Mechanistic insights into the neutralization of cytotoxic abrin by the monoclonal antibody D6F10.
Abrin, an A/B toxin obtained from the Abrus precatorius plant is extremely toxic and a potential bio-warfare agent. Till date there is no antidote or vaccine available against this toxin. The only known neutralizing monoclonal antibody against abrin, namely D6F10, has been shown to rescue the toxicity of abrin in cells as well as in mice. The present study focuses on mapping the epitopic region to understand the mechanism of neutralization of abrin by the antibody D6F10. Truncation and mutational analysis of abrin A chain revealed that the amino acids 74-123 of abrin A chain contain the core epitope and the residues Thr112, Gly114 and Arg118 are crucial for binding of the antibody. In silico analysis of the position of the mapped epitope indicated that it is present close to the active site cleft of abrin A chain. Thus, binding of the antibody near the active site blocks the enzymatic activity of abrin A chain, thereby rescuing inhibition of protein synthesis by the toxin in vitro. At 1∶10 molar concentration of abrin:antibody, the antibody D6F10 rescued cells from abrin-mediated inhibition of protein synthesis but did not prevent cell attachment of abrin. Further, internalization of the antibody bound to abrin was observed in cells by confocal microscopy. This is a novel finding which suggests that the antibody might function intracellularly and possibly explains the rescue of abrin's toxicity by the antibody in whole cells and animals. To our knowledge, this study is the first report on a neutralizing epitope for abrin and provides mechanistic insights into the poorly understood mode of action of anti-A chain antibodies against several toxins including ricin.
Department of Biochemistry, Indian Institute of Science, Bangalore, Karnataka, India.
Shradha
S
Bagaria
Devasena
D
Ponnalagu
Shveta
S
Bisht
Anjali A
AA
Karande
D000036:Abrin / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000276:immunology / Q000633:toxicity; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000737:chemistry* / Q000276:immunology; D000992:Antitoxins / Q000737:chemistry* / Q000276:immunology; D001665:Binding Sites; D020134:Catalytic Domain; D018604:Epitope Mapping; D006801:Humans; D062105:Molecular Docking Simulation; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D011485:Protein Binding / Q000276:immunology; D011487:Protein Conformation; D011817:Rabbits; D014118:Toxins, Biological / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000276:immunology / Q000633:toxicity
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000992:Antitoxins; D014118:Toxins, Biological; D000036:Abrin