Antipeptide antibodies directed to the C-terminal part of ammodytoxin A react with the PLA2 subunit of crotoxin and neutralize its pharmacological activity.
Crotoxin and ammodytoxin A are snake venom neurotoxic phospholipases A2. Polyclonal antibodies against three synthetic peptides selected from the C-terminal part of the primary structure of ammodytoxin A were tested by ELISA for their interaction with crotoxin and its subunits, CA and CB. All three antipeptide antibodies reacted specifically with corresponding parts of ammodytoxin A and CB, either native or reduced. Conversely, polyclonal antibodies produced against ammodytoxin A and CB reacted strongly with all three peptides, suggesting that they constitute at least a part of natural epitopes in both proteins. All antipeptide antibodies reacted also with the corresponding peptides derived from CB by cyanogen bromide cleavage. The biological activity of the immune complexes was tested. No significant change in the enzymatic activity of CB, ammodytoxin A or crotoxin was observed with any of the three antipeptide antibodies. These antibodies were, however, able to protect mice against the lethal potency of CB and to prolong survival time of mice injected with crotoxin. These antipeptide antibodies were assayed in vitro for their protective effect against the action of CB or crotoxin on synaptosomes from Torpedo marmorata electric organ. They partly inhibited the acetylcholine release induced by both proteins. These results indicate that the C-terminal part of CB is likely to be involved in the pharmacological action of crotoxin.
Curin-Serbec
V
V
Unité des Venins, Institut Pasteur, Paris, France.
Délot
E
E
Faure
G
G
Saliou
B
B
Gubensek
F
F
Bon
C
C
Choumet
V
V
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000633:toxicity; D000997:Antivenins / Q000008:administration & dosage / Q000627:therapeutic use; D015153:Blotting, Western; D003198:Computer Simulation; D003439:Crotoxin / Q000737:chemistry / Q000276:immunology* / Q000633:toxicity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010741:Phospholipases A / Q000737:chemistry / Q000276:immunology*; D054467:Phospholipases A2; D011041:Poisoning / Q000628:therapy; D017386:Sequence Homology, Amino Acid; D013574:Synaptosomes / Q000187:drug effects; D014101:Torpedo; D014757:Viper Venoms / Q000737:chemistry / Q000276:immunology*
D000906:Antibodies; D000936:Antigen-Antibody Complex; D000997:Antivenins; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D014757:Viper Venoms; D003439:Crotoxin; C068117:ammodytoxin A; D010741:Phospholipases A; D054467:Phospholipases A2