Contents lists available at ScienceDirect Journal of Proteomics journal homepage: www.elsevier.com/locate/jprot Global proteomic and functional analysis of Crotalus durissus collilineatus individual venom variation and its impact on envenoming Isadora Sousa de Oliveira a , Iara Aimê Cardoso a , Karla de Castro Figueiredo Bordon a , Sante Emmanuel Imai Carone b , Johara Boldrini-França a , Manuela Berto Pucca c , Karina Furlani Zoccal b , Lúcia Helena Faccioli b , Suely Vilela Sampaio b , José César Rosa d , Eliane Candiani Arantes a, ⁎ a Department of Physics and Chemistry, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil b Department of Clinical Analysis, Toxicology and Food Science, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil c Health Sciences Center, School of Medicine, Federal University of Roraima, Boa Vista, RR, Brazil d 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 ARTICLE INFO Keywords: Snake venoms Crotalus durissus collilineatus Venomic Hyaluronidase activity Myotoxicity 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 glu- tathione 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 varia- tions 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 in- dividual 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. 1. Introduction The Crotalus genus belongs to Viperidae family and is represented in Brazil by the unique species Crotalus durissus, which is subdivided into seven subspecies: C. d. dryinas (Amapá), C. d. terrificus (south region – oriental and meridional, dry and high areas), C. d. cascavella (northeast – caatinga regions), C. d. trigonicus (Roraima), C. d. ruruima (north re- gion), C. d. marajoensis (Marajó Island) and C. d. collilineatus (midwest region, north of Minas Gerais and São Paulo) [1–3]. Snake venoms are composed by proteins [4], which can be divided in enzymatic functions (serine protease, phospholipases and others) and non-enzymatic functions components (growth factors, disintegrins, among others) [5]. In addition, these proteins are responsible for most of signs and symptoms of crotalic envenoming, which affects almost 2,500 victims per year in Brazil [6]. Crotalic envenoming is characterized by an outstanding neurotoxic https://doi.org/10.1016/j.jprot.2018.02.020 Received 27 October 2017; Received in revised form 29 January 2018; Accepted 10 February 2018 ⁎ Corresponding author at: University of São Paulo, School of Pharmaceutical Sciences of Ribeirão Preto, Department of Physics and Chemistry, Av. do Café s/n°, Monte Alegre, 14040- 903 Ribeirão Preto, SP, Brazil. E-mail address: ecabraga@fcfrp.usp.br (E.C. Arantes). Journal of Proteomics xxx (xxxx) xxx–xxx 1874-3919/ © 2018 Elsevier B.V. All rights reserved. Please cite this article as: De Oliveira, I.S., Journal of Proteomics (2018), https://doi.org/10.1016/j.jprot.2018.02.020