Peptidomic and transcriptomic profiling of four distinct spider venoms

Abstract

Venom based research is exploited to find novel candidates for the development of innovative pharmacological tools, drug candidates and new ingredients for cosmetic and agrochemical industries. Moreover, venomics, as a well-established approach in systems biology, helps to elucidate the genetic mechanisms of the production of such a great molecular biodiversity. Today the advances made in the proteomics, transcriptomics and bioinformatics fields, favor venomics, allowing the in depth study of complex matrices and the elucidation even of minor compounds present in minute biological samples. The present study illustrates a rapid and efficient method developed for the elucidation of venom composition based on NextGen mRNA sequencing of venom glands and LC-MS/MS venom proteome profiling. The analysis of the comprehensive data obtained was focused on cysteine rich peptide toxins from four spider species originating from phylogenetically distant families for comparison purposes. The studied species were Heteropoda davidbowie (Sparassidae), Poecilotheria formosa (Theraphosidae), Viridasius fasciatus (Viridasiidae) and Latrodectus mactans (Theridiidae). This led to a high resolution profiling of 284 characterized cysteine rich peptides, 111 of which belong to the Inhibitor Cysteine Knot (ICK) structural motif. The analysis of H. davidbowie venom revealed a high richness in term of venom diversity: 95 peptide sequences were identified; out of these, 32 peptides presented the ICK structural motif and could be classified in six distinct families. The profiling of P. formosa venom highlighted the presence of 126 peptide sequences, with 52 ICK toxins belonging to three structural distinct families. V. fasciatus venom was shown to contain 49 peptide sequences, out of which 22 presented the ICK structural motif and were attributed to five families. The venom of L. mactans, until now studied for its large neurotoxins (Latrotoxins), revealed the presence of 14 cysteine rich peptides, out of which five were ICK toxins belonging to the CSTX superfamily. This in depth profiling of distinct ICK peptide families identified across the four spider species highlighted the high conservation of these neurotoxins among spider families.

Addition of K22 Converts Spider Venom Peptide Pme2a from an Activator to an Inhibitor of NaV1.7

“…Here, we describe the isolation and Na V activity of two novel spider venom-derived peptides, which are the first peptides to be isolated and functionally characterised from the venom of the species P. metallica, as well as the entire Poecilotheria genus [21]. A transcriptome and proteome from the venom gland of the related species P. formosa identified two related, but not identical peptide sequences (Pf29 and Pf32) [22], indicating that the entire Poecilotheria genus is likely to be a rich source of novel Na V modulators ( Figure 2). Indeed, crude venom from several species of the Poecilotheria genus, including P. metallica, has previously been shown to modulate Na V 1.7 [23], consistent with activity of Pme1a and Pme2a at both Na V 1.7 and Na V 1.8 described here.…”

Molecular basis and mechanism underlying the insecticidal activity of venoms and toxins from Latrodectus spiders

“…In a transcriptomic analysis of L. mactans venom, five ICK toxins were discovered. The ICK toxins are believed to ubiquitously present in venomous animals particularly spider venoms, having an essential insecticidal role due to their potent effects on ion channels . When toxins identified from L. hesperus venom were compared with those of L. tredecimguttatus venom, there were many similarities, and few differences.…”

Aspartic Acid Isomerization Characterized by High Definition Mass Spectrometry Significantly Alters the Bioactivity of a Novel Toxin from Poecilotheria

“…For the purpose of this study, we employed a comprehensive high definition mass spectrometry (HDMS E ) approach to investigate the venom from ten species of an arboreal tarantula commonly known as the "tiger" or "ornamental" spider (Genus Poecilotheria): P. striata, P. regalis, P. rufilata, P. formosa, P. metallica, P. miranda, P. tigrinawesseli, P. fasciata, P. ornata, P. smithi, and P. vittata. While there have been studies of whole venom from this genus of aggressive tarantula [46][47][48][49], our laboratory is the first to characterize a complete primary structure of a poecilotheriatoxin as well as evaluate the impact isomerization has on a toxin's physiological effect(s). Using a combination of digestion strategies, fragmentation techniques, and tandem high-resolution ion mobility spectrometry (IMS-interchangeable in this manuscript as high definition mass spectrometry HDMS E ), potential candidates for one non-enzymatic PTM was identified and confirmed, i.e., isomerization of Asp.…”