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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4065406
- author
- Ermert, David
LU
; Weckel, Antonin
; Agarwal, Vaibhav
LU
; Frick, Inga-Maria
LU
; Björck, Lars
LU
and Blom, Anna
LU
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 288
- issue
- 45
- pages
- 32172 - 32183
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000328681700007
- pmid:24064215
- scopus:84887432633
- pmid:24064215
- 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
- 2016-04-01 10:11:50
- date last changed
- 2022-04-27 19:37:54
@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 = {{American Society for Biochemistry and Molecular Biology}},
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 = {{https://lup.lub.lu.se/search/files/1640417/4359996.pdf}},
doi = {{10.1074/jbc.M113.502955}},
volume = {{288}},
year = {{2013}},
}