Imperial College London
Alternate

ProfessorDavidHolden

Faculty of Medicine, Department of Infectious Disease

Professor
 
 
 
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Contact

 

+44 (0)20 7594 3073d.holden

 
 
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Location

 

221Flowers buildingSouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Grabe:2016:10.1074/jbc.M116.752782,
author = {Grabe, GJ and Zhang, Y and Przydacz, M and Rolhion, N and Yang, Y and Pruneda, JN and Komander, D and Holden, DW and Hare, SA},
doi = {10.1074/jbc.M116.752782},
journal = {Journal of Biological Chemistry},
pages = {25853--25863},
title = {The Salmonella effector SpvD is a cysteine hydrolase with a serovar-specific polymorphism influencing catalytic activity, suppression of immune responses and bacterial virulence},
url = {http://dx.doi.org/10.1074/jbc.M116.752782},
volume = {291},
year = {2016}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - 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 C73, H162, and D182. 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.
AU  - Grabe,GJ
AU  - Zhang,Y
AU  - Przydacz,M
AU  - Rolhion,N
AU  - Yang,Y
AU  - Pruneda,JN
AU  - Komander,D
AU  - Holden,DW
AU  - Hare,SA
DO  - 10.1074/jbc.M116.752782
EP  - 25863
PY  - 2016///
SN  - 1083-351X
SP  - 25853
TI  - The Salmonella effector SpvD is a cysteine hydrolase with a serovar-specific polymorphism influencing catalytic activity, suppression of immune responses and bacterial virulence
T2  - Journal of Biological Chemistry
UR  - http://dx.doi.org/10.1074/jbc.M116.752782
UR  - http://www.ncbi.nlm.nih.gov/pubmed/27789710
UR  - http://hdl.handle.net/10044/1/42661
VL  - 291
ER  -
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