dc.contributor.author |
Zheng, L |
en |
dc.contributor.author |
Khemlani, A |
en |
dc.contributor.author |
Lorenz, Natalie |
en |
dc.contributor.author |
Hughes, Jacelyn |
en |
dc.contributor.author |
Langley, Ries |
en |
dc.contributor.author |
Proft, Thomas |
en |
dc.date.accessioned |
2016-07-13T04:33:01Z |
en |
dc.date.issued |
2015-12 |
en |
dc.identifier.citation |
Journal of Biological Chemistry, 2015, 290 (52), pp. 31126 - 31137 |
en |
dc.identifier.issn |
0021-9258 |
en |
dc.identifier.uri |
http://hdl.handle.net/2292/29414 |
en |
dc.description.abstract |
Streptococcus pyogenes is an important human pathogen that causes a wide range of diseases. Using bioinformatics analysis of the complete S. pyogenes strain SF370 genome, we have identified a novel S. pyogenes virulence factor, which we termed streptococcal 5'-nucleotidase A (S5nA). A recombinant form of S5nA hydrolyzed AMP and ADP, but not ATP, to generate the immunomodulatory molecule adenosine. Michaelis-Menten kinetics revealed a Km of 169 μm and a Vmax of 7550 nmol/mg/min for the substrate AMP. Furthermore, recombinant S5nA acted synergistically with S. pyogenes nuclease A to generate macrophage-toxic deoxyadenosine from DNA. The enzyme showed optimal activity between pH 5 and pH 6.5 and between 37 and 47 °C. Like other 5'-nucleotidases, S5nA requires divalent cations and was active in the presence of Mg(2+), Ca(2+), or Mn(2+). However, Zn(2+) inhibited the enzymatic activity. Structural modeling combined with mutational analysis revealed a highly conserved catalytic dyad as well as conserved substrate and cation-binding sites. Recombinant S5nA significantly increased the survival of the non-pathogenic bacterium Lactococcus lactis during a human whole blood killing assay in a dose-dependent manner, suggesting a role as an S. pyogenes virulence factor. In conclusion, we have identified a novel S. pyogenes enzyme with 5'-nucleotidase activity and immune evasion properties. |
en |
dc.format.medium |
Print-Electronic |
en |
dc.language |
eng |
en |
dc.publisher |
American Society for Biochemistry and Molecular Biology |
en |
dc.relation.ispartofseries |
Journal of Biological Chemistry |
en |
dc.rights |
Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. Details obtained from http://www.sherpa.ac.uk/romeo/issn/0021-9258/ |
en |
dc.rights.uri |
https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm |
en |
dc.subject |
Macrophages |
en |
dc.subject |
Humans |
en |
dc.subject |
Lactococcus lactis |
en |
dc.subject |
Streptococcus pyogenes |
en |
dc.subject |
N-Glycosyl Hydrolases |
en |
dc.subject |
Bacterial Proteins |
en |
dc.subject |
Virulence Factors |
en |
dc.subject |
Blood Bactericidal Activity |
en |
dc.subject |
Microbial Viability |
en |
dc.subject |
Immune Evasion |
en |
dc.title |
Streptococcal 5'-Nucleotidase A (S5nA), a Novel Streptococcus pyogenes Virulence Factor That Facilitates Immune Evasion |
en |
dc.type |
Journal Article |
en |
dc.identifier.doi |
10.1074/jbc.M115.677443 |
en |
pubs.issue |
52 |
en |
pubs.begin-page |
31126 |
en |
pubs.volume |
290 |
en |
dc.description.version |
VoR - Version of Record |
en |
dc.rights.holder |
Copyright:
American Society for Biochemistry and Molecular Biology |
en |
dc.identifier.pmid |
26527680 |
en |
pubs.end-page |
31137 |
en |
dc.rights.accessrights |
http://purl.org/eprint/accessRights/OpenAccess |
en |
pubs.subtype |
Article |
en |
pubs.elements-id |
503168 |
en |
pubs.org-id |
Medical and Health Sciences |
en |
pubs.org-id |
Medical Sciences |
en |
pubs.org-id |
Molecular Medicine |
en |
pubs.org-id |
Science |
en |
pubs.org-id |
Science Research |
en |
pubs.org-id |
Maurice Wilkins Centre (2010-2014) |
en |
dc.identifier.eissn |
1083-351X |
en |
pubs.record-created-at-source-date |
2016-07-13 |
en |
pubs.dimensions-id |
26527680 |
en |