Interactions of the neurotoxin vipoxin in solution studied by dynamic light scattering.
The neurotoxin vipoxin is the lethal component of the venom of Vipera ammodytes meridionalis. It is a heterodimer of a basic toxic His-48 phospholipase A2 (PLA2) and an acidic nontoxic Gln-48 PLA2. The shape of the neurotoxin and its separated components in solution as well as their interactions with calcium, the brain phospholipid phosphatidylcholine, and two inhibitors, elaidoylamide and vitamin E, were investigated by dynamic light scattering. Calcium binding is connected with a conformational change in vipoxin observed as a change of the hydrodynamic shape from oblate ellipsoid to a shape closer to a sphere. The Ca2+-bound form of vipoxin, which is catalytically active, is more compact and symmetric than the calcium-free heterodimer. Similar changes were observed as a result of the Ca2+-binding to the two separated subunits. In the presence of aggregated phosphatidylcholine, the neurotoxic complex dissociates to subunits. It is supposed that only the toxic component binds to the substrate, and the other subunit, which plays a chaperone function, remains in solution. The inhibition of vipoxin with the synthetic inhibitor elaidoylamide and the natural compound vitamin E changes the shape of the toxin from oblate to prolate ellipsoid. The inhibited toxin is more asymmetric in comparison to the native one. Similar, but not so pronounced, effects were observed after the inhibition of the monomeric and homodimeric forms of the toxic His-48 PLA2. Circular dichroism measurements in the presence of urea, methylurea, and ethylurea indicate a strong hydrophobic stabilization of the neurotoxin. Hydrophobic interactions stabilize not only the folded regions but also the regions of intersubunit contacts.
Georgieva
Dessislava Nikolova
DN
Universitätsklinikum Hamburg-Eppendorf, Zentrum für Experimentelle Medizin, Institut für Biochemie und Molekularbiologie I, 22603 Hamburg, Germany.
Genov
Nicolay
N
Hristov
Krassimir
K
Dierks
Karsten
K
Betzel
Christian
C
D001665:Binding Sites; D002118:Calcium / Q000737:chemistry*; D002384:Catalysis; D003198:Computer Simulation; D057927:Hydrophobic and Hydrophilic Interactions; D007834:Lasers; D008958:Models, Molecular; D009498:Neurotoxins / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry*; D010783:Photometry / Q000379:methods*; D011485:Protein Binding; D011487:Protein Conformation; D012031:Refractometry / Q000379:methods*; D012542:Scattering, Radiation; D012996:Solutions; D014757:Viper Venoms / Q000037:antagonists & inhibitors / Q000737:chemistry*; D014810:Vitamin E / Q000737:chemistry*
D009498:Neurotoxins; D010713:Phosphatidylcholines; D012996:Solutions; D014757:Viper Venoms; D014810:Vitamin E; C033134:vipoxin; D002118:Calcium