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Isolation and characterization of a 14-kDa albumin-binding fragment of streptococcal protein G

Sjöbring, U LU ; Falkenberg, C LU ; Nielsen, E ; Akerström, B LU and Björck, L LU (1988) In Journal of immunology 140(5). p.9-1595
Abstract

Protein G, a streptococcal cell wall protein, has separate binding sites for human albumin and IgG. Streptococci expressing protein G were treated with the bacteriolytic agent mutanolysin. Several IgG- and human serum albumin (HSA)-binding peptides were identified in the material thus solubilized and one of these, a 14-kDa peptide, was found to bind HSA but not IgG in Western blot experiments. This molecule was purified by affinity chromatography on Sepharose coupled with HSA followed by gel filtration on Sepharose 6B and a final affinity chromatography on IgG-Sepharose, by which low Mr W(15 to 20 kDa)IgG-binding peptides were removed. In different binding experiments the purified 14-kDa peptide bound exclusively HSA and the equilibrium... (More)

Protein G, a streptococcal cell wall protein, has separate binding sites for human albumin and IgG. Streptococci expressing protein G were treated with the bacteriolytic agent mutanolysin. Several IgG- and human serum albumin (HSA)-binding peptides were identified in the material thus solubilized and one of these, a 14-kDa peptide, was found to bind HSA but not IgG in Western blot experiments. This molecule was purified by affinity chromatography on Sepharose coupled with HSA followed by gel filtration on Sepharose 6B and a final affinity chromatography on IgG-Sepharose, by which low Mr W(15 to 20 kDa)IgG-binding peptides were removed. In different binding experiments the purified 14-kDa peptide bound exclusively HSA and the equilibrium constant between the peptide and HSA was determined to be 3.4 X 10(8) M-1. The relation between the 14-kDa molecule and protein G was studied by analyzing the N-terminal amino acid sequence of the peptide and comparing it with the previously determined protein G sequence. The 40 N-terminal amino acids were found to be identical with an amino acid sequence starting at position 62 in the protein G molecule. These and previous data enabled us to locate the albumin binding to the repetitively arranged domains in the N-terminal half of the protein G molecule.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Bacterial Proteins/isolation & purification, Carrier Proteins/isolation & purification, Humans, Immunoglobulin G/metabolism, Molecular Weight, Peptide Fragments/isolation & purification, Receptors, Albumin, Receptors, Cell Surface/analysis, Serum Albumin/immunology, Streptococcus/analysis
in
Journal of immunology
volume
140
issue
5
pages
9 - 1595
publisher
American Association of Immunologists
external identifiers
  • scopus:0023906698
  • pmid:2831269
ISSN
0022-1767
language
English
LU publication?
yes
id
fa88df90-e1b7-4efc-85cf-7f77e68c9b9c
alternative location
https://www.jimmunol.org/content/140/5/1595.long
date added to LUP
2019-05-22 10:30:35
date last changed
2024-01-01 06:53:19
@article{fa88df90-e1b7-4efc-85cf-7f77e68c9b9c,
  abstract     = {{<p>Protein G, a streptococcal cell wall protein, has separate binding sites for human albumin and IgG. Streptococci expressing protein G were treated with the bacteriolytic agent mutanolysin. Several IgG- and human serum albumin (HSA)-binding peptides were identified in the material thus solubilized and one of these, a 14-kDa peptide, was found to bind HSA but not IgG in Western blot experiments. This molecule was purified by affinity chromatography on Sepharose coupled with HSA followed by gel filtration on Sepharose 6B and a final affinity chromatography on IgG-Sepharose, by which low Mr W(15 to 20 kDa)IgG-binding peptides were removed. In different binding experiments the purified 14-kDa peptide bound exclusively HSA and the equilibrium constant between the peptide and HSA was determined to be 3.4 X 10(8) M-1. The relation between the 14-kDa molecule and protein G was studied by analyzing the N-terminal amino acid sequence of the peptide and comparing it with the previously determined protein G sequence. The 40 N-terminal amino acids were found to be identical with an amino acid sequence starting at position 62 in the protein G molecule. These and previous data enabled us to locate the albumin binding to the repetitively arranged domains in the N-terminal half of the protein G molecule.</p>}},
  author       = {{Sjöbring, U and Falkenberg, C and Nielsen, E and Akerström, B and Björck, L}},
  issn         = {{0022-1767}},
  keywords     = {{Bacterial Proteins/isolation & purification; Carrier Proteins/isolation & purification; Humans; Immunoglobulin G/metabolism; Molecular Weight; Peptide Fragments/isolation & purification; Receptors, Albumin; Receptors, Cell Surface/analysis; Serum Albumin/immunology; Streptococcus/analysis}},
  language     = {{eng}},
  month        = {{03}},
  number       = {{5}},
  pages        = {{9--1595}},
  publisher    = {{American Association of Immunologists}},
  series       = {{Journal of immunology}},
  title        = {{Isolation and characterization of a 14-kDa albumin-binding fragment of streptococcal protein G}},
  url          = {{https://www.jimmunol.org/content/140/5/1595.long}},
  volume       = {{140}},
  year         = {{1988}},
}