Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins

ABSTRACT The posttranslational Ca2+-dependent “clip-and-link” activity of large repeat-in-toxin (RTX) proteins starts by Ca2+-dependent structural rearrangement of a highly conserved self-processing module (SPM). Subsequently, an internal aspartate-proline (Asp-Pro) peptide bond at the N-terminal en...

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Autores principales:	Vojtech Kuban, Pavel Macek, Jozef Hritz, Katerina Nechvatalova, Katerina Nedbalcova, Martin Faldyna, Peter Sebo, Lukas Zidek, Ladislav Bumba
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	RTX toxins cell adhesion clip-and-link host-pathogen interactions nuclear magnetic resonance Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/fdc4a6e79d8040c09e2fbb26185124d6
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spelling	oai:doaj.org-article:fdc4a6e79d8040c09e2fbb26185124d62021-11-15T15:57:02ZStructural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins10.1128/mBio.00226-202150-7511https://doaj.org/article/fdc4a6e79d8040c09e2fbb26185124d62020-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00226-20https://doaj.org/toc/2150-7511ABSTRACT The posttranslational Ca2+-dependent “clip-and-link” activity of large repeat-in-toxin (RTX) proteins starts by Ca2+-dependent structural rearrangement of a highly conserved self-processing module (SPM). Subsequently, an internal aspartate-proline (Asp-Pro) peptide bond at the N-terminal end of SPM breaks, and the liberated C-terminal aspartyl residue can react with a free ε-amino group of an adjacent lysine residue to form a new isopeptide bond. Here, we report a solution structure of the calcium-loaded SPM (Ca-SPM) derived from the FrpC protein of Neisseria meningitidis. The Ca-SPM structure defines a unique protein architecture and provides structural insight into the autocatalytic cleavage of the Asp-Pro peptide bond through a “twisted-amide” activation. Furthermore, in-frame deletion of the SPM domain from the ApxIVA protein of Actinobacillus pleuropneumoniae attenuated the virulence of this porcine pathogen in a pig respiratory challenge model. We hypothesize that the Ca2+-dependent clip-and-link activity represents an unconventional strategy for Gram-negative pathogens to adhere to the host target cell surface. IMPORTANCE The Ca2+-dependent clip-and-link activity of large repeat-in-toxin (RTX) proteins is an exceptional posttranslational process in which an internal domain called a self-processing module (SPM) mediates Ca2+-dependent processing of a highly specific aspartate-proline (Asp-Pro) peptide bond and covalent linkage of the released aspartyl to an adjacent lysine residue through an isopeptide bond. Here, we report the solution structures of the Ca2+-loaded SPM (Ca-SPM) defining the mechanism of the autocatalytic cleavage of the Asp414-Pro415 peptide bond of the Neisseria meningitidis FrpC exoprotein. Moreover, deletion of the SPM domain in the ApxIVA protein, the FrpC homolog of Actinobacillus pleuropneumoniae, resulted in attenuation of virulence of the bacterium in a pig infection model, indicating that the Ca2+-dependent clip-and-link activity plays a role in the virulence of Gram-negative pathogens.Vojtech KubanPavel MacekJozef HritzKaterina NechvatalovaKaterina NedbalcovaMartin FaldynaPeter SeboLukas ZidekLadislav BumbaAmerican Society for MicrobiologyarticleRTX toxinscell adhesionclip-and-linkhost-pathogen interactionsnuclear magnetic resonanceMicrobiologyQR1-502ENmBio, Vol 11, Iss 2 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	RTX toxins cell adhesion clip-and-link host-pathogen interactions nuclear magnetic resonance Microbiology QR1-502
spellingShingle	RTX toxins cell adhesion clip-and-link host-pathogen interactions nuclear magnetic resonance Microbiology QR1-502 Vojtech Kuban Pavel Macek Jozef Hritz Katerina Nechvatalova Katerina Nedbalcova Martin Faldyna Peter Sebo Lukas Zidek Ladislav Bumba Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
description	ABSTRACT The posttranslational Ca2+-dependent “clip-and-link” activity of large repeat-in-toxin (RTX) proteins starts by Ca2+-dependent structural rearrangement of a highly conserved self-processing module (SPM). Subsequently, an internal aspartate-proline (Asp-Pro) peptide bond at the N-terminal end of SPM breaks, and the liberated C-terminal aspartyl residue can react with a free ε-amino group of an adjacent lysine residue to form a new isopeptide bond. Here, we report a solution structure of the calcium-loaded SPM (Ca-SPM) derived from the FrpC protein of Neisseria meningitidis. The Ca-SPM structure defines a unique protein architecture and provides structural insight into the autocatalytic cleavage of the Asp-Pro peptide bond through a “twisted-amide” activation. Furthermore, in-frame deletion of the SPM domain from the ApxIVA protein of Actinobacillus pleuropneumoniae attenuated the virulence of this porcine pathogen in a pig respiratory challenge model. We hypothesize that the Ca2+-dependent clip-and-link activity represents an unconventional strategy for Gram-negative pathogens to adhere to the host target cell surface. IMPORTANCE The Ca2+-dependent clip-and-link activity of large repeat-in-toxin (RTX) proteins is an exceptional posttranslational process in which an internal domain called a self-processing module (SPM) mediates Ca2+-dependent processing of a highly specific aspartate-proline (Asp-Pro) peptide bond and covalent linkage of the released aspartyl to an adjacent lysine residue through an isopeptide bond. Here, we report the solution structures of the Ca2+-loaded SPM (Ca-SPM) defining the mechanism of the autocatalytic cleavage of the Asp414-Pro415 peptide bond of the Neisseria meningitidis FrpC exoprotein. Moreover, deletion of the SPM domain in the ApxIVA protein, the FrpC homolog of Actinobacillus pleuropneumoniae, resulted in attenuation of virulence of the bacterium in a pig infection model, indicating that the Ca2+-dependent clip-and-link activity plays a role in the virulence of Gram-negative pathogens.
format	article
author	Vojtech Kuban Pavel Macek Jozef Hritz Katerina Nechvatalova Katerina Nedbalcova Martin Faldyna Peter Sebo Lukas Zidek Ladislav Bumba
author_facet	Vojtech Kuban Pavel Macek Jozef Hritz Katerina Nechvatalova Katerina Nedbalcova Martin Faldyna Peter Sebo Lukas Zidek Ladislav Bumba
author_sort	Vojtech Kuban
title	Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
title_short	Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
title_full	Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
title_fullStr	Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
title_full_unstemmed	Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
title_sort	structural basis of ca<sup>2+</sup>-dependent self-processing activity of repeat-in-toxin proteins
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/fdc4a6e79d8040c09e2fbb26185124d6
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Structural Basis of Ca<sup>2+</sup>-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins

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