First transcriptome analysis of the venom gland of the scorpion Hottentottta zagrosensis (Scorpion: Buthidae) focusing on venom lipolysis-activating peptides
Introduction: Scorpion venom is a rich source of biologically active peptides and proteins. Transcriptome analysis of venom glands provides detailed insight into the peptide and protein venom components. After conducting transcriptome analysis of various species in previous studies, our research team decided to use the endemic Iranian scorpion to develop biological research focusing on the medical applications of scorpion venom. We focused on Hottentetta zagrosensis, one of the species, and obtained information about its venom proteins. Production of components and antitoxins. To gain insight into the protein composition of this scorpion venom, we performed transcriptome analysis. Methods: Transcriptome analysis of H. zagrosensis venom glands prepared from Khuzestan province was performed by Illumina paired-end sequencing (RNA-Seq). Trinity de novo assembly, CD-Hit-EST clustering, and annotation of identified primary structures using bioinformatics Results: Transcriptome analysis shows the presence of 96.4% of complete arthropod BUSCOs with high quality It was shown that it is a collection of From a total of 45,795,108 paired-end 150 bp trimmed reads, the clustering step generated 101,180 de novo assembled transcripts with an N50 size of 1,149 bp. 96,071 Unigenes and 131,235 transcripts showed significant similarity (E-value 1e-3) to known proteins from UniProt, Swissprot, Animal Toxin Annotation Project, and Pfam databases. Results were verified using InterProScan. These are mainly ion channel inhibitors, metalloproteinases, neurotoxins, protease inhibitors, protease activators, cysteine-rich secreted proteins, phospholipase A enzymes, antimicrobial peptides, growth factors, lipolysis-activating peptides, hyaluronidase, etc. We will respond. Phospholipase D. Our venom gland transcriptomic approach identified several biologically active peptides, including the five LVP1-alpha and LVP1-beta isoforms. We named these HzLVP1_alpha1, HzLVP1_alpha2, HzLVP1_alpha3, HzLVP1_beta1, and HzLVP1_beta and thoroughly characterized them here. <40). Furthermore, all alpha and beta subunit isoforms of LVP1 are thermostable, with HzLVP1_alpha2 (aliphatic index = 71.38) being the most stable. HzLVP1_alpha2 also has the longest half-life, and the three-dimensional structure of all identified proteins is compacted with three disulfide bridges. The extra cysteine ​​residues may allow the protein to form heterodimers or homodimers. The LVP1 subunit of H. zagrosensis can interact with adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), two key enzymes in the control of lipolysis in adipocytes. Pharmacological properties of these identified proteins have been suggested.
