Structure-function relationships in the neurotoxin Vipoxin from the venom of Vipera ammodytes meridionalis.
The neurotoxic complex Vipoxin is the lethal component of the venom of Vipera ammodytes meridionalis, the most toxic snake in Europe. It is a complex between a toxic phospholipase A2 (PLA2) and a non-toxic and catalytically inactive protein, stabilizing the enzyme and reducing the activity and toxicity. Structure-function relationships in this complex were studied by spectroscopic methods. A good correlation between the ionization behaviour and accessible surface area (ASA) of the tyrosyl residues was observed. In the toxic PLA2 subunit phenolic groups participate in H-bonding network that stabilizes the catalytically and pharmacologically active conformation. The tryptophan fluorescence decay of Vipoxin is well fitted by two exponentials with lifetimes of 0.1 (54%) and 2.5 (46%) ns. W20P, W31P and W31I are located in the interface between the two subunits and participate in hydrophobic interactions stabilizing the complex. Dissociation of the complex leads to a transition of the tryptophans from hydrophobic to hydrophilic environment, which influences mainly tau2. The longer lifetime is more sensitive to the polarity of the environment. Circular dichroism measurements demonstrate that the two components of the neurotoxin preserve their secondary structure after dissociation of the complex. The results of the spectroscopic studies are in accordance with a mechanism of blockade of transmission across the neuromuscular junctions of the breathing muscles by interaction of a dissociated toxic PLA2 with a membrane. The loss of toxicity is connected with slight changes in the secondary structure of PLA2. CD studies also show a substantial contribution of disulfide bonds to the stability of the neurotoxic complex and its components.
Georgieva
Dessislava Nikolova
DN
Institute of Organic Chemistry, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria.
Genov
Nicolay
N
Nikolov
Peter
P
Aleksiev
Boris
B
Rajashankar
Kanagalaghatta R
KR
Voelter
Wolfgang
W
Betzel
Christian
C
D000818:Animals; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D004220:Disulfides / Q000737:chemistry; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D009498:Neurotoxins / Q000737:chemistry* / Q000633:toxicity*; D010084:Oxidation-Reduction; D010741:Phospholipases A / Q000737:chemistry / Q000633:toxicity; D054467:Phospholipases A2; D021122:Protein Subunits; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D013329:Structure-Activity Relationship; D014364:Tryptophan / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry; D014757:Viper Venoms / Q000737:chemistry* / Q000633:toxicity*; D017819:Viperidae
D004220:Disulfides; D009498:Neurotoxins; D021122:Protein Subunits; D014757:Viper Venoms; D014443:Tyrosine; C033134:vipoxin; D014364:Tryptophan; D010741:Phospholipases A; D054467:Phospholipases A2