A biomimetic Protein G affinity adsorbent: an Ugi ligand for immunoglobulins and Fab fragments based on the third IgG-binding domain of Protein G

2013 ◽  
Vol 26 (4) ◽  
pp. 190-200 ◽  
Author(s):  
Graziella El Khoury ◽  
Christopher R. Lowe
Keyword(s):  
Binding Domain ◽  
Protein G ◽  
The Third ◽  
Fab Fragments ◽  
Igg Binding ◽  
Affinity Adsorbent

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

scholarly journals ProCS15: A DFT-based chemical shift predictor for backbone and Cβ atoms in proteins

2015 ◽  
Author(s):  
Anders S Larsen ◽  
Lars A Bratholm ◽  
Anders S Christensen ◽  
Maher Jan Channir ◽  
Jan H. Jensen
Keyword(s):  
Hydrogen Bonding ◽  
Chemical Shift ◽  
Chemical Shifts ◽  
Structural Models ◽  
Protein G ◽  
Chemical Shielding ◽  
The Third ◽  
Entire Structure ◽  
Igg Binding ◽  
Structural Ensembles

We present ProCS15: A program that computes the isotropic chemical shielding values of backbone and C β atoms given a protein structure in less than a second. ProCS15 is based on around 2.35 million OPBE/6-31G(d,p)//PM6 calculations on tripeptides and small structural models of hydrogen-bonding. The ProCS15-predicted chemical shielding values are compared to experimentally measured chemical shifts for Ubiquitin and the third IgG-binding domain of Protein G through linear regression and yield RMSD values of up to 2.2, 0.7, and 4.8 ppm for carbon, hydrogen, and nitrogen atoms. These RMSD values are very similar to corresponding RMSD values computed using OPBE/6-31G(d,p) for the entire structure for each proteins. These maximum RMSD values can be reduced by using NMR-derived structural ensembles of Ubiquitin. For example, for the largest ensemble the largest RMSD values are 1.7, 0.5, and 3.5 ppm for carbon, hydrogen, and nitrogen. The corresponding RMSD values predicted by several empirical chemical shift predictors range between 0.7 - 1.1, 0.2 - 0.4, and 1.8 - 2.8 ppm for carbon, hydrogen, and nitrogen atoms, respectively.

Kinetic analysis of folding and unfolding the 56 amino acid IgG-binding domain of streptococcal protein G

Biochemistry ◽  
1992 ◽  
Vol 31 (32) ◽  
pp. 7243-7248 ◽  
Author(s):  
Patrick Alexander ◽  
John Orban ◽  
Philip Bryan
Keyword(s):  
Amino Acid ◽  
Kinetic Analysis ◽  
Binding Domain ◽  
Protein G ◽  
Igg Binding ◽  
Streptococcal Protein G

scholarly journals Equilibrium and pre-equilibrium fluorescence spectroscopic studies of the binding of a single-immunoglobulin-binding domain derived from protein G to the Fc fragment from human IgG1

1995 ◽  
Vol 310 (1) ◽  
pp. 177-184 ◽  
Author(s):  
K N Walker ◽  
S P Bottomley ◽  
A G Popplewell ◽  
B J Sutton ◽  
M G Gore
Keyword(s):  
Protein A ◽  
Spectroscopic Studies ◽  
Binding Domain ◽  
Protein G ◽  
Binding Interaction ◽  
Fc Fragment ◽  
Igg Binding ◽  
Immunoglobulin Binding Protein ◽  
Human Igg1 ◽  
Immunoglobulin Binding

A single-immunoglobulin-binding protein based upon the C2 domain of Protein G from Streptococcus has been shown to bind tightly to the Fc fragment of IgG1. The binding interaction results in a decrease in the fluorescence intensity from the sole Trp residue (Trp-48) in this domain. This spectral change has been used to monitor the binding interactions between the two proteins using equilibrium and pre-equilibrium fluorescence spectroscopy. Comparison of the data from the two techniques suggests that a conformational change occurs after the initial formation of the complex. Mutagenesis studies have shown that the Trp residue is important for binding and that replacement by a Phe residue is important for binding and that replacement by a Phe residue leads to a 300-fold decrease in the affinity for Fc gamma 1. Determination of the rate constants kon and koff at different values of pH between 4.0 and 9.0 suggest that variations in Kd are mediated predominantly by changes in kon. Competition experiments between SpG1 and a single-IgG-binding domain from Protein A from Staphylococcus aureus have been used to determine the affinity of the latter for Fc gamma 1.

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.

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