Ammodytin L, an inactive phospholipase A2 homologue with myotoxicity in mice, binds to the presynaptic acceptor of the beta-neurotoxic ammodytoxin C in Torpedo: an indication for a phospholipase A2 activity-independent mechanism of action of beta-neurotoxins in fish?
A Ser48 phospholipase A2-homologue, ammodytin L, which is myotoxic in mammals and devoid of any phospholipase A2 activity, completely inhibits the specific binding of the neurotoxic phospholipase A2, ammodytoxin C, to fish presynaptic membranes from Torpedo marmorata electric organ. In cross-linking experiments, 125I-ammodytin L labels the same membrane proteins as 125I-ammodytoxin C (70, 38.5-57.4 and 19.7 kDa). The formation of these adducts is completely prevented by the presence of ammodytoxin C but not of a non-toxic phospholipase A2, ammodytin I2. A chimeric phospholipase A2, constructed by associating the N-terminal half of ammodytoxin to the C-terminal half of ammodytin L, possesses a low, but significant phospholipase A2 activity, however it is not toxic to mice, probably due to abolition of the specific neuronal acceptor binding in mammals. Nevertheless, the chimeric phospholipase A2 is able to interact with the ammodytoxin acceptor in Torpedo marmorata electric organ. The existence of neuronal acceptors for ammodytin L and for the chimeric phospholipase A2 suggests that they may act as neurotoxins in fish. As ammodytin L does not possess any enzymatic activity it, therefore, appears to be an excellent tool to investigate the mechanism of action of beta-neurotoxins independently of their phospholipase A2 activity.
Pungercar
J
J
Department of Biochemistry and Molecular Biology, Jozef Stefan Institute, Jamova, Slovenia.
Vucemilo
N
N
Faure
G
G
Bon
C
C
Verheij
H M
HM
Gubensek
F
F
Krizaj
I
I
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D017931:DNA Primers / Q000235:genetics; D004557:Electric Organ / Q000378:metabolism; D005399:Fishes; D054501:Group II Phospholipases A2; D066298:In Vitro Techniques; D051379:Mice; D009469:Neuromuscular Junction / Q000187:drug effects; D009498:Neurotoxins / Q000378:metabolism* / Q000633:toxicity*; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*; D054467:Phospholipases A2; D016133:Polymerase Chain Reaction; D017661:Receptors, Presynaptic / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D014101:Torpedo; D014757:Viper Venoms / Q000235:genetics / Q000378:metabolism* / Q000633:toxicity*
D017931:DNA Primers; D009498:Neurotoxins; D017661:Receptors, Presynaptic; D011993:Recombinant Fusion Proteins; D014757:Viper Venoms; C065419:ammodytin L; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054467:Phospholipases A2; C060161:ammodytoxin C