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Single-step purification of human C4b-binding protein (C4BP) by affinity chromatography on a peptide derived from a streptococcal surface protein.

Persson, Jenny J LU and Lindahl, Gunnar LU (2005) In Journal of Immunological Methods 297(1-2). p.83-95
Abstract
Many Gram-positive bacteria express surface proteins that bind human plasma proteins. These bacterial proteins, and derivatives of them, are of interest for analysis of bacterial pathogenesis and as immunochemical tools. Well-characterized examples include the IgG-binding reagents staphylococcal protein A and streptococcal protein G, and the recently described streptococcal IgA-binding peptide Sap. Here, we show that a peptide derived from the streptococcal M22 protein can be used for single-step affinity purification of the human complement regulator Cob-binding protein (C4BP). Binding of C4BP was strongly enhanced by dimerization of the peptide via a C-terminal cysteine residue not present in the intact M22 protein. The purified C4BP had... (More)
Many Gram-positive bacteria express surface proteins that bind human plasma proteins. These bacterial proteins, and derivatives of them, are of interest for analysis of bacterial pathogenesis and as immunochemical tools. Well-characterized examples include the IgG-binding reagents staphylococcal protein A and streptococcal protein G, and the recently described streptococcal IgA-binding peptide Sap. Here, we show that a peptide derived from the streptococcal M22 protein can be used for single-step affinity purification of the human complement regulator Cob-binding protein (C4BP). Binding of C4BP was strongly enhanced by dimerization of the peptide via a C-terminal cysteine residue not present in the intact M22 protein. The purified C4BP had the expected binding characteristics, and acted as a cofactor for factor I in the degradation of Cob. Passage of serum through a peptide column under non-saturating conditions resulted in binding of > 99.5% of serum C4BP, implying that such a column can be used to deplete serum of C4BP. These data indicate that the C4BP-binding peptide is a versatile tool that can be used for simple and rapid purification of biologically active human C4BP or for removal of C4BP from serum. (c) 2005 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
depletion, peptide, streptococcal M protein, protein purification, C4BP
in
Journal of Immunological Methods
volume
297
issue
1-2
pages
83 - 95
publisher
Elsevier
external identifiers
  • wos:000228301700008
  • scopus:15244360991
ISSN
1872-7905
DOI
10.1016/j.jim.2004.11.024
language
English
LU publication?
yes
id
9a24bc3e-0b3a-4e1b-a6ec-1c1bd301a3ea (old id 134862)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15777933&dopt=Abstract
date added to LUP
2007-07-12 13:07:07
date last changed
2017-01-01 07:18:47
@article{9a24bc3e-0b3a-4e1b-a6ec-1c1bd301a3ea,
  abstract     = {Many Gram-positive bacteria express surface proteins that bind human plasma proteins. These bacterial proteins, and derivatives of them, are of interest for analysis of bacterial pathogenesis and as immunochemical tools. Well-characterized examples include the IgG-binding reagents staphylococcal protein A and streptococcal protein G, and the recently described streptococcal IgA-binding peptide Sap. Here, we show that a peptide derived from the streptococcal M22 protein can be used for single-step affinity purification of the human complement regulator Cob-binding protein (C4BP). Binding of C4BP was strongly enhanced by dimerization of the peptide via a C-terminal cysteine residue not present in the intact M22 protein. The purified C4BP had the expected binding characteristics, and acted as a cofactor for factor I in the degradation of Cob. Passage of serum through a peptide column under non-saturating conditions resulted in binding of > 99.5% of serum C4BP, implying that such a column can be used to deplete serum of C4BP. These data indicate that the C4BP-binding peptide is a versatile tool that can be used for simple and rapid purification of biologically active human C4BP or for removal of C4BP from serum. (c) 2005 Elsevier B.V. All rights reserved.},
  author       = {Persson, Jenny J and Lindahl, Gunnar},
  issn         = {1872-7905},
  keyword      = {depletion,peptide,streptococcal M protein,protein purification,C4BP},
  language     = {eng},
  number       = {1-2},
  pages        = {83--95},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Single-step purification of human C4b-binding protein (C4BP) by affinity chromatography on a peptide derived from a streptococcal surface protein.},
  url          = {http://dx.doi.org/10.1016/j.jim.2004.11.024},
  volume       = {297},
  year         = {2005},
}