Strategies in ‘snake venomics’ aiming at an integrative view of compositional, functional, and immunological characteristics of venoms

Abstract

This work offers a general overview on the evolving strategies for the proteomic analysis of snake venoms, and discusses how these may be combined through diverse experimental approaches with the goal of achieving a more comprehensive knowledge on the compositional, toxic, and immunological characteristics of venoms. Some recent developments in this field are summarized, highlighting how strategies have evolved from the mere cataloguing of venom components (proteomics/venomics), to a broader exploration of their immunological (antivenomics) and functional (toxicovenomics) characteristics. Altogether, the combination of these complementary strategies is helping to build a wider, more integrative view of the life-threatening protein cocktails produced by venomous snakes, responsible for thousands of deaths every year.

Intact protein mass spectrometry reveals intraspecies variations in venom composition of a local population of Vipera kaznakovi in Northeastern Turkey

“…Most of these studies applied the so called 'bottomup' proteomic approaches, whereby intact proteins are typically digested with trypsin before tandem mass spectrometry analysis. Many workflows perform venom decomplexation prior to digestion, either by liquid chromatography (LC) or gel electrophoresis, or a combination of both [17]. The direct, in-solution digestion, or so called 'shotgun proteomics', allows for a fast qualitative overview, but suffers from a less quantitative breakdown of snake venom composition [17,18].…”

“…Many workflows perform venom decomplexation prior to digestion, either by liquid chromatography (LC) or gel electrophoresis, or a combination of both [17]. The direct, in-solution digestion, or so called 'shotgun proteomics', allows for a fast qualitative overview, but suffers from a less quantitative breakdown of snake venom composition [17,18]. For example, in shotgun experiments, the problem of protein inference often does not permit the differentiation of the numerous toxin isoforms present in venom [19].…”

Extended snake venomics by top-down in-source decay: Investigating the newly discovered Anatolian Meadow viper subspecies,Vipera anatolica senliki

“…6 Since then, the venomics community experienced a rapid expansion into different venomic applications and a multitude of proteome studies for a wide variety of snake families have been published. 7, 8 Today, decomplexation of snake venom proteomes can be achieved by several bottom-up protocols, combining multidimensional separation methods. 7, 9 However, bottom-up proteomics suffers from classical proteolytic drawbacks, a step that often obscures the differentiation of toxin proteoforms and prevents the identification of post-translational modifications (PTMs).…”

“…7, 8 Today, decomplexation of snake venom proteomes can be achieved by several bottom-up protocols, combining multidimensional separation methods. 7, 9 However, bottom-up proteomics suffers from classical proteolytic drawbacks, a step that often obscures the differentiation of toxin proteoforms and prevents the identification of post-translational modifications (PTMs). 10–13 In this context, the recently introduced top-down protocol for snake venoms provide remedy by directly analyzing intact proteins and peptides out of crude venoms using high-resolution tandem mass spectrometry.…”

Toxin-centric development approach for next-generation antivenoms

“…However, they suffer from the drawback of not being compatible with the human immune system due to their heterologous nature (Laustsen et al, 2016a), and from having a low content of therapeutically active antibodies, particularly against small venom components with low immunogenicity and high toxicity (Laustsen et al, 2017;Leong et al, 2015;Tan et al, 2015Tan et al, , 2016. In recent time, venomics and antivenomics approaches have helped build a greater understanding of the antigenicity of different venom components, which may provide guidance on how to design immunization mixtures that give rise to venom-paraspecific antibody responses in production animals (Calvete et al, 2014(Calvete et al, , 2009Lomonte and Calvete, 2017). Such knowledge creates a foundation for improving the neutralization potential of antivenoms against toxin subfamilies with low immunogenicity, such…”