Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins

Lepesheva, Anna; Osičková, Adriana; Holubová, Jana; Jurnečka, David; Knoblochová, Šárka; Espinosa-Vinals, Carlos Angel; Bumba, Ladislav; Škopová, Karolína; Fišer, Radovan; Osička, Radim; Šebo, Peter; Mašín, Jiří

doi:https://dx.doi.org/10.1038/s41598-021-99112-3

Number of the records: 1

Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins

1.

SYSNO ASEP	0547497
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins
Author(s)	Lepesheva, Anna (MBU-M) Osičková, Adriana (MBU-M)_{RID, ORCID} Holubová, Jana (MBU-M)_{RID, ORCID} Jurnečka, David (MBU-M)_ORCID Knoblochová, Šárka (MBU-M) Espinosa-Vinals, Carlos Angel (MBU-M) Bumba, Ladislav (MBU-M)_{RID, ORCID} Škopová, Karolína (MBU-M) Fišer, Radovan (MBU-M)_{RID, ORCID} Osička, Radim (MBU-M)_{RID, ORCID} Šebo, Peter (MBU-M)_{RID, ORCID} Mašín, Jiří (MBU-M)_{RID, ORCID}
Article number	19814
Source Title	Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322 Roč. 11, č. 1 (2021)
Number of pages	16 s.
Language	eng - English
Country	DE - Germany
Keywords	adenylate-cyclase toxin ; escherichia-coli hemolysin ; bordetella-pertussis cyaa ; cell-invasive activity ; alpha-hemolysin ; membrane translocation ; complement receptor-3 ; fatty-acylation ; calcium ; binding
Subject RIV	CE - Biochemistry
OECD category	Biochemistry and molecular biology
R&D Projects	LM2018133 GA MŠMT - Ministry of Education, Youth and Sports (MEYS)
	GA19-12695S GA ČR - Czech Science Foundation (CSF)
	GX19-27630X GA ČR - Czech Science Foundation (CSF)
	GA19-04607S GA ČR - Czech Science Foundation (CSF)
Research Infrastructure	CIISB II - 90127 - Masarykova univerzita
Method of publishing	Open access
Institutional support	MBU-M - RVO:61388971
UT WOS	000706380800096
EID SCOPUS	85116409448
DOI	10.1038/s41598-021-99112-3
Annotation	Pore-forming repeats in toxins (RTX) are key virulence factors of many Gram-negative pathogens. We have recently shown that the aromatic side chain of the conserved tyrosine residue 940 within the acylated segment of the RTX adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) plays a key role in target cell membrane interaction of the toxin. Therefore, we used a truncated CyaA-derived RTX719 construct to analyze the impact of Y940 substitutions on functional folding of the acylated segment of CyaA. Size exclusion chromatography combined with CD spectroscopy revealed that replacement of the aromatic side chain of Y940 by the side chains of alanine or proline residues disrupted the calcium-dependent folding of RTX719 and led to self-aggregation of the otherwise soluble and monomeric protein. Intriguingly, corresponding alanine substitutions of the conserved Y642, Y643 and Y639 residues in the homologous RtxA, HlyA and ApxIA hemolysins from Kingella kingae, Escherichia coli and Actinobacillus pleuropneumoniae, affected the membrane insertion, pore-forming (hemolytic) and cytotoxic capacities of these toxins only marginally. Activities of these toxins were impaired only upon replacement of the conserved tyrosines by proline residues. It appears, hence, that the critical role of the aromatic side chain of the Y940 residue is highly specific for the functional folding of the acylated domain of CyaA and determines its capacity to penetrate target cell membrane.
Workplace	Institute of Microbiology
Contact	Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
Year of Publishing	2022
Electronic address	https://www.nature.com/articles/s41598-021-99112-3

Number of the records: 1