvyltransferase (WbbZ) was confirmed by western immunoblotting and in vitro chemoenzymatic synthesis of the O1b terminus. Bioinformatic data indicate that almost all O1 isolates possess genes required to produce both glycoforms. We describe the presence of O1ab-biosynthesis genes in other bacterial species and report a functional O1 locus on a bacteriophage genome. Homologs of wbbZ are widespread in genetic loci for the assembly of unrelated glycostructures in bacteria and yeast. In K. pneumoniae, simultaneous production of both O1 glycoforms is enabled by the lack of specificity of the ABC transporter that exports the nascent glycan, and the data reported here provide mechanistic understanding of the capacity for evolution of antigenic diversity within an important class of biomolecules produced by many bacteria.
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spelling pubmed-106295452023-11-08 Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae Kelly, Steven D. Ovchinnikova, Olga G. Müller, Fabian Steffen, Michael Braun, Martin Sweeney, Ryan P. Kowarik, Michael Follador, Rainer Lowary, Todd L. Serventi, Fabio Whitfield, Chris Proc Natl Acad Sci U S A Biological Sciences Carbapenemase and extended β-lactamase-producing Klebsiella pneumoniae isolates represent a major health threat, stimulating increasing interest in immunotherapeutic approaches for combating Klebsiella infections. Lipopolysaccharide O antigen polysaccharides offer viable targets for immunotherapeutic development, and several studies have described protection with O-specific antibodies in animal models of infection. O1 antigen is produced by almost half of clinical Klebsiella isolates. The O1 polysaccharide backbone structure is known, but monoclonal antibodies raised against the O1 antigen showed varying reactivity against different isolates that could not be explained by the known structure. Reinvestigation of the structure by NMR spectroscopy revealed the presence of the reported polysaccharide backbone (glycoform O1a), as well as a previously unknown O1b glycoform composed of the O1a backbone modified with a terminal pyruvate group. The activity of the responsible pyruvyltransferase (WbbZ) was confirmed by western immunoblotting and in vitro chemoenzymatic synthesis of the O1b terminus. Bioinformatic data indicate that almost all O1 isolates possess genes required to produce both glycoforms. We describe the presence of O1ab-biosynthesis genes in other bacterial species and report a functional O1 locus on a bacteriophage genome. Homologs of wbbZ are widespread in genetic loci for the assembly of unrelated glycostructures in bacteria and yeast. In K. pneumoniae, simultaneous production of both O1 glycoforms is enabled by the lack of specificity of the ABC transporter that exports the nascent glycan, and the data reported here provide mechanistic understanding of the capacity for evolution of antigenic diversity within an important class of biomolecules produced by many bacteria. National Academy of Sciences 2023-07-10 2023-07-18 /pmc/articles/PMC10629545/ /pubmed/37428935 http://dx.doi.org/10.1073/pnas.2301302120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Biological Sciences
Kelly, Steven D.
Ovchinnikova, Olga G.
Müller, Fabian
Steffen, Michael
Braun, Martin
Sweeney, Ryan P.
Kowarik, Michael
Follador, Rainer
Lowary, Todd L.
Serventi, Fabio
Whitfield, Chris
Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title_full Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title_fullStr Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title_full_unstemmed Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title_short Identification of a second glycoform of the clinically prevalent O1 antigen from Klebsiella pneumoniae
title_sort identification of a second glycoform of the clinically prevalent o1 antigen from klebsiella pneumoniae
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10629545/
https://www.ncbi.nlm.nih.gov/pubmed/37428935
http://dx.doi.org/10.1073/pnas.2301302120
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