Protein profile analysis of Malaysian snake venoms by two-dimensional gel electrophoresis

Abstract

Snake venoms comprise a highly complex mixture of proteins, which requires for their characterization the use of versatile two-dimensional electrophoresis techniques. In the present study, venoms obtained from eight snakes (Ophiophagus hannah, Naja kaouthia, Naja sumatrana, Bungarus fasciatus, Trimeresurus sumatranus, Tropidolaemus wagleri, Enhydrina schistosa and Calloselasma rhodostoma) commonly found in Malaysia were separated based on two independent properties, isoelectric point (pI) and molecular weight (MW). Many differences in snake venoms at the inter-family, inter-subfamily, inter-genus and inter-species levels were revealed. Notably, proteins from individuals of the Viperidae family -Trimeresurus sumatranus, Tropidolaemus wagleri and Calloselasma rhodostoma -were found to be numerous and scattered by the two-dimensional gel electrophoresis (2DE) specifically in regions between 37 and 100 kDa compared to the Elapidae venom proteins. The latter were clustered at the basic and lower molecular mass region (less than 20 kDa). Trains of spots were commonly observed, indicating that these proteins may be derived from post-translational modifications. Ophiophagus hannah (Elapidae) revealed a great amount of protein spots in the higher molecular mass range when compared to Enhydrina schistosa, Naja kaouthia, Naja sumatrana and Bungarus fasciatus. Overall 2DE showed large differences in the venom profile of each species, which might be employed as an ancillary tool to the identification of venomous snake species.

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

“…With high probability, it can be assumed that the proteins in this venom are highly post-translationally modified, as shown by clearly visible spots trains in gels (Figure 1 and Figure 2). This phenomenon is characteristic for Viperidae family and was described already several times [13,16]. …”

“…However, the advantage of this protocol is mostly visible in the class of small proteins and peptides [21]. Furthermore, 2D electrophoresis allows the observation of post-translational modifications, which can be crucial for the immunogenicity of proteins [13]. However, in our opinion the observed differences arise from analyzed venom composition and not the analytical techniques used.…”

“…This technique can be used to compare proteomes of different species [13], investigate the influence of factors like sex [14], the area of occurrence [15], or to determine whether post-translational modifications occurred in proteins [16]. The combination of 2DE and Western blot helps in clinical pathology of snake bites and antitoxins mechanisms studies [17].…”

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

“…Arrows indicate trains of spots due to post-translational modifications. Only two out of the eight 2DE venom profiles are shown here (Vejayan et al, 2010).…”

The concept of two-dimensional electrophoresis-guided purification proven by isolation of rhodocetin from Calloselasma rhodostoma (Malayan pit viper)

“…Protein identification on 2DE profiles and between-gel comparisons allow investigation into snake venom variations that have applications in basic venom research and envenomation management (4). Two-dimensional gel electrophoresis profiles and their analyses have also emerged as new tools for taxonomic studies by allowing investigation into interspecies and intraspecies variations (5,6). It can also be employed in selecting donor snakes for antivenom production, as well as in quality control by detecting degradation of venom components in the preparation of venom samples for immunization (4,7).…”