Abstract
The Ig-binding properties of protein L from Peptostreptococcus magnus and protein G from Streptococcus have been successfully combined through the construction of a novel hybrid protein, consisting of a single Ig-binding domain from each protein. The biophysical and biochemical properties of this construct have been characterized through equilibrium and pre-equilibrium fluorescence spectroscopy, circular dichroism, isothermal titration calorimetry, affinity chromatography, and conformational stability studies using a chemical denaturant in order to examine the structure and availability of ligand binding sites in each domain. These studies show that despite the small size of the protein (Mw=16.5 kDa) each domain behaves in an independent manner with respect to the binding characteristics of the same domain in isolation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Motifs
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Bacterial Proteins / chemistry
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Bacterial Proteins / immunology
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Bacterial Proteins / metabolism*
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Base Sequence
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Binding Sites
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Genetic Vectors
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Immunoglobulin Fc Fragments / immunology
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Immunoglobulin Fc Fragments / metabolism
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Immunoglobulin kappa-Chains / immunology
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Immunoglobulin kappa-Chains / metabolism
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Immunoglobulins / immunology
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Immunoglobulins / metabolism*
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Molecular Sequence Data
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Peptostreptococcus / immunology
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Protein Engineering*
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Receptors, Immunologic / chemistry
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Receptors, Immunologic / immunology
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Receptors, Immunologic / metabolism*
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / isolation & purification
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Recombinant Fusion Proteins / metabolism
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Streptococcus / immunology
Substances
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Bacterial Proteins
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Ig L-binding protein, Peptostreptococcus
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IgG Fc-binding protein, Streptococcus
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Immunoglobulin Fc Fragments
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Immunoglobulin kappa-Chains
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Immunoglobulins
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Receptors, Immunologic
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Recombinant Fusion Proteins