Spectroscopic properties and stability of the neurotoxic complex. Vipoxin and its components.
The neurotoxin Vipoxin from the venom of Vipera ammodytes meridionalis is a complex between a toxic basic phospholipase A2 (PLA2) and a non-toxic acidic protein inhibitor (Inh). Tryptophan fluorescence parameters are determined for the complex and for its components. Iodide, caesium and acrylamide are not efficient quenchers of the Vipoxin indole emission. Increased accessibilities of tryptophans to ionic and neutral quenchers are found after the dissociation of the complex. Trp 20 and Trp 31 became more 'exposed' in the separated individuals proteins. The indole rings of the complex are located in a positively charged environment. Inspection of the Vipoxin X-ray model showed that the three tryptophyl side chains are located in the interface region between the enzyme and the inhibitor and are completely 'exposed' in the separated components of the complex. In Vipoxin an efficient 'interchain' energy transfer between tyrosyl and tryptophyl residues from different polypeptide chains occurs. Static quenching with acrylamide is also detected in PLA2 and Inh. The free energy changes deltaG D for the unfolding reactions of Vipoxin, PLA2 and Inh are determined in circular dichroism spectroscopy. The complex formation between the toxic PLA2 and the inhibitor increases deltaG HD2O to 23.5 kJ mol-1.
Institute of Organic Chemistry, Bulgarian Academy of Sciences, Sofia, Bulgaria.
N
N
Genov
P
P
Dolashka
B
B
Aleksiev
I
I
Mancheva
K R
KR
Rajashankar
C
C
Betzel
D020106:Acrylamide; D000178:Acrylamides / Q000494:pharmacology; D004355:Drug Stability; D004735:Energy Transfer; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000378:metabolism; D019791:Guanidine; D007700:Kinetics; D008958:Models, Molecular; D009498:Neurotoxins / Q000737:chemistry*; D010741:Phospholipases A / Q000737:chemistry* / Q000378:metabolism; D054467:Phospholipases A2; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence / Q000379:methods; D014364:Tryptophan; D014443:Tyrosine; D014757:Viper Venoms / Q000737:chemistry* / Q000378:metabolism
D000178:Acrylamides; D004791:Enzyme Inhibitors; D009498:Neurotoxins; D014757:Viper Venoms; D020106:Acrylamide; D014443:Tyrosine; C033134:vipoxin; D014364:Tryptophan; D010741:Phospholipases A; D054467:Phospholipases A2; D019791:Guanidine