Unveiling the Protein Components of the Secretory-Venom Gland and Venom of the Scorpion Centruroides possanii (Buthidae) through Omic Technologies

Abstract

Centruroides possanii is a recently discovered species of “striped scorpion” found in Mexico. Certain species of Centruroides are known to be toxic to mammals, leading to numerous cases of human intoxications in the country. Venom components are thought to possess therapeutic potential and/or biotechnological applications. Hence, obtaining and analyzing the secretory gland transcriptome and venom proteome of C. possanii is relevant, and that is what is described in this communication. Since this is a newly described species, first, its LD50 to mice was determined and estimated to be 659 ng/g mouse weight. Using RNA extracted from this species and preparing their corresponding cDNA fragments, a transcriptome analysis was obtained on a Genome Analyzer (Illumina) using the 76-base pair-end sequencing protocol. Via high-throughput sequencing, 19,158,736 reads were obtained and ensembled in 835,204 sequences. Of them, 28,399 transcripts were annotated with Pfam. A total of 244 complete transcripts were identified in the transcriptome of C. possanii. Of these, 109 sequences showed identity to toxins that act on ion channels, 47 enzymes, 17 protease inhibitors (PINs), 11 defense peptides (HDPs), and 60 in other components. In addition, a sample of the soluble venom obtained from this scorpion was analyzed using an Orbitrap Velos apparatus, which allowed for identification by liquid chromatography followed by mass spectrometry (LC-MS/MS) of 70 peptides and proteins: 23 toxins, 27 enzymes, 6 PINs, 3 HDPs, and 11 other components. Until now, this work has the highest number of scorpion venom components identified through omics technologies. The main novel findings described here were analyzed in comparison with the known data from the literature, and this process permitted some new insights in this field.

Whole transcriptome sequencing reveals the activity of the PLA2 family members in Androctonus crassicauda (Scorpionida: Buthidae) venom gland

“…Finding several homologs of PLA2 with sequence similarity to platelet-activating factor acetylhydrolases (PAFA), cytosolic or calciumdependent PLA2s (cPLA2), secreted PLA2s (sPLA2), and the calcium-independent PLA2s (iPLA2) from venom glands of A. crassicauda is one of the achievements of this study. The findings of this study are in line with previous research that revealed several coding transcripts with a sequence identity of PLA2 from the venom glands of scorpions Centruroides possanii, 36 Liocheles australasiae, Mesobuthus martensii, and Scorpio maurus palmatus using high-throughput sequencing and proteomic analysis. 37 In these studies, phospholipases were the most abundant venom gland enzymes.…”

Characterization of Sodium Channel Peptides Obtained from the Venom of the Scorpion Centruroides bonito

“…The sequences recovered by BLASTP, and phylogenetic analysis, showed that these toxins belong to the subfamily beta of scorpion Na + toxins (β–NaScTx). The amino acid sequence of Cbo1 toxin is identical to antimammalian amidated toxins Cll2b (UniProt accession P59899) from Centruroides limpidus [ 22 ], Cii1 (UniProt accession P59897) from Centruroides infamatus [ 23 ], and CpoNaTBet09 from Centruroides possanii [ 24 ]. The toxins closest to Cbo1 in the phylogenetic tree and with the highest percentage identity are the antimammalian toxins Co1 (UniProt accession C0HLF2) from Centruroides ornatus [ 25 ] and Cb1 (UniProt accession C0HLR3) from Centruroides baergi [ 26 ], with which Cbo1 shares 98% and 97% amino acid identity, respectively ( Figure 3 A,B).…”

“…Sequences recovered from the NCBI are indicated with their accession numbers, followed by toxin name and the species name. CpoNaTBet09 was retrieved from the original publication [ 24 ]. Three α–NaScTx (BTN, Aah3, and Os3) were used as outgroups and to root the tree.…”