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Structure-function analysis of Heterodimer Formation, oligomerization, and receptor binding of the staphylococcus aureus bi-component toxin LukGH

Badarau, Adriana ; Rouha, Harald ; Malafa, Stefan ; Logan, Derek T. LU orcid ; Håkansson, Maria LU ; Stulik, Lukas ; Dolezilkova, Ivana ; Teubenbacher, Astrid ; Gross, Karin LU and Maierhofer, Barbara , et al. (2015) In Journal of Biological Chemistry 290(1). p.142-156
Abstract

The bi-component leukocidins of Staphylococcus aureus are important virulence factors that lyse human phagocytic cells and contribute to immune evasion. The γ-hemolysins (HlgAB and HlgCB) and Panton-Valentine leukocidin (PVL or LukSF) were shown to assemble from soluble subunits into membrane-bound oligomers on the surface of target cells, creating barrel-like pore structures that lead to cell lysis. LukGH is the most distantly related member of this toxin family, sharing only 30-40% amino acid sequence identity with the others. We observed that, unlike other leukocidin subunits, recombinant LukH and LukG had low solubility and were unable to bind to target cells, unless both components were present. Using biolayer interferometry and... (More)

The bi-component leukocidins of Staphylococcus aureus are important virulence factors that lyse human phagocytic cells and contribute to immune evasion. The γ-hemolysins (HlgAB and HlgCB) and Panton-Valentine leukocidin (PVL or LukSF) were shown to assemble from soluble subunits into membrane-bound oligomers on the surface of target cells, creating barrel-like pore structures that lead to cell lysis. LukGH is the most distantly related member of this toxin family, sharing only 30-40% amino acid sequence identity with the others. We observed that, unlike other leukocidin subunits, recombinant LukH and LukG had low solubility and were unable to bind to target cells, unless both components were present. Using biolayer interferometry and intrinsic tryptophan fluorescence we detected binding of LukH to LukG in solution with an affinity in the low nanomolar range and dynamic light scattering measurements confirmed formation of a heterodimer. We elucidated the structure of LukGH by x-ray crystallography at 2.8-A˚ resolution. This revealed an octameric structure that strongly resembles that reported for HlgAB, but with important structural differences. Structure guided mutagenesis studies demonstrated that three salt bridges, not found in other bi-component leukocidins, are essential for dimer formation in solution and receptor binding. We detected weak binding of LukH, but not LukG, to the cellular receptor CD11b by biolayer interferometry, suggesting that in common with other members of this toxin family, the S-component has the primary contact role with the receptor. Thesenewinsights provide the basis for novel strategies to counteract this powerful toxin and Staphylococcus aureus pathogenesis.

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publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
290
issue
1
pages
15 pages
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • pmid:25371205
  • scopus:84920531939
ISSN
0021-9258
DOI
10.1074/jbc.M114.598110
language
English
LU publication?
no
additional info
Publisher Copyright: © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
id
0c6050ea-fcd9-433f-9c6a-11697bb89d30
date added to LUP
2022-04-25 11:23:27
date last changed
2024-03-21 07:29:39
@article{0c6050ea-fcd9-433f-9c6a-11697bb89d30,
  abstract     = {{<p>The bi-component leukocidins of Staphylococcus aureus are important virulence factors that lyse human phagocytic cells and contribute to immune evasion. The γ-hemolysins (HlgAB and HlgCB) and Panton-Valentine leukocidin (PVL or LukSF) were shown to assemble from soluble subunits into membrane-bound oligomers on the surface of target cells, creating barrel-like pore structures that lead to cell lysis. LukGH is the most distantly related member of this toxin family, sharing only 30-40% amino acid sequence identity with the others. We observed that, unlike other leukocidin subunits, recombinant LukH and LukG had low solubility and were unable to bind to target cells, unless both components were present. Using biolayer interferometry and intrinsic tryptophan fluorescence we detected binding of LukH to LukG in solution with an affinity in the low nanomolar range and dynamic light scattering measurements confirmed formation of a heterodimer. We elucidated the structure of LukGH by x-ray crystallography at 2.8-A˚ resolution. This revealed an octameric structure that strongly resembles that reported for HlgAB, but with important structural differences. Structure guided mutagenesis studies demonstrated that three salt bridges, not found in other bi-component leukocidins, are essential for dimer formation in solution and receptor binding. We detected weak binding of LukH, but not LukG, to the cellular receptor CD11b by biolayer interferometry, suggesting that in common with other members of this toxin family, the S-component has the primary contact role with the receptor. Thesenewinsights provide the basis for novel strategies to counteract this powerful toxin and Staphylococcus aureus pathogenesis.</p>}},
  author       = {{Badarau, Adriana and Rouha, Harald and Malafa, Stefan and Logan, Derek T. and Håkansson, Maria and Stulik, Lukas and Dolezilkova, Ivana and Teubenbacher, Astrid and Gross, Karin and Maierhofer, Barbara and Weber, Susanne and Jägerhofer, Michaela and Hoffman, David and Nagy, Eszter}},
  issn         = {{0021-9258}},
  language     = {{eng}},
  month        = {{01}},
  number       = {{1}},
  pages        = {{142--156}},
  publisher    = {{American Society for Biochemistry and Molecular Biology}},
  series       = {{Journal of Biological Chemistry}},
  title        = {{Structure-function analysis of Heterodimer Formation, oligomerization, and receptor binding of the <i>staphylococcus aureus</i> bi-component toxin LukGH}},
  url          = {{http://dx.doi.org/10.1074/jbc.M114.598110}},
  doi          = {{10.1074/jbc.M114.598110}},
  volume       = {{290}},
  year         = {{2015}},
}