The
venom proteomics of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis, snakes of public health significance and the most poisonous reptiles in Europe, were analyzed by FPLC, 2-D electrophoresis, sequence analysis, and MS/MS. FPLC analysis showed the presence of
l-amino acid oxidase, monomeric and heterodimeric
phospholipases A2,
C-type lectin protein, and
proteinases in the
venom of V. a. ammodytes. Representatives of the same
protein families were found in the
venom of the other subspecies, V. a. meridionalis. N-terminally identical PLA2
neurotoxins were identified in both
venoms. Difference in the PLA2 compositions of the
venoms was also observed: a monomeric
protein with
phospholipase A2 activity, identical in the first 20
amino acid residues to the catalitically inactive acidic component of the heterodimeric PLA2 present in both
venoms, was found only in that of V. a. meridionalis. Probably, this
protein represents an intermediate form of the two components of the heterodimer. 2-D electrophoresis and MS/MS analysis showed that the two
venoms shared a number of
protein families: monomeric and heterodimeric Group II PLA2s,
serine proteinases, Group I, II, and III
metalloproteinases, l-
amino acid oxidases (LAAOs),
cysteine-rich secretory
proteins,
disintegrins, and
growth factors. Totally, 38
venom components of the V. a. ammodytes, belonging to 9
protein families, and 67 components of the V. a. meridionalis
venom belonging to 8
protein families were identified. The
venom proteome of V. a. ammodytes shows larger diversity of
proteins (139) in comparison to that of V. a. meridionalis (104
proteins). Most of the
proteins are homologues of known representatives of the respective
protein families. The
protein compositions explain clinical effects of the V. ammodytes
snakebites, such as difficulties in the breathing,
paralysis, apoptosis, cloting disorders,
hemorrhage, and tissue
necrosis. The lists of secreted
proteins by the two vipers can be used for further study of structure-function relationships in the toxins and for prediction and treatment of
snakebite consequences.