Transcriptomic and Proteomic Analyses Reveal the Diversity of Venom Components from the Vaejovid Scorpion Serradigitus gertschi

Abstract

To understand the diversity of scorpion venom, RNA from venomous glands from a sawfinger scorpion, Serradigitus gertschi, of the family Vaejovidae, was extracted and used for transcriptomic analysis. A total of 84,835 transcripts were assembled after Illumina sequencing. From those, 119 transcripts were annotated and found to putatively code for peptides or proteins that share sequence similarities with the previously reported venom components of other species. In accordance with sequence similarity, the transcripts were classified as potentially coding for 37 ion channel toxins; 17 host defense peptides; 28 enzymes, including phospholipases, hyaluronidases, metalloproteases, and serine proteases; nine protease inhibitor-like peptides; 10 peptides of the cysteine-rich secretory proteins, antigen 5, and pathogenesis-related 1 protein superfamily; seven La1-like peptides; and 11 sequences classified as “other venom components”. A mass fingerprint performed by mass spectrometry identified 204 components with molecular masses varying from 444.26 Da to 12,432.80 Da, plus several higher molecular weight proteins whose precise masses were not determined. The LC-MS/MS analysis of a tryptic digestion of the soluble venom resulted in the de novo determination of 16,840 peptide sequences, 24 of which matched sequences predicted from the translated transcriptome. The database presented here increases our general knowledge of the biodiversity of venom components from neglected non-buthid scorpions.

Dissecting Toxicity: The Venom Gland Transcriptome and the Venom Proteome of the Highly Venomous Scorpion Centruroides limpidus (Karsch, 1879)

“…Nevertheless, the results here reported are comparable with those generated for other non-buthid scorpions under the exact same experimental protocol. That is the case for the analyses performed with Serradigitus gertschi , Superstitionia donensis , T. atrox , P. schwenkmeyeri , and M. gertschi [7,10,12,24,25]. As Figure 7A illustrates, the venom of C. limpidus is characterized by the highest diversity of toxins, in general.…”

“…A total of 9 µg (males) and 8 µg (females) of RNA was obtained and its quality was assessed with a Bioanalyzer 2100 (Agilent), as reported in other transcriptomic analyses [7,10,12,24,25]. The results indicated that the RNA samples were not degraded, even though a single RNA band corresponding to the mitochondrial 18S was observed, an effect previously reported for other organisms [7,10,12,24,25]. Paired-end cDNA libraries were prepared for males and females separately and sequenced using the Illumina platform (2 × 72 bp reads).…”

“…They are characterized by having a broad spectrum of biological activities, including antimicrobial [55,56,57], insecticidal [58], bradykinin-potentiating [59], antitumoral [60,61], and hemolytic [62], among others. Host defense peptides have been demonstrated to be abundant and highly diverse in the venoms of scorpions belonging to non-Buthidae families [7,24,25], though a previous study with high-throughput sequencing techniques also identified HDPs in the buthid C. hentzi [8]. Host defense peptides are divided into two categories: non-disulfide-bridged-peptides (NDBPs) and cysteine-stabilized β-sheet-rich peptides, which include the defensins and scorpines [54].…”

Proteo-Transcriptomic Analysis Identifies Potential Novel Toxins Secreted by the Predatory, Prey-Piercing Ribbon Worm Amphiporus lactifloreus

“…The third group encodes mucus-related proteins (mucins) that have also been found in amphibian skin secretions [ 29 ]. The fourth group encodes four toxin protein families: (1) IGFBP-like proteins (insulin-like growth factor binding proteins), which are found in the venoms of invertebrates, such as scorpions and remipedes [ 24 , 30 ]; (2) plancitoxins, previously identified in the crown-of-thorns starfish, a venomous echinoderm [ 31 , 32 ]; (3) actitoxins, previously identified in sea anemones [ 33 ]; (4) ShK-like proteins ( Stichodactyla -like proteins) that are also major venom components in sea anemones [ 34 ]. The expression levels of each transcript are presented in relative values as a pie chart ( Figure 2 A) and also in absolute TPM values as a bar chart ( Figure 2 B).…”

The Dual α-Amidation System in Scorpion Venom Glands

“…Previously reported transcriptomic analyses from venom glands of the scorpion species C. limpidus, Paravaejovis schwenkmeyeri, Urodacus yaschenkoi, Thorellius cristimanus (reported as T. atrox), Serradigitus gertschi, S. donensis, and Megacormus gertschi [37,48,49,50,51,52,53] were used to obtain relevant sequence information. Complementary sequence information was obtained from other unpublished transcriptomes for the species Centruroides noxius, C. orizaba, C. ochraceus, C. hirsutipalpus, T. trivittatus, L. abdullahbayrami, Hoffmannihadrurus aztecus, Hadrurus concolorus, Anuroctonus pococki bajae, Chihuahuanus coahuilae and Diplocentrus melici.…”