title: Novel high-affinity binders of human interferon gamma derived from albumin-binding domain of protein G.
abstract: Recombinant ligands derived from small protein scaffolds show promise as robust research and diagnostic reagents and next generation protein therapeutics. Here, we derived high-affinity binders of human interferon gamma (hIFNγ) from the three helix bundle scaffold of the albumin-binding domain (ABD) of protein G from Streptococcus G148. Computational interaction energy mapping, solvent accessibility assessment, and in silico alanine scanning identified 11 residues from the albumin-binding surface of ABD as suitable for randomization. A corresponding combinatorial ABD scaffold library was synthesized and screened for hIFNγ binders using in vitro ribosome display selection, to yield recombinant ligands that exhibited K(d) values for hIFNγ from 0.2 to 10 nM. Molecular modeling, computational docking onto hIFNγ, and in vitro competition for hIFNγ binding revealed that four of the best ABD-derived ligands shared a common binding surface on hIFNγ, which differed from the site of human IFNγ receptor 1 binding. Thus, these hIFNγ ligands provide a proof of concept for design of novel recombinant binding proteins derived from the ABD scaffold.
authors: Institute of Microbiology of the ASCR, v. v. i., Vídeňská 1083, 142 20 Prague, Czech Republic.
authors: Jawid N
authors: JN
authors: Ahmad
authors: Jingjing
authors: J
authors: Li
authors: Lada
authors: L
authors: Biedermannová
authors: Milan
authors: M
authors: Kuchař
authors: Hana
authors: H
authors: Sípová
authors: Alena
authors: A
authors: Semerádtová
authors: Jiří
authors: J
authors: Cerný
authors: Hana
authors: H
authors: Petroková
authors: Pavel
authors: P
authors: Mikulecký
authors: Jiří
authors: J
authors: Polínek
authors: Ondřej
authors: O
authors: Staněk
authors: Jiří
authors: J
authors: Vondrášek
authors: Jiří
authors: J
authors: Homola
authors: Jan
authors: J
authors: Malý
authors: Radim
authors: R
authors: Osička
authors: Peter
authors: P
authors: Sebo
authors: Petr
authors: P
authors: Malý
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D015723:Gene Library; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D008958:Models, Molecular; D009154:Mutation; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism*; D013291:Streptococcus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C034284:IgG Fc-binding protein, Streptococcus; D011994:Recombinant Proteins; D012709:Serum Albumin; D007371:Interferon-gamma