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Affinity purification of human factor h on polypeptides derived from streptococcal m protein: enrichment of the y402 variant.

Nilsson, Rickard O LU ; Lannergård, Jonas LU ; Morgan, B Paul; Lindahl, Gunnar LU and Gustafsson, Mattias LU (2013) In PLoS ONE 8(11).
Abstract
Recent studies indicate that defective activity of complement factor H (FH) is associated with several human diseases, suggesting that pure FH may be used for therapy. Here, we describe a simple method to isolate human FH, based on the specific interaction between FH and the hypervariable region (HVR) of certain Streptococcus pyogenes M proteins. Special interest was focused on the FH polymorphism Y402H, which is associated with the common eye disease age-related macular degeneration (AMD) and has also been implicated in the binding to M protein. Using a fusion protein containing two copies of the M5-HVR, we found that the Y402 and H402 variants of FH could be efficiently purified by single-step affinity chromatography from human serum... (More)
Recent studies indicate that defective activity of complement factor H (FH) is associated with several human diseases, suggesting that pure FH may be used for therapy. Here, we describe a simple method to isolate human FH, based on the specific interaction between FH and the hypervariable region (HVR) of certain Streptococcus pyogenes M proteins. Special interest was focused on the FH polymorphism Y402H, which is associated with the common eye disease age-related macular degeneration (AMD) and has also been implicated in the binding to M protein. Using a fusion protein containing two copies of the M5-HVR, we found that the Y402 and H402 variants of FH could be efficiently purified by single-step affinity chromatography from human serum containing the corresponding protein. Different M proteins vary in their binding properties, and the M6 and M5 proteins, but not the M18 protein, showed selective binding of the FH Y402 variant. Accordingly, chromatography on a fusion protein derived from the M6-HVR allowed enrichment of the Y402 protein from serum containing both variants. Thus, the exquisite binding specificity of a bacterial protein can be exploited to develop a simple and robust procedure to purify FH and to enrich for the FH variant that protects against AMD. (Less)
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author
organization
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Contribution to journal
publication status
published
subject
in
PLoS ONE
volume
8
issue
11
publisher
Public Library of Science
external identifiers
  • wos:000327539800119
  • pmid:24278416
  • scopus:84896723274
ISSN
1932-6203
DOI
10.1371/journal.pone.0081303
language
English
LU publication?
yes
id
87477640-3236-4abe-89f7-415e64614351 (old id 4178900)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/24278416?dopt=Abstract
date added to LUP
2013-12-02 19:07:20
date last changed
2019-02-20 05:39:31
@article{87477640-3236-4abe-89f7-415e64614351,
  abstract     = {Recent studies indicate that defective activity of complement factor H (FH) is associated with several human diseases, suggesting that pure FH may be used for therapy. Here, we describe a simple method to isolate human FH, based on the specific interaction between FH and the hypervariable region (HVR) of certain Streptococcus pyogenes M proteins. Special interest was focused on the FH polymorphism Y402H, which is associated with the common eye disease age-related macular degeneration (AMD) and has also been implicated in the binding to M protein. Using a fusion protein containing two copies of the M5-HVR, we found that the Y402 and H402 variants of FH could be efficiently purified by single-step affinity chromatography from human serum containing the corresponding protein. Different M proteins vary in their binding properties, and the M6 and M5 proteins, but not the M18 protein, showed selective binding of the FH Y402 variant. Accordingly, chromatography on a fusion protein derived from the M6-HVR allowed enrichment of the Y402 protein from serum containing both variants. Thus, the exquisite binding specificity of a bacterial protein can be exploited to develop a simple and robust procedure to purify FH and to enrich for the FH variant that protects against AMD.},
  articleno    = {e81303},
  author       = {Nilsson, Rickard O and Lannergård, Jonas and Morgan, B Paul and Lindahl, Gunnar and Gustafsson, Mattias},
  issn         = {1932-6203},
  language     = {eng},
  number       = {11},
  publisher    = {Public Library of Science},
  series       = {PLoS ONE},
  title        = {Affinity purification of human factor h on polypeptides derived from streptococcal m protein: enrichment of the y402 variant.},
  url          = {http://dx.doi.org/10.1371/journal.pone.0081303},
  volume       = {8},
  year         = {2013},
}