title: Generation and expression of a minimal hybrid Ig-receptor formed between single domains from proteins L and G.
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.
authors.affiliation: Biomolecular Sciences Group, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.
authors.forename: Steven L
authors.initials: SL
authors.lastname: Harrison
authors.forename: Nicholas G
authors.initials: NG
authors.lastname: Housden
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Bottomley
authors.forename: Aimee J
authors.initials: AJ
authors.lastname: Cossins
authors.forename: Michael G
authors.initials: MG
authors.lastname: Gore
mesh_terms: D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D005822:Genetic Vectors; D007141:Immunoglobulin Fc Fragments / Q000276:immunology / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000276:immunology / Q000378:metabolism; D007136:Immunoglobulins / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D010462:Peptostreptococcus / Q000276:immunology; D015202:Protein Engineering; D011971:Receptors, Immunologic / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D013291:Streptococcus / Q000276:immunology
chemical_list: D001426:Bacterial Proteins; C055047:Ig L-binding protein, Peptostreptococcus; C034284:IgG Fc-binding protein, Streptococcus; D007141:Immunoglobulin Fc Fragments; D007145:Immunoglobulin kappa-Chains; D007136:Immunoglobulins; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins