Hydrogen Bonding and Equilibrium Protium−Deuterium Fractionation Factors in the Immunoglobulin G Binding Domain of Protein G†

Biochemistry ◽  
1999 ◽  
Vol 38 (13) ◽  
pp. 3918-3925 ◽  
Author(s):  
Devesh Khare ◽  
Patrick Alexander ◽  
John Orban
Keyword(s):  
Hydrogen Bonding ◽  
Immunoglobulin G ◽  
Binding Domain ◽  
Protein G ◽  
Fractionation Factors

scholarly journals S-Layer-Mediated Display of the Immunoglobulin G-Binding Domain of Streptococcal Protein G on the Surface of Caulobacter crescentus: Development of an Immunoactive Reagent

2007 ◽  
Vol 73 (10) ◽  
pp. 3245-3253 ◽  
Author(s):  
John F. Nomellini ◽  
Gillian Duncan ◽  
Irene R. Dorocicz ◽  
John Smit
Keyword(s):  
Immunoglobulin G ◽  
Tandem Repeats ◽  
Caulobacter Crescentus ◽  
Protein A ◽  
Cost Effective ◽  
Binding Domain ◽  
Protein G ◽  
Wild Type ◽  
Igg Binding ◽  
Streptococcal Protein G

ABSTRACT The immunoglobulin G (IgG)-binding streptococcal protein G is often used for immunoprecipitation or immunoadsorption-based assays, as it exhibits binding to a broader spectrum of host species IgG and IgG subclasses than the alternative, Staphylococcus aureus protein A. Caulobacter crescentus produces a hexagonally arranged paracrystalline protein surface layer (S-layer) composed of a single secreted protein, RsaA, that is notably tolerant of heterologous peptide insertions while maintaining the surface-attached crystalline character. Here, a protein G IgG-binding domain, GB1, was expressed as an insertion into full-length RsaA on the cell surface to produce densely packed immunoreactive particles. GB1 insertions at five separate sites were expressed, and all bound rabbit and goat IgG, but expression levels were reduced compared to those of wild-type RsaA and poor binding to mouse IgG was noted. To remedy this, we used the 20-amino-acid Muc1 peptide derived from human mucins as a spacer, since insertions of multiple tandem repeats were well tolerated for RsaA secretion and assembly. This strategy worked remarkably well, and recombinant RsaA proteins, containing up to three GB1 domains, surrounded by Muc1 peptides, not only were secreted and assembled but did so at wild-type levels. The ability to bind IgG (including mouse IgG) increased as GB1 units were added, and those with three GB1 domains bound twice as much rabbit IgG per cell as S. aureus cells (Pansorbin). The ability of recombinant protein G-Caulobacter cells to function as immunoactive reagents was assessed in an immunoprecipitation assay using a FLAG-tagged protein and anti-FLAG mouse monoclonal antibody; their performance was comparable to that of protein G-Sepharose beads. This work demonstrates the potential for using cells expressing recombinant RsaA/GB1 in immunoassays, especially considering that protein G-Caulobacter cells are more cost-effective than protein G beads and exhibit a broader species and IgG isotype binding range than protein A.

Effects of segment substitution on the structure and stability of immunoglobulin G binding domain of streptococcal protein G

Biopolymers ◽  
2005 ◽  
Vol 79 (1) ◽  
pp. 9-17
Author(s):  
Hai-Ning Du ◽  
Tie-Ying Zhang ◽  
Yong-Gang Chang ◽  
Dong-Hai Lin ◽  
Hong-Yu Hu
Keyword(s):  
Immunoglobulin G ◽  
Binding Domain ◽  
Protein G ◽  
Structure And Stability ◽  
Streptococcal Protein G ◽  
Segment Substitution

Kinetics of the Antibody Recognition Site in the Third IgG-Binding Domain of Protein G

2016 ◽  
Vol 128 (33) ◽  
pp. 9719-9722 ◽  
Author(s):  
Supriya Pratihar ◽  
T. Michael Sabo ◽  
David Ban ◽  
R. Bryn Fenwick ◽  
Stefan Becker ◽  
...  
Keyword(s):  
Binding Domain ◽  
Recognition Site ◽  
Protein G ◽  
Antibody Recognition ◽  
The Third ◽  
Igg Binding ◽  
Kinetics Of

The Third IgG-Binding Domain from Streptococcal Protein G

1994 ◽  
Vol 243 (5) ◽  
pp. 906-918 ◽  
Author(s):  
Jeremy P. Derrick ◽  
Dale B. Wigley
Keyword(s):  
Binding Domain ◽  
Protein G ◽  
The Third ◽  
Igg Binding ◽  
Streptococcal Protein G

Kinetics of the Antibody Recognition Site in the Third IgG-Binding Domain of Protein G

2016 ◽  
Vol 55 (33) ◽  
pp. 9567-9570 ◽  
Author(s):  
Supriya Pratihar ◽  
T. Michael Sabo ◽  
David Ban ◽  
R. Bryn Fenwick ◽  
Stefan Becker ◽  
...  
Keyword(s):  
Binding Domain ◽  
Recognition Site ◽  
Protein G ◽  
Antibody Recognition ◽  
The Third ◽  
Igg Binding ◽  
Kinetics Of
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