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

Isadora Sousa de Oliveira; Iara Aimê Cardoso; Karla de Castro Figueiredo Bordon; Sante Emmanuel Imai Carone; Johara Boldrini-França; Manuela Berto Pucca; Karina Furlani Zoccal; Lúcia Helena Faccioli; Suely Vilela Sampaio; José César Rosa; Eliane Candiani Arantes

doi:10.1016/j.jprot.2018.02.020

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

J Proteomics. 2019 Jan 16:191:153-165. doi: 10.1016/j.jprot.2018.02.020. Epub 2018 Feb 17.

Affiliations

¹ Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
² Department of Clinical Analysis, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
³ Health Sciences Center, School of Medicine, Federal University of Roraima, Boa Vista, RR, Brazil.
⁴ Protein Chemistry Center and Department of Molecular and Cell Biology and Pathogenic Bioagents, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
⁵ Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil. Electronic address: ecabraga@fcfrp.usp.br.

PMID: 29462664
DOI: 10.1016/j.jprot.2018.02.020

Abstract

Individual variations studies are important to understand the snakebite envenoming and to improve the antivenom 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.

Keywords: Crotalus durissus collilineatus; Hyaluronidase activity; Myotoxicity; Snake venoms; Venomic.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Biodiversity
Chromatography, Reverse-Phase
Crotalid Venoms / analysis*
Crotalid Venoms / chemistry
Crotalid Venoms / enzymology
Crotalid Venoms / pharmacology
Crotalus*
Electrophoresis, Polyacrylamide Gel
Mass Spectrometry
Mice
Proteomics / methods*
Snake Bites
Species Specificity

Substances

Crotalid Venoms