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

Abstract

Venom glands and soluble venom from the Mexican scorpion Centruroides limpidus (Karsch, 1879) were used for transcriptomic and proteomic analyses, respectively. An RNA-seq was performed by high-throughput sequencing with the Illumina platform. Approximately 80 million reads were obtained and assembled into 198,662 putative transcripts, of which 11,058 were annotated by similarity to sequences from available databases. A total of 192 venom-related sequences were identified, including Na+ and K+ channel-acting toxins, enzymes, host defense peptides, and other venom components. The most diverse transcripts were those potentially coding for ion channel-acting toxins, mainly those active on Na+ channels (NaScTx). Sequences corresponding to β- scorpion toxins active of K+ channels (KScTx) and λ-KScTx are here reported for the first time for a scorpion of the genus Centruroides. Mass fingerprint corroborated that NaScTx are the most abundant components in this venom. Liquid chromatography coupled to mass spectometry (LC-MS/MS) allowed the identification of 46 peptides matching sequences encoded in the transcriptome, confirming their expression in the venom. This study corroborates that, in the venom of toxic buthid scorpions, the more abundant and diverse components are ion channel-acting toxins, mainly NaScTx, while they lack the HDP diversity previously demonstrated for the non-buthid scorpions. The highly abundant and diverse antareases explain the pancreatitis observed after envenomation by this species.

Mass spectrometry‐based top‐down and bottom‐up approaches for proteomic analysis of the Moroccan Buthus occitanus scorpion venom

“…Top‐down data sets showed that the venom of B. occitanus is very complex, counting around 200 MWs ranging from 1868 to 16 720 Da. A similar number of components have been revealed by previous studies [32–34], others showed fewer components, as well as Leiurus abdullahbayrami (45 masses) and Opisthacanthus elatus (106 masses) [35, 36], whereas some other scorpion venoms were more complex, such as the Pandinus cavimanus (390 masses) and Centruroides limpidus (395 masses) [37, 38]. Additionally, the repartition of MWs showed that < 1% were components with molecular masses < 2000 Da, 14% were those from 2000 to 5000 Da, 74% were those between 5000 and 8000 Da, and 10% were those over than 8000 Da, while the repartition of MW from the French B. occitanus scorpion venom showed an abundance of molecules ranging from 2000 to 3000 Da and those less than 2000 Da [39].…”

“…The major representative category of components identified in our venom were neurotoxins, mainly NaScTxs (77%), these neurotoxins are abundant in species from the Buthidae family [38,46,47] and less representative in scorpions from the non-Buthidae family [33,48,49]. Those toxins are the ones responsible for envenomation symptoms [39]; their high content in [17,66], and hypothetical secreted protein which are proteins with unknown activities.…”

“…Centruroides limpidus (395 masses) [37,38]. Additionally, the repartition of molecular weights showed that less than 1% were components with molecular masses <2000 Da, 14% those from 2000 to 5000 Da, 74% those between 5000 to 8000 Da, and 10% were those over than 8000 Da, while the repartition of molecular weights from the French B.occitanus scorpion venom showed an abundance of molecules ranging from 2000 to 3000 Da and those less than 2000 Da [39].…”

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.…”

Arthropod Venom Components and Their Potential Usage

“…To dissect venom cocktails, particularly when limited amounts of crude venom are available from tiny animals, as in the case of most arthropod species, omics technologies have demonstrated to be an essential collection of robust strategies. Indeed, transcriptome and proteome, alone or in combination with functional analysis, has been applied to disclose and resolve the toxin peptide complexity of the venom, as described from the highly venomous Mexican scorpion Centruroides limpidus [4], the predatory giant ant Dinoponera quadriceps [5], and the predatory ant Odontomachus monticola [6]. In a later study published in this special issue, the authors also investigated the components of the O. monticola venom sac, besides the crude venom.…”