Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.

Complement proteins can form membrane attack complex (MAC) pores that directly kill Gram-negative bacteria. MAC pores assemble by stepwise binding of C5b, C6, C7, C8 and finally C9, which can polymerize into a transmembrane ring of up to 18 C9 monomers. It is still unclear if the assembly of a polym...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Dennis J Doorduijn, Dani A C Heesterbeek, Maartje Ruyken, Carla J C de Haas, Daphne A C Stapels, Piet C Aerts, Suzan H M Rooijakkers, Bart W Bardoel
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/213580cfed6a4b06a4208b7d41522f83
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:213580cfed6a4b06a4208b7d41522f83
record_format	dspace
spelling	oai:doaj.org-article:213580cfed6a4b06a4208b7d41522f832021-12-02T19:59:53ZPolymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.1553-73661553-737410.1371/journal.ppat.1010051https://doaj.org/article/213580cfed6a4b06a4208b7d41522f832021-11-01T00:00:00Zhttps://doi.org/10.1371/journal.ppat.1010051https://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Complement proteins can form membrane attack complex (MAC) pores that directly kill Gram-negative bacteria. MAC pores assemble by stepwise binding of C5b, C6, C7, C8 and finally C9, which can polymerize into a transmembrane ring of up to 18 C9 monomers. It is still unclear if the assembly of a polymeric-C9 ring is necessary to sufficiently damage the bacterial cell envelope to kill bacteria. In this paper, polymerization of C9 was prevented without affecting binding of C9 to C5b-8, by locking the first transmembrane helix domain of C9. Using this system, we show that polymerization of C9 strongly enhanced damage to both the bacterial outer and inner membrane, resulting in more rapid killing of several Escherichia coli and Klebsiella strains in serum. By comparing binding of wildtype and 'locked' C9 by flow cytometry, we also show that polymerization of C9 is impaired when the amount of available C9 per C5b-8 is limited. This suggests that an excess of C9 is required to efficiently form polymeric-C9. Finally, we show that polymerization of C9 was impaired on complement-resistant E. coli strains that survive killing by MAC pores. This suggests that these bacteria can specifically block polymerization of C9. All tested complement-resistant E. coli expressed LPS O-antigen (O-Ag), compared to only one out of four complement-sensitive E. coli. By restoring O-Ag expression in an O-Ag negative strain, we show that the O-Ag impairs polymerization of C9 and results in complement-resistance. Altogether, these insights are important to understand how MAC pores kill bacteria and how bacterial pathogens can resist MAC-dependent killing.Dennis J DoorduijnDani A C HeesterbeekMaartje RuykenCarla J C de HaasDaphne A C StapelsPiet C AertsSuzan H M RooijakkersBart W BardoelPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 17, Iss 11, p e1010051 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 Dennis J Doorduijn Dani A C Heesterbeek Maartje Ruyken Carla J C de Haas Daphne A C Stapels Piet C Aerts Suzan H M Rooijakkers Bart W Bardoel Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
description	Complement proteins can form membrane attack complex (MAC) pores that directly kill Gram-negative bacteria. MAC pores assemble by stepwise binding of C5b, C6, C7, C8 and finally C9, which can polymerize into a transmembrane ring of up to 18 C9 monomers. It is still unclear if the assembly of a polymeric-C9 ring is necessary to sufficiently damage the bacterial cell envelope to kill bacteria. In this paper, polymerization of C9 was prevented without affecting binding of C9 to C5b-8, by locking the first transmembrane helix domain of C9. Using this system, we show that polymerization of C9 strongly enhanced damage to both the bacterial outer and inner membrane, resulting in more rapid killing of several Escherichia coli and Klebsiella strains in serum. By comparing binding of wildtype and 'locked' C9 by flow cytometry, we also show that polymerization of C9 is impaired when the amount of available C9 per C5b-8 is limited. This suggests that an excess of C9 is required to efficiently form polymeric-C9. Finally, we show that polymerization of C9 was impaired on complement-resistant E. coli strains that survive killing by MAC pores. This suggests that these bacteria can specifically block polymerization of C9. All tested complement-resistant E. coli expressed LPS O-antigen (O-Ag), compared to only one out of four complement-sensitive E. coli. By restoring O-Ag expression in an O-Ag negative strain, we show that the O-Ag impairs polymerization of C9 and results in complement-resistance. Altogether, these insights are important to understand how MAC pores kill bacteria and how bacterial pathogens can resist MAC-dependent killing.
format	article
author	Dennis J Doorduijn Dani A C Heesterbeek Maartje Ruyken Carla J C de Haas Daphne A C Stapels Piet C Aerts Suzan H M Rooijakkers Bart W Bardoel
author_facet	Dennis J Doorduijn Dani A C Heesterbeek Maartje Ruyken Carla J C de Haas Daphne A C Stapels Piet C Aerts Suzan H M Rooijakkers Bart W Bardoel
author_sort	Dennis J Doorduijn
title	Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
title_short	Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
title_full	Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
title_fullStr	Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
title_full_unstemmed	Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
title_sort	polymerization of c9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/213580cfed6a4b06a4208b7d41522f83
work_keys_str_mv	AT dennisjdoorduijn polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT daniacheesterbeek polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT maartjeruyken polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT carlajcdehaas polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT daphneacstapels polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT pietcaerts polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT suzanhmrooijakkers polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores AT bartwbardoel polymerizationofc9enhancesbacterialcellenvelopedamageandkillingbymembraneattackcomplexpores
_version_	1718375709292888064

Polymerization of C9 enhances bacterial cell envelope damage and killing by membrane attack complex pores.

Ejemplares similares