Proteomic Analyses of Agkistrodon contortrix contortrix Venom Using 2D Electrophoresis and MS Techniques

Abstract

Snake venom is a complex mixture of proteins and peptides which in the Viperidae is mainly hemotoxic. The diversity of these components causes the venom to be an extremely interesting object of study. Discovered components can be used in search for new pharmaceuticals used primarily in the treatment of diseases of the cardiovascular system. In order to determine the protein composition of the southern copperhead venom, we have used high resolution two dimensional electrophoresis and MALDI ToF/ToF MS-based identification. We have identified 10 groups of proteins present in the venom, of which phospholipase A2 and metalloprotease and serine proteases constitute the largest groups. For the first time presence of 5′-nucleotidase in venom was found in this group of snakes. Three peptides present in the venom were also identified. Two of them as bradykinin-potentiating agents and one as an inhibitor.

Comparison of Methods for Measuring Protein Concentration in Venom Samples

“…In the 2-D Quant Kit method, copper ions attach to the peptide bonds in the protein, so this method is independent of the amino acid composition of the proteins in the sample. This kit has been repeatedly used to measure the concentration of proteins in venom [28,39] and this is also the most commonly used method in our laboratory [10,34,35,40]. However, obtained results by 2-D Quant Kit and Bradford for A. contortrix venom show low variability, whereas for N. ashei venom, the differences are large.…”

“…However, obtained results by 2-D Quant Kit and Bradford for A. contortrix venom show low variability, whereas for N. ashei venom, the differences are large. This observation could explain why we have never managed to obtain well-separated and optimized gels for species of the genus Naja [40] and Dendroaspis, and we have had no problems with Viperids [10,34]. As the gel staining procedure involves the use of Coomassie Brillant Blue, which is also the dye in the Bradford method, the significant difference in the concentration values obtained from Bradford and 2-D Quant kit assays in the case of Naja ashei may be the reason why we observed gels that were not stained enough as if we added too few proteins.…”

Harnessing snake venom phospholipases A2 to novel approaches for overcoming antibiotic resistance

“…Generally, svPLA 2 s are among the main protein components of snake venoms (Bernardes et al, ; Boldrini‐Franca et al, ; de Oliveira et al, ). Proteomic and functional studies on snake venoms from Agkistrodon contortrix contortrix (Bocian et al, ) , Micrurus lemniscatus (Casais et al, ), and Agkistrodon piscivorus leucostoma (Lomonte et al, ) have revealed that PLA 2 s are the major component of these venom proteomes. In certain species of snakes, such as Bungarus multicinctus , svPLA 2 s correspond to up 60% of the proteome, covering different isoforms (Bocian et al, ).…”

“…5 0 NTs: 5 0 -nucleotidases, CTs: Cardiotoxins, PDEs: Phosphodiesterases, LAOs: L-aminooxidases, BPPs: Bradykininpotentiating peptides, SVSPs: Serine proteases, VEGFs: Snake venom vascular endothelial growth factors, DCs: Disintegrins, 3FTxs: Threefinger toxins, CRAMPs: Cathelin-related antimicrobial peptides, VHs: Venom hyaluronidases, SVMPS: Metalloproteinases, Lys49 PLA 2 s: Catalytically inactive phospholipases, CTLs: C-type lectin-like proteins, CRISPs: Cysteine-rich secretory proteins, Crots: Crotamines, Asp49 PLA 2 s: Catalytically active phospholipases, and pHpGs: Poly-Gly peptides 2009). Proteomic and functional studies on snake venoms from Agkistrodon contortrix contortrix (Bocian et al, 2016), Micrurus lemniscatus (Casais et al, 2016), and Agkistrodon piscivorus leucostoma (Lomonte et al, 2014) have revealed that PLA 2 s are the major component of these venom proteomes. In certain species of snakes, such as Bungarus multicinctus, svPLA 2 s correspond to up 60% of the proteome, covering different isoforms (Bocian et al, 2016).…”

Drugs Containing Snake Venom: History of Development, Nomenclature, Identification

“…В настоящее время в мировой практике идентификацию змеиных ядов проводят с помощью методов иммунологического анализа [17][18][19][20], массспектрометрии [9,11,[21][22][23][24], электрофореза [10,22,25,26] или сочетания этих методов (табл. 2).…”