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

Abstract

Buthus occitanus (B.occitanus) is one of the most dangerous scorpions in the world. Despite the involvement of B.occitanus scorpion in severe cases of envenomation in Morocco, no study has focused yet on the proteomic composition of the Moroccan B.occitanus scorpion venom. Mass spectrometry-based proteomic techniques are commonly used in the study of scorpion venoms. The implementation of top-down and bottom-up approaches for proteomic analyses facilitates screening by allowing a global view of the structural aspects of such complex matrices. Here, we provide a partial overview of the venom of B.occitanus, in order to explore the diversity of its toxins and hereafter understand their effects. To this end, a combination of top-down and bottomup approaches was applied using nano-high liquid chromatography coupled to nano-electrospray tandem mass spectrometry (nano-LC-ESI-MS/MS). TheLC-MS results showed that B.occitanus venom contains around 200 molecular masses ranging from 1868 to 16,720 Da, the most representative of which are those between 5000 and 8000 Da. Interestingly, combined top-down and bottom-up LC-MS/MS results allowed the identification of several toxins, which were mainly those acting on ion channels, including those targeting sodium (NaScTxs), potassium (KScTxs), chloride (ClScTxs) and calcium channels (CaScTx), as well as antimicrobial peptides (AMPs), amphipathic peptides, myotropic neuropeptides and hypothetical secreted proteins. This study reveals the molecular diversity of B.occitanus scorpion venom and identifies components that may have useful pharmacological activities.

Induced pathophysiological alterations by the venoms of the most dangerous Moroccan scorpions Androctonus mauretanicus and Buthus occitanus: A comparative pathophysiological and toxic-symptoms study

“…As for their composition, scorpion venoms are complex matrices, consisting of mucopolysaccharides, hyaluronidases, phospholipases, serotonin and histamine, but mainly of neurotoxins and small peptides 8 (<10 kDa) that target the ionic channels of excitable cells; especially sodium, potassium, calcium, and chloride. 9,10,11 These neurotoxins are the most abundant components in the venoms of scorpions from the Buthidae family and may represent more than 73% of the venom composition. 12 During a scorpion sting, a very small amount of venom is inoculated causing multiple symptom that can be digestive, vegetative, neurological, cardiovascular, and/or respiratory signs 13,14 and can eventually lead to death.…”

Toxinology in the proteomics era: a review on arachnid venom proteomics

“…Although we focused on studies describing the New-World venomics, it is important to highlight that a number of venomic studies from Old-World scorpions have also been published [91,[95][96][97][98][99][100][101].…”

Anti-Cancer Effect of Moroccan Cobra Naja haje Venom and Its Fractions against Hepatocellular Carcinoma in 3D Cell Culture

“…The enzymatic reaction was quenched by adding 5 µL of formic acid (FA) 5%. Then, the digested fraction was dried and resuspended in 0.1% (v:v) FA and 5% (v:v) acetonitrile (ACN) [44].…”

“…The total cycle time was two seconds. In each cycle, five of the most abundant precursor ions were subjected to MS/MS fragmentation; the ions with single charge were excluded, while the collision energy was automatically adjusted according to m/z [44].…”

“…Specific parameters were set as follows: to carry out the research, mass tolerance of parents and fragments set at 50 ppm and 0.3 Da, respectively; enzymatic specificity, trypsin; three as the maximum missed cleavages for trypsin; variable modification, oxidation (M), carbamidomethylation, pyro-glu of Q and E, dehydration and amidation; no fixed modification was taken into account; instrument, ESI-Q-TOF; taxonomy, all; database, UniProtKB/Swiss-Prot. The characterized proteins/peptides have been classified into different families on the basis of their function according to the UniProt database (https://www.uniprot.org, accessed on 23 February 2019) [44].…”