Protein structure of the venom in nine species of snake: from bio-compounds to possible healing agents.

R.T Cristina,O.M Boldura,C.I Hulea,C Tulcan,E Dumitrescu,F Muselin,E Alexa,I Radulov,R Kocsis

doi:10.1590/1414-431x20199001

Abstract

Due to its various structures in bio-compounds, snake venom is the indisputable result of evolutionary stages of molecules with an increasingly complex structure, high specificity, and of great importance for medicine because of their potential. The present study proposed an underpinning examination of venom composition from nine species of venomous snakes using a useful and replicable methodology. The objective was the extension of the evaluation of protein fractions in the field up to 230 kDa to permit possible identification of some fractions that are insufficiently studied. The gel capillary electrophoresis method on the chip was performed using an Agilent 2100 bioassay with the 80 and 230-LabChip Protein kits. Interpretation of electrophoresis was performed using the Protein 2100 expert (Agilent) test software as follows: a) Protein 80 (peak size scale): 1.60, 3.5, 6.50, 15.00, 28.00, 46.00, 63.00, 95.00 kDa; b) Protein 230 (peak size scale): 4.50, 7.00, 15.00, 28.00, 46.00, 63.00, 95.00, 150.00, 240.00 kDa. The screening revealed the presence of compounds with a molecular weight greater than 80 kDa, in the case of Vipera aspis and Vipera xantina palestinae. For V. aspis, a 125 kDa molecular weight pro-coagulant protein was identified, known as being involved in the reduction of plasma clotting time without any direct activity in the fibrinogen coagulation process. The samples examined on the Protein 230-LabChip electrophoresis chip can be considered as a novelty with possible uses in medicine, requiring further approaches by advanced proteomics techniques to confirm the intimate structural features and biological properties of snake venoms.

Highlights

Due to the various structures of bio-compounds, snake venoms are the indisputable result of evolutionary stages for molecules with an increasingly complex structure and high specificity [1,2,3]
Determination of protein content The total protein content of the samples was determined according to the known classical methodology originally provided by Lowry in 1951, which was adapted for venom by Dutta et al [27], the calibration curve being prepared with bovine serum albumin (BSA) (Figure 1)
The Vipera toxins contain a representative of the heterodimeric PLA2 group, identified in the 13–14 kDa molecular mass interval with an occurrence of 15.4% for Vipera xantina palestinae; it was firstly described by Ovadia [34] and confirmed by other authors [35,36,37]

Summary

Introduction

Due to the various structures of bio-compounds (e.g., peptides, toxins, enzymes, up to 100 proteins, and different isoforms), snake venoms are the indisputable result of evolutionary stages for molecules with an increasingly complex structure and high specificity [1,2,3].This high protein amino acid abundance, generally possessing specific enzymatic and polypeptide characteristics, is often different from one snake species to another, but authors generally agree that these assemblies can be classified into several common chemical families like: phosphodiesterases [4], acetylcholinesterases [5], phospholipases [6,7], proteases (serine and metalloproteases) [8,9,10], disintegrins [11,12], as well as the so-called ‘‘threefinger toxins’’ tri-toxins: neuro-cardio-hemodyotoxins [13]. Due to the various structures of bio-compounds (e.g., peptides, toxins, enzymes, up to 100 proteins, and different isoforms), snake venoms are the indisputable result of evolutionary stages for molecules with an increasingly complex structure and high specificity [1,2,3]. This large blend of biochemical molecules present in snake venoms makes these various and complex structures attractive to the investigation of new therapeutic resources [14,15].

Results

Conclusion