title: Improving the anti-toxin abilities of the CMG2-Fc fusion protein with the aid of computational design.
abstract: CMG2-Fc is a fusion protein composed of the extracellular domain of capillary morphogenesis protein 2 (CMG2) and the Fc region of human immunoglobulin G; CMG2-Fc neutralizes anthrax toxin and offers protection against Bacillus anthracis challenge. To enhance the efficacy of CMG2-Fc against anthrax toxin, we attempted to engineer a CMG2-Fc with an improved affinity for PA. Using the automatic design algorithm FoldX and visual inspection, we devised two CMG2-Fc variants that introduce mutations in the CMG2 binding interface and improve the computationally assessed binding affinity for PA. An experimental affinity assay revealed that the two variants showed increased binding affinity, and in vitro and in vivo toxin neutralization testing indicated that one of these mutants (CMG2-Fc(E117Q)) has superior activity against anthrax toxin and was suitable for further development as a therapeutic agent for anthrax infections. This study shows that the computational design of the PA binding interface of CMG2 to obtain CMG2-Fc variants with improving anti-toxin abilities is viable. Our results demonstrate that computational design can be further applied to generate other CMG2-Fc mutants with greatly improved therapeutic efficacy.
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Yongyi
authors.initials: Y
authors.lastname: Xi
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Wu
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Gao
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Peng
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Hongxing
authors.initials: H
authors.lastname: Chen
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Huipeng
authors.initials: H
authors.lastname: Chen
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Xianwen
authors.initials: X
authors.lastname: Hu
authors.affiliation: Beijing Institute of Biotechnology, Beijing, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Yue
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics; D000992:Antitoxins / Q000737:chemistry / Q000235:genetics; D001408:Bacillus anthracis / Q000235:genetics / Q000472:pathogenicity; D001427:Bacterial Toxins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D003198:Computer Simulation; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000235:genetics; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D020125:Mutation, Missense; D018000:Receptors, Peptide / Q000737:chemistry / Q000235:genetics
chemical_list: C447150:ANTXR2 protein, human; D000942:Antigens, Bacterial; D000992:Antitoxins; D001427:Bacterial Toxins; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D018000:Receptors, Peptide; C030325:anthrax toxin
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