Proteomics of snake venoms from Elapidae and Viperidae families by multidimensional chromatographic methods

Abstract

Snake venoms contain a large number of biologically active substances and the venom components are very useful for pharmaceutical applications. Our goal is to separate and identify components of snake venoms in ten snake species from the Elapidae and Viperidae families using multidimensional chromatographic methods. The multidimensional chromatographic methods include reversed-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), lab-on-a-chip, two-dimensional electrophoresis (2-DE), and mass spectrometry. The venoms of eight snake species demonstrated major differences in hydrophobicity, molecular weight separations, and 2-DE protein distribution patterns. The 2-DE images showed major differences between families, within each family and even between the same species. Venoms of the Elapidae family showed many basic proteins with a wide range of molecular weights, while venoms of the Viperidae family showed wide ranges of pI and molecular weights, especially for Trimeresurus sp. The multidimensional chromatographic methods revealed specific differences in venom proteins intra-species as well as between species and families. We have isolated and identified proteins that may be unique for each species for further studies in the proteome of snake venoms and their potentially use in the pharmaceutical applications.

The Role of Conventional Two-Dimensional Electrophoresis (2DE) and Its Newer Applications in the Study of Snake Venoms

“…The Elapidae venoms, on the other hand, had profiles with protein spots at the molecular weight range of 15-20kDa (Fig.1). These results clearly elicited the differences of the patterns between the Elapidae and Viperidae venoms that have been so well documented in other literatures discussed above, thus proving the feasibility of 2DE as an ancillary taxonomic tool (Calvete et al, 2009;Nawarak et al, 2003). Apart from inter-family difference in venom compositions, the 2DE analysis of the eight Malaysian snake venoms also demonstrated the obvious pattern of train of spots due to post-translational modifications in venom proteins (Fig.…”

“…Hence, researchers who choose to utilize the technique are generally aware of the limitations and are usually able to adapt the complexity of the sample to the resolution power of the 2DE method by narrowing the study subject to one focus in order to reduce the sample complexity (Rabilloud et al, 2010). For instance, in a study done by Nawarak et al (Nawarak et al, 2003) to investigate the proteomes of a number of selected Elapidae and Viperidae venoms, the group first fractionated the crude venoms on RP-HPLC before subjecting only the major eluted peaks to 2DE analysis. In addition, the robustness of the 2DE technique has been tested thoroughly and the influence of the various parameters on the intra-laboratory reproducibility has been investigated (Choe & Lee, 2003), thus making the 2DE process a strong technique for proteome profile building and for subsequent deposition in databases to be accessed by researchers worldwide to be used for reference.…”

In Vitro Protease Inhibition, Modulation of PLA2 Activity and Protein Interaction Studies of Calotropis gigantea

“…A powerful post-synaptic neurotoxin and cardiotoxin present in Indian cobra's venom acts on the synaptic gaps of nerves, paralyzing the muscles, leading to failure of respiratory system or cardiac arrest. Enzymes present in the venom components like hyaluronidase cause lysis and increase the spread of the venom [18][19][20][21][22].…”

Electrophoretic Characterization of Venom Proteins of <em>Montivipera xanthina</em> (Gray, 1849) (Ophidia: Viperidae)

“…To date, a wide number of proteins and polypeptides belonging to 20 venom protein families have been identified and characterized from the venoms of several viperid snake species (Gitter et al, 1957(Gitter et al, , 1959Perkins et al, 1993;Perkins Tomer, 1995;Sanz et al, 2006, Mackessy, 2010. The predominant presence of enzymatic components, especially hydrolytic enzymes, in viperid venoms has been documented by many proteomic studies (Nawarak et al, 2003;Serrano et al, 2005;Sanz et al, 2006Sanz et al, , 2008Angulo et al, 2008;Mackessy, 2010).…”