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Adriano M. Morgon, Matheus R. Belisrio, Gabriel O. Meissner, Larissa Vuitika, Dilza Trevisan-Silva, Andrea S. Ribeiro, Luiza H. Gremski, Olga M. Chaim, Waldemiro Gremski, Silvio S. Veiga (Federal University of Parana, Curitiba, Brazil)
The Loxosceles Astacin-Like Proteins (LALPs) are astacin-like metalloproteases present in brown-spider venoms. Generally astacins are involved in processes such as embryogenesis, tissue morphogenesis, and others. However, after spiders envenomation these proteins cause deleterious effects. Three isoforms of LALP were cloned from Loxosceles intermedia, yet only one was expressed and evaluated according to its biological activity. Bioinformatics tools have been essential for further studies regarding other isoforms of this enzyme. NCBI tools as the PubMed database and the sequence alignment tool BLAST helped provide data about LALPs and other metalloproteases. Still, tools like ProtParam allowed a deeper analysis of characteristics of the new LALP isoform being studied. Softwares such as QuantityOne and ChromasPro helped in cloning, analysis and confirmation of the sequence of the protein. Also, tools like ClustalW made possible the alignment of the sequence obtained from the cloning with the sequence from the NCBI database. Finally, the Expasy Translate Tool allowed the analysis of the correct reading frame after cloning, aiming for the protein expression in prokaryote systems.