Structural Basis of Ca2+-Dependent Self-Processing Activity of
Repeat-in-Toxin Proteins

J 2020

Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins

KUBÁŇ, Vojtěch, P. MACEK, Jozef HRITZ, K. NECHVATALOVA, K. NEDBALCOVA et. al.

Basic information

Original name

Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins

Authors

KUBÁŇ, Vojtěch (203 Czech Republic, belonging to the institution), P. MACEK, Jozef HRITZ (703 Slovakia, belonging to the institution), K. NECHVATALOVA, K. NEDBALCOVA, M. FALDYNA, Peter ŠEBO (203 Czech Republic, belonging to the institution), Lukáš ŽÍDEK (203 Czech Republic, guarantor, belonging to the institution) and L. BUMBA

Edition

MBIO, Washington, D.C. American Society for Microbiology, 2020, 2150-7511

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10606 Microbiology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 7.867

RIV identification code

RIV/00216224:14740/20:00115752

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1128/mBio.00226-20

UT WoS

000531071300056

Keywords in English

RTX toxins; cell adhesion; clip-and-link; host-pathogen interactions; nuclear magnetic resonance

Abstract

V originále

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 epsilon-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.

Links

LM2018133, research and development project

Name: Český národní uzel Evropské infrastruktury pro translační medicínu (Acronym: EATRIS-ERIC-CZ)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

90127, large research infrastructures

Name: CIISB II

Citovat

KUBÁŇ, Vojtěch, P. MACEK, Jozef HRITZ, K. NECHVATALOVA, K. NEDBALCOVA, M. FALDYNA, Peter ŠEBO, Lukáš ŽÍDEK and L. BUMBA. Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins. MBIO. Washington, D.C.: American Society for Microbiology, 2020, vol. 11, No 2, p. 1-18. ISSN 2150-7511. Available from: https://dx.doi.org/10.1128/mBio.00226-20.

@article{1660918,
   author = {Kubáň, Vojtěch and Macek, P. and Hritz, Jozef and Nechvatalova, K. and Nedbalcova, K. and Faldyna, M. and Šebo, Peter and Žídek, Lukáš and Bumba, L.},
   article_location = {Washington, D.C.},
   article_number = {2},
   doi = {http://dx.doi.org/10.1128/mBio.00226-20},
   keywords = {RTX toxins; cell adhesion; clip-and-link; host-pathogen interactions; nuclear magnetic resonance},
   language = {eng},
   issn = {2150-7511},
   journal = {MBIO},
   title = {Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins},
   url = {https://mbio.asm.org/content/mbio/11/2/e00226-20.full.pdf},
   volume = {11},
   year = {2020}
}

TY  - JOUR
ID  - 1660918
AU  - Kubáň, Vojtěch - Macek, P. - Hritz, Jozef - Nechvatalova, K. - Nedbalcova, K. - Faldyna, M. - Šebo, Peter - Žídek, Lukáš - Bumba, L.
PY  - 2020
TI  - Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins
JF  - MBIO
VL  - 11
IS  - 2
SP  - 1-18
EP  - 1-18
PB  - American Society for Microbiology
SN  - 21507511
KW  - RTX toxins
KW  - cell adhesion
KW  - clip-and-link
KW  - host-pathogen interactions
KW  - nuclear magnetic resonance
UR  - https://mbio.asm.org/content/mbio/11/2/e00226-20.full.pdf
L2  - https://mbio.asm.org/content/mbio/11/2/e00226-20.full.pdf
N2  - 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 epsilon-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.
ER  -

KUBÁŇ, Vojtěch, P. MACEK, Jozef HRITZ, K. NECHVATALOVA, K. NEDBALCOVA, M. FALDYNA, Peter ŠEBO, Lukáš ŽÍDEK and L. BUMBA. Structural Basis of Ca2+-Dependent Self-Processing Activity of Repeat-in-Toxin Proteins. \textit{MBIO}. Washington, D.C.: American Society for Microbiology, 2020, vol.~11, No~2, p.~1-18. ISSN~2150-7511. Available from: https://dx.doi.org/10.1128/mBio.00226-20.

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