Improving the anti-toxin abilities of the CMG2-Fc fusion protein with the aid of computational design

PLoS One. 2014 Aug 7;9(8):e104674. doi: 10.1371/journal.pone.0104674. eCollection 2014.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / chemistry
  • Antigens, Bacterial / genetics
  • Antitoxins* / chemistry
  • Antitoxins* / genetics
  • Bacillus anthracis / genetics
  • Bacillus anthracis / pathogenicity
  • Bacterial Toxins / antagonists & inhibitors
  • Bacterial Toxins / chemistry
  • Bacterial Toxins / genetics
  • Computer Simulation*
  • Humans
  • Immunoglobulin Fc Fragments* / chemistry
  • Immunoglobulin Fc Fragments* / genetics
  • Immunoglobulin G* / chemistry
  • Immunoglobulin G* / genetics
  • Models, Molecular*
  • Mutation, Missense
  • Receptors, Peptide* / chemistry
  • Receptors, Peptide* / genetics

Substances

  • ANTXR2 protein, human
  • Antigens, Bacterial
  • Antitoxins
  • Bacterial Toxins
  • Immunoglobulin Fc Fragments
  • Immunoglobulin G
  • Receptors, Peptide
  • anthrax toxin

Grants and funding

This work was supported in part by grants from the National Science Foundation of China (No. 3090592 & 81202445) and the Chinese National Science and Technology Major Project (Nos. 2012ZX09102301-001 & 2011ZX09102-001-30). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.