Two Novel Peptides from Buthotus saulcyi Scorpion Venom: Proteomic Analysis and Approaches

Background and Objectives: Venomous scorpions play a crucial role in medicine and public health. Buthotus saulcyi scorpion is known as one of the most populous species in East Asia and Iran, while its venom proteome has still not been fully determined. background: In the present study, we report for the first time a proteomic profile of Buthotus saulcyi scorpion venom with the aim of looking ahead and determining the structural and functional characteristics of these compounds for use as medical tools. Aims: In the current research, the proteomic profile of Buthotus saulcyi scorpion venom to determine the structural and functional characteristics of its compounds used for treatment will be examined for the first time. objective: In the present study, we report for the first time a proteomic profile of Buthotus saulcyi scorpion venom with the aim of looking ahead and determining the structural and functional characteristics of these compounds for use as medical tools. Method:: 2D-PAGE, HPLC, SDS-PAGE, sequencing, and MALDI-TOF MS techniques were used to investigate the properties of these peptides. Result: The 2D-PAGE analysis of crude toxin from B. saulcyi revealed a minimum of 96 protein spots, with isoelectric points ranging from 4 to 9 and molecular weights spanning from 3.6 to 205 kDa. Following this, HPLC was used to isolate 14 fractions of crude toxins, and the protein content of these fractions was measured. SDS-PAGE analysis identified 7 protein bands within the B. saulcyi crude toxin fractions, with molecular weights ranging from 13 to 217 kDa. Further examination of fraction 7 through amino acid sequencing resulted in the identification of two protein bands labeled peptide 3 and peptide 4. Ultimately, these protein bands were extracted, and their molecular mass and amino acid sequences were analyzed using MALDI-TOF MS. result: 2D-PAGE data from the B. saulcyi crude toxin showed at least 96 protein spots in isoelectric point between 4 and 9 and a molecular weight between 3.6 and 205 kDa. Then 14 fractions of crude toxins were isolated using HPLC and their protein content was estimated. SDS-PAGE analysis of fractions of B. saulcyi crude toxin showed 7 protein bands with a molecular weight between 13 and 217 kDa. Subsequently, the amino acid sequencing of Fraction 7 led us to two protein bands designated as Peptide 3 and Peptide 4 peptides. Finally, these protein bands were removed, and molecular mass and amino acid sequence analysis was performed using MALDI-TOF MS. Conclusion:: According to our results, the alignment of P3 and P4 protein sequences revealed the highest similarity to chrysophsin 2 and pheromone-bound protein 2, respectively.