Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1953517 | Biochimie | 2006 | 9 Pages |
Abstract
R180, isolated from porcine brain cortex, is a high-affinity membrane receptor for ammodytoxin A (AtxA), a secreted phospholipase A2 (sPLA2) and presynaptically active neurotoxin from venom of the long-nosed viper (Vipera ammodytes ammodytes). As a member of the M-type sPLA2 receptors, present on the mammalian plasma membrane, R180 has been proposed to be responsible for one of the first events in the process of presynaptic neurotoxicity, the binding of the toxin to the nerve cell. To test this hypothesis, we prepared and analyzed three N-terminal fusion proteins of AtxA possessing a 12 or 5 amino acid residue peptide. The presence of such an additional “propeptide” prevented interaction of the toxin with the M-type receptor but not its lethality in mouse and neurotoxic effects on a mouse phrenic nerve-hemidiaphragm preparation. In addition, antibodies raised against the sPLA2-binding C-type lectin-like domain 5 of the M-type sPLA2 receptor were unable to abolish the neurotoxic action of AtxA on the neuromuscular preparation. The specific enymatic activities of the fusion AtxAs were two to three orders of magnitude lower from that of the wild type, yet resulting in a similar but less pronounced neurotoxic profile on the neuromuscular junction. This is in accordance with other data showing that a minimal enzymatic activity suffices for presynaptic toxicity of sPLA2s to occur. Our results indicate that the interaction of AtxA with the M-type sPLA2 receptor at the plasma membrane is not essential for presynaptic activity of the toxin. Interaction of AtxA with two intracellular proteins, calmodulin and the R25 receptor, was affected but not prevented by the presence of the N-terminal fusion peptides, implying that these proteins may play a role in the sPLA2 neurotoxicity.
Keywords
DABM-type phospholipase A2 receptorSecreted PLA2C-type lectin-like domainPOPGCTLDFABPsPLA2PLA2ATXABS1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol3,3′-diaminobenzidineVipera ammodytes ammodytesAmmodytoxinantibodiesphospholipase A2egg-yolk phosphatidylcholineSnake venomCAMNeurotoxicityNeuromuscularHorseradish peroxidaseFatty acid-binding proteinCalmodulin
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Authors
Petra Prijatelj, Nina Vardjan, Edward G. Rowan, Igor Križaj, Jože PungerÄar,