Global proteomic and functional analysis of Crotalus durissus collilineatus individual venom variation and its impact on envenoming

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de Oliveira, Isadora Sousa ; Cardoso, Iara Aime ; Figueiredo Bordon, Karla de Castro ; Imai Carone, Sante Emmanuel ; Boldrini-Franca, Johara ; Pucca, Manuela Berto ; Zoccal, Karma Furlani ; Faccioli, Lucia Helena ; Sampaio, Suely Vilela ; Rosa, Jose Cesar ; Arantes, Eliane Candiani

• Document type: Journal article
• Source: JOURNAL OF PROTEOMICS; v. 191, p. 13-pg., 2019-01-16.

Abstract

Individual variations studies are important to understand the snakebite envenoming and to improve the anti-venom production and its effectiveness. In this way, the objective of this study was a comparative analysis of intraspecific variation in the venom composition of 22 Crotalus durissus collilineatus specimens through proteomic techniques. Venoms were fractionated by RP-FPLC, and analyzed by SDS-PAGE and mass spectrometry. Although similar, chromatographic and electrophoretic profiles showed significant qualitative and quantitative differences. Some venom components were identified for the very first time in C. d collilineatus, such as glutathione peroxidase, nerve growth factor, 5'-nucleotidase, angiotensin-converting enzyme, carboxypeptidase, phosphodiesterase, glutaminyl cyclase and phospholipase B. Regarding hyaluronidase activity, 2 venoms did not present detectable enzyme activity in the tested amounts. Additionally, in vivo crotalic envenoming in mice showed that venoms from different specimens resulted in diversified changes of biochemical and immunological parameters, such as creatine kinase and interleukin 6. This study demonstrated significant intraspecific variations in the venom of C. d. collilineatus, which may impact the production and effectiveness of the antivenom therapy. Biological significance: This study performed the proteomic and functional analyzes of 22 C. d. collilineatus individual venoms and verified the occurrence of quali and quantitative variations among them. The venoms evaluated caused envenomings with different changes in biochemical and immunological parameters. These results confirm the need to use a pool of venoms with the greatest possible variability in the preparation of antivenoms, in order to improve their effectiveness. In addition, this study was able to identify for the first time 8 different proteins in this subspecies venom, increasing knowledge about its composition and showing that it is a source of these proteins with possible biotechnological applications. (AU)

• FAPESP's process: 14/16182-3 - New functional perspectives of a serine protease from Crotalus durissus collilineatus: activity on voltage-gated ion channels
• Grantee: Johara Boldrini França Stringari
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis
• Grantee: Suely Vilela
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 14/03332-7 - Study of the relationship between inflammasome activation and lipid mediators production with pulmonary inflammation induced by scorpion venom with and without hyaluronidase
• Grantee: Karina Furlani Zoccal
• Support Opportunities: Scholarships in Brazil - Post-Doctoral