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Binding of complement inhibitor C4b-binding protein to a highly virulent S. pyogenes M1 strain is mediated by protein H and enhances adhesion to and invasion of endothelial cells.

Ermert, David LU ; Weckel, Antonin; Agarwal, Vaibhav LU ; Frick, Inga-Maria LU ; Björck, Lars LU and Blom, Anna LU (2013) In Journal of Biological Chemistry 288(45). p.32172-32183
Abstract
Streptococcus pyogenes AP1, a strain of the highly virulent M1 serotype, uses exclusively protein H to bind the complement inhibitor C4b-binding protein (C4BP). We found a strong correlation between the ability of AP1 and its isogenic mutants lacking protein H to inhibit opsonisation with complement C3b and binding of C4BP. C4BP bound to immobilized protein H or AP1 bacteria retained its cofactor activity for degradation of 125I-C4b. Further, C4b deposited from serum onto AP1 bacterial surfaces was processed into C4c/C4d fragments, which did not occur on strains unable to bind C4BP. Recombinant C4BP mutants, which (i) lack certain CCP domains, or (ii) have mutations in single aa as well as (iii) mutants with additional aa between different... (More)
Streptococcus pyogenes AP1, a strain of the highly virulent M1 serotype, uses exclusively protein H to bind the complement inhibitor C4b-binding protein (C4BP). We found a strong correlation between the ability of AP1 and its isogenic mutants lacking protein H to inhibit opsonisation with complement C3b and binding of C4BP. C4BP bound to immobilized protein H or AP1 bacteria retained its cofactor activity for degradation of 125I-C4b. Further, C4b deposited from serum onto AP1 bacterial surfaces was processed into C4c/C4d fragments, which did not occur on strains unable to bind C4BP. Recombinant C4BP mutants, which (i) lack certain CCP domains, or (ii) have mutations in single aa as well as (iii) mutants with additional aa between different CCP domains were used to determine that the binding is mainly mediated by a patch of positively charged amino acid residues at the interface of domains CCP1 and CCP2. Using recombinant protein H fragments, we narrowed down the binding site to the N-terminal domain A. With a peptide microarray, we identified one single 18 amino acid long peptide comprising residues 92-109, which specifically bound C4BP. Biacore was used to determine KD = 6x10-7 M between protein H and a single subunit of C4BP. C4BP binding also correlated with elevated levels of adhesion and invasion to endothelial cells. Taken together, we identified the molecular basis of C4BP-protein H interaction and found that it is not only important for decreased opsonisation but also for invasion of endothelial cells by Streptococcus pyogenes. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
288
issue
45
pages
32172 - 32183
publisher
ASBMB
external identifiers
  • wos:000328681700007
  • pmid:24064215
  • scopus:84887432633
ISSN
1083-351X
DOI
10.1074/jbc.M113.502955
language
English
LU publication?
yes
id
4392502f-917c-4467-a69b-afa3c2a7fd4e (old id 4065406)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/24064215?dopt=Abstract
date added to LUP
2013-10-03 22:03:58
date last changed
2019-09-17 01:13:13
@article{4392502f-917c-4467-a69b-afa3c2a7fd4e,
  abstract     = {Streptococcus pyogenes AP1, a strain of the highly virulent M1 serotype, uses exclusively protein H to bind the complement inhibitor C4b-binding protein (C4BP). We found a strong correlation between the ability of AP1 and its isogenic mutants lacking protein H to inhibit opsonisation with complement C3b and binding of C4BP. C4BP bound to immobilized protein H or AP1 bacteria retained its cofactor activity for degradation of 125I-C4b. Further, C4b deposited from serum onto AP1 bacterial surfaces was processed into C4c/C4d fragments, which did not occur on strains unable to bind C4BP. Recombinant C4BP mutants, which (i) lack certain CCP domains, or (ii) have mutations in single aa as well as (iii) mutants with additional aa between different CCP domains were used to determine that the binding is mainly mediated by a patch of positively charged amino acid residues at the interface of domains CCP1 and CCP2. Using recombinant protein H fragments, we narrowed down the binding site to the N-terminal domain A. With a peptide microarray, we identified one single 18 amino acid long peptide comprising residues 92-109, which specifically bound C4BP. Biacore was used to determine KD = 6x10-7 M between protein H and a single subunit of C4BP. C4BP binding also correlated with elevated levels of adhesion and invasion to endothelial cells. Taken together, we identified the molecular basis of C4BP-protein H interaction and found that it is not only important for decreased opsonisation but also for invasion of endothelial cells by Streptococcus pyogenes.},
  author       = {Ermert, David and Weckel, Antonin and Agarwal, Vaibhav and Frick, Inga-Maria and Björck, Lars and Blom, Anna},
  issn         = {1083-351X},
  language     = {eng},
  number       = {45},
  pages        = {32172--32183},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Binding of complement inhibitor C4b-binding protein to a highly virulent S. pyogenes M1 strain is mediated by protein H and enhances adhesion to and invasion of endothelial cells.},
  url          = {http://dx.doi.org/10.1074/jbc.M113.502955},
  volume       = {288},
  year         = {2013},
}