The Salmonella effector SpvD is a cysteine hydrolase with a serovar-specific polymorphism influencing catalytic activity, suppression of immune responses, and bacterial virulence

Grzegorz J. Grabe, Yue Zhang, Michal Przydacz, Nathalie Rolhion, Yi Yang, Jonathan Pruneda, David Komander, David W. Holden, Stephen A. Hare

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Many bacterial pathogens secrete virulence (effector) proteins that interfere with immune signaling in their host. SpvD is a Salmonella enterica effector protein that we previously demonstrated to negatively regulate the NF-κB signaling pathway and promote virulence of S. enterica serovar Typhimurium in mice. To shed light on the mechanistic basis for these observations, we determined the crystal structure of SpvD and show that it adopts a papain-like fold with a characteristic cysteine-histidine-aspartate catalytic triad comprising Cys-73, His-162, and Asp-182. SpvD possessed an in vitro deconjugative activity on aminoluciferin-linked peptide and protein substrates in vitro. A C73A mutation abolished SpvD activity, demonstrating that an intact catalytic triad is required for its function. Taken together, these results strongly suggest that SpvD is a cysteine protease. The amino acid sequence of SpvD is highly conserved across different S. enterica serovars, but residue 161, located close to the catalytic triad, is variable, with serovar Typhimurium SpvD having an arginine and serovar Enteritidis a glycine at this position. This variation affected hydrolytic activity of the enzyme on artificial substrates and can be explained by substrate accessibility to the active site. Interestingly, the SpvDG161 variant more potently inhibited NF-κB-mediated immune responses in cells in vitro and increased virulence of serovar Typhimurium in mice. In summary, our results explain the biochemical basis for the effect of virulence protein SpvD and demonstrate that a single amino acid polymorphism can affect the overall virulence of a bacterial pathogen in its host.

Original languageEnglish (US)
Pages (from-to)25853-25863
Number of pages11
JournalJournal of Biological Chemistry
Volume291
Issue number50
DOIs
StatePublished - Dec 9 2016
Externally publishedYes

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Salmonella
Hydrolases
Polymorphism
Cysteine
Virulence
Catalyst activity
Salmonella enterica
Pathogens
Proteins
Substrates
Amino Acids
Papain
Cysteine Proteases
Histidine
Aspartic Acid
Glycine
Arginine
Crystal structure
Amino Acid Sequence
Catalytic Domain

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The Salmonella effector SpvD is a cysteine hydrolase with a serovar-specific polymorphism influencing catalytic activity, suppression of immune responses, and bacterial virulence. / Grabe, Grzegorz J.; Zhang, Yue; Przydacz, Michal; Rolhion, Nathalie; Yang, Yi; Pruneda, Jonathan; Komander, David; Holden, David W.; Hare, Stephen A.

In: Journal of Biological Chemistry, Vol. 291, No. 50, 09.12.2016, p. 25853-25863.

Research output: Contribution to journalArticle

Grabe, Grzegorz J. ; Zhang, Yue ; Przydacz, Michal ; Rolhion, Nathalie ; Yang, Yi ; Pruneda, Jonathan ; Komander, David ; Holden, David W. ; Hare, Stephen A. / The Salmonella effector SpvD is a cysteine hydrolase with a serovar-specific polymorphism influencing catalytic activity, suppression of immune responses, and bacterial virulence. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 50. pp. 25853-25863.
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AU - Grabe, Grzegorz J.

AU - Zhang, Yue

AU - Przydacz, Michal

AU - Rolhion, Nathalie

AU - Yang, Yi

AU - Pruneda, Jonathan

AU - Komander, David

AU - Holden, David W.

AU - Hare, Stephen A.

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