The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins

ABSTRACT Glycans decorate proteins and affect their biological function, including protection against proteolytic degradation. However, pathogenic, and commensal bacteria have evolved specific glycoproteases that overcome the steric impediment posed by carbohydrates, cleaving glycoproteins precisely...

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Autores principales:	M. Florencia Haurat, Nichollas E. Scott, Gisela Di Venanzio, Juvenal Lopez, Benjamin Pluvinage, Alisdair B. Boraston, Michael J. Ferracane, Mario F. Feldman
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Acinetobacter CpaA T2SS glycobiology glycoprotease Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/77f8b47752434d51900a4f72a46fcc2f
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spelling	oai:doaj.org-article:77f8b47752434d51900a4f72a46fcc2f2021-11-15T16:19:09ZThe Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins10.1128/mBio.02033-202150-7511https://doaj.org/article/77f8b47752434d51900a4f72a46fcc2f2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02033-20https://doaj.org/toc/2150-7511ABSTRACT Glycans decorate proteins and affect their biological function, including protection against proteolytic degradation. However, pathogenic, and commensal bacteria have evolved specific glycoproteases that overcome the steric impediment posed by carbohydrates, cleaving glycoproteins precisely at their glycosylation site(s). Medically relevant Acinetobacter strains employ their type II secretion system (T2SS) to secrete the glycoprotease CpaA, which contributes to virulence. Previously, CpaA was shown to cleave two O-linked glycoproteins, factors V and XII, leading to reduced blood coagulation. In this work, we show that CpaA cleaves a broader range of O-linked human glycoproteins, including several glycoproteins involved in complement activation, such as CD55 and CD46. However, only CD55 was removed from the cell surface, while CD46 remained unaltered during the Acinetobacter nosocomialis infection assay. We show that CpaA has a unique consensus target sequence that consists of a glycosylated serine or threonine residue after a proline residue (P-S/T), and its activity is not affected by sialic acids. Molecular modeling and mutagenesis analysis of CpaA suggest that the indole ring of Trp493 and the ring of the Pro residue in the substrate form a key interaction that contributes to CpaA sequence selectivity. Similar bacterial glycoproteases have recently gained attention as tools for proteomic analysis of human glycoproteins, and CpaA appears to be a robust and attractive new component of the glycoproteomics toolbox. Combined, our work provides insight into the function and possible application of CpaA, a member of a widespread class of broad-spectrum bacterial glycoproteases involved in host-pathogen interactions. IMPORTANCE CpaA is a glycoprotease expressed by members of the Acinetobacter baumannii-calcoaceticus complex, and it is the first bona fide secreted virulence factor identified in these species. Here, we show that CpaA cleaves multiple targets precisely at O-glycosylation sites preceded by a Pro residue. This feature, together with the observation that sialic acid does not impact CpaA activity, makes this enzyme an attractive tool for the analysis of O-linked human protein for biotechnical and diagnostic purposes. Previous work identified proteins involved in blood coagulation as targets of CpaA. Our work broadens the set of targets of CpaA, pointing toward additional roles in bacterium-host interactions. We propose that CpaA belongs to an expanding class of functionally defined glycoproteases that targets multiple O-linked host glycoproteins.M. Florencia HauratNichollas E. ScottGisela Di VenanzioJuvenal LopezBenjamin PluvinageAlisdair B. BorastonMichael J. FerracaneMario F. FeldmanAmerican Society for MicrobiologyarticleAcinetobacterCpaAT2SSglycobiologyglycoproteaseMicrobiologyQR1-502ENmBio, Vol 11, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Acinetobacter CpaA T2SS glycobiology glycoprotease Microbiology QR1-502
spellingShingle	Acinetobacter CpaA T2SS glycobiology glycoprotease Microbiology QR1-502 M. Florencia Haurat Nichollas E. Scott Gisela Di Venanzio Juvenal Lopez Benjamin Pluvinage Alisdair B. Boraston Michael J. Ferracane Mario F. Feldman The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
description	ABSTRACT Glycans decorate proteins and affect their biological function, including protection against proteolytic degradation. However, pathogenic, and commensal bacteria have evolved specific glycoproteases that overcome the steric impediment posed by carbohydrates, cleaving glycoproteins precisely at their glycosylation site(s). Medically relevant Acinetobacter strains employ their type II secretion system (T2SS) to secrete the glycoprotease CpaA, which contributes to virulence. Previously, CpaA was shown to cleave two O-linked glycoproteins, factors V and XII, leading to reduced blood coagulation. In this work, we show that CpaA cleaves a broader range of O-linked human glycoproteins, including several glycoproteins involved in complement activation, such as CD55 and CD46. However, only CD55 was removed from the cell surface, while CD46 remained unaltered during the Acinetobacter nosocomialis infection assay. We show that CpaA has a unique consensus target sequence that consists of a glycosylated serine or threonine residue after a proline residue (P-S/T), and its activity is not affected by sialic acids. Molecular modeling and mutagenesis analysis of CpaA suggest that the indole ring of Trp493 and the ring of the Pro residue in the substrate form a key interaction that contributes to CpaA sequence selectivity. Similar bacterial glycoproteases have recently gained attention as tools for proteomic analysis of human glycoproteins, and CpaA appears to be a robust and attractive new component of the glycoproteomics toolbox. Combined, our work provides insight into the function and possible application of CpaA, a member of a widespread class of broad-spectrum bacterial glycoproteases involved in host-pathogen interactions. IMPORTANCE CpaA is a glycoprotease expressed by members of the Acinetobacter baumannii-calcoaceticus complex, and it is the first bona fide secreted virulence factor identified in these species. Here, we show that CpaA cleaves multiple targets precisely at O-glycosylation sites preceded by a Pro residue. This feature, together with the observation that sialic acid does not impact CpaA activity, makes this enzyme an attractive tool for the analysis of O-linked human protein for biotechnical and diagnostic purposes. Previous work identified proteins involved in blood coagulation as targets of CpaA. Our work broadens the set of targets of CpaA, pointing toward additional roles in bacterium-host interactions. We propose that CpaA belongs to an expanding class of functionally defined glycoproteases that targets multiple O-linked host glycoproteins.
format	article
author	M. Florencia Haurat Nichollas E. Scott Gisela Di Venanzio Juvenal Lopez Benjamin Pluvinage Alisdair B. Boraston Michael J. Ferracane Mario F. Feldman
author_facet	M. Florencia Haurat Nichollas E. Scott Gisela Di Venanzio Juvenal Lopez Benjamin Pluvinage Alisdair B. Boraston Michael J. Ferracane Mario F. Feldman
author_sort	M. Florencia Haurat
title	The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
title_short	The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
title_full	The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
title_fullStr	The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
title_full_unstemmed	The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins
title_sort	glycoprotease cpaa secreted by medically relevant <named-content content-type="genus-species">acinetobacter</named-content> species targets multiple <italic toggle="yes">o</italic>-linked host glycoproteins
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/77f8b47752434d51900a4f72a46fcc2f
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The Glycoprotease CpaA Secreted by Medically Relevant <named-content content-type="genus-species">Acinetobacter</named-content> Species Targets Multiple <italic toggle="yes">O</italic>-Linked Host Glycoproteins

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