Diversity of Phospholipases A from Snake Venom: Adaptive Advantages for Snakes Compromising Treatments for Snakebite Patients

Overview

Journal Toxins (Basel)

Publisher MDPI

Specialty Toxicology

Date 2022 Aug 25

PMID 36006204

Authors

Leijiane F Sousa

Amanda P Freitas

Bruna L Cardoso

Tiago H M Del-Rei

Vanessa A Mendes

Daniele P Orefice

Marisa M T Rocha

Benedito C Prezoto

Ana M Moura-da-Silva

Affiliations

Soon will be listed here.

Abstract

The evolution of snake venoms resulted in multigene toxin families that code for structurally similar isoforms eventually harboring distinct functions. PLAs are dominant toxins in viper venoms, and little is known about the impact of their diversity on human envenomings and neutralization by antivenoms. Here, we show the isolation of three distinct PLAs from venom. FA1 is a Lys-49 homologue, and FA3 and FA4 are catalytic Asp-49 PLAs. FA1 and FA3 are basic myotoxic proteins, while FA4 is an acid non-myotoxic PLA. FA3 was the most potent toxin, inducing higher levels of edema, inflammatory nociception, indirect hemolysis, and anticoagulant activity on human, rat, and chicken plasmas. FA4 presented lower anticoagulant activity, and FA1 had only a slight effect on human and rat plasmas. PLAs presented differential reactivities with antivenoms, with an emphasis on FA3, which was not recognized or neutralized by the antivenoms used in this study. Our findings reveal the functional and antigenic diversity among PLAs from venom, highlighting the importance of assessing venom variability for understanding human envenomations and treatment with antivenoms, particularly evident here as the antivenom fails to recognize FA3, the most active multifunctional toxin described.

Citing Articles

Qualitative Profiling of Venom Toxins in the Venoms of Several Species Using High-Throughput Venomics and Coagulation Bioassaying.

Weekers D, Alonso L, Verstegen A, Slagboom J, Kool J Toxins (Basel). 2024; 16(7).

PMID: 39057940 PMC: 11280908. DOI: 10.3390/toxins16070300.

References

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

Kini R, Rao V, Joseph J . Procoagulant proteins from snake venoms. Haemostasis. 2002; 31(3-6):218-24. DOI: 10.1159/000048066. View

Kimura L, Prezotto-Neto J, Antoniazzi M, Jared S, Santoro M, Barbaro K . Characterization of inflammatory response induced by Potamotrygon motoro stingray venom in mice. Exp Biol Med (Maywood). 2014; 239(5):601-9. DOI: 10.1177/1535370214523704. View

Wen F, Monteiro W, Moura da Silva A, Tambourgi D, da Silva I, Sampaio V . Snakebites and scorpion stings in the Brazilian Amazon: identifying research priorities for a largely neglected problem. PLoS Negl Trop Dis. 2015; 9(5):e0003701. PMC: 4440781. DOI: 10.1371/journal.pntd.0003701. View

Kini R, Chan Y . Accelerated evolution and molecular surface of venom phospholipase A2 enzymes. J Mol Evol. 1999; 48(2):125-32. DOI: 10.1007/pl00006450. View

Sousa L, Nicolau C, Peixoto P, Bernardoni J, Oliveira S, Portes-Junior J . Comparison of phylogeny, venom composition and neutralization by antivenom in diverse species of bothrops complex. PLoS Negl Trop Dis. 2013; 7(9):e2442. PMC: 3772048. DOI: 10.1371/journal.pntd.0002442. View

Hunskaar S, Hole K . The formalin test in mice: dissociation between inflammatory and non-inflammatory pain. Pain. 1987; 30(1):103-114. DOI: 10.1016/0304-3959(87)90088-1. View

Bernardoni J, Sousa L, Wermelinger L, Lopes A, Prezoto B, Serrano S . Functional variability of snake venom metalloproteinases: adaptive advantages in targeting different prey and implications for human envenomation. PLoS One. 2014; 9(10):e109651. PMC: 4196926. DOI: 10.1371/journal.pone.0109651. View

Moura-da-Silva A, Cardoso D, Tanizaki M, Mota I . Neutralization of myotoxic activity of Bothrops venoms by antisera to purified myotoxins and to crude venoms. Toxicon. 1991; 29(12):1471-80. DOI: 10.1016/0041-0101(91)90003-a. View

10.

Gutierrez J, Ownby C . Skeletal muscle degeneration induced by venom phospholipases A2: insights into the mechanisms of local and systemic myotoxicity. Toxicon. 2004; 42(8):915-31. DOI: 10.1016/j.toxicon.2003.11.005. View

11.

Gutierrez J, Calvete J, Habib A, Harrison R, Williams D, Warrell D . Snakebite envenoming. Nat Rev Dis Primers. 2017; 3:17063. DOI: 10.1038/nrdp.2017.63. View

12.

Faure G, Saul F . Structural and Functional Characterization of Anticoagulant, FXa-binding Viperidae Snake Venom Phospholipases A2. Acta Chim Slov. 2013; 58(4):671-7. View

13.

Diaz C, Gutierrez J, Lomonte B, Gene J . The effect of myotoxins isolated from Bothrops snake venoms on multilamellar liposomes: relationship to phospholipase A2, anticoagulant and myotoxic activities. Biochim Biophys Acta. 1991; 1070(2):455-60. DOI: 10.1016/0005-2736(91)90086-n. View

14.

Monteiro W, Contreras-Bernal J, Bisneto P, Sachett J, da Silva I, Lacerda M . , the most important snake involved in human envenomings in the amazon: How venomics contributes to the knowledge of snake biology and clinical toxinology. Toxicon X. 2020; 6:100037. PMC: 7285970. DOI: 10.1016/j.toxcx.2020.100037. View

15.

Landucci E, de Castro R, Toyama M, Giglio J, Marangoni S, Nucci G . Inflammatory oedema induced by the lys-49 phospholipase A(2) homologue piratoxin-i in the rat and rabbit. Effect of polyanions and p-bromophenacyl bromide. Biochem Pharmacol. 2000; 59(10):1289-94. DOI: 10.1016/s0006-2952(00)00248-3. View

16.

Gutierrez J, Rucavado A, Escalante T, Diaz C . Hemorrhage induced by snake venom metalloproteinases: biochemical and biophysical mechanisms involved in microvessel damage. Toxicon. 2005; 45(8):997-1011. DOI: 10.1016/j.toxicon.2005.02.029. View

17.

Amazonas D, Portes-Junior J, Nishiyama-Jr M, Nicolau C, Chalkidis H, Mourao R . Molecular mechanisms underlying intraspecific variation in snake venom. J Proteomics. 2018; 181:60-72. DOI: 10.1016/j.jprot.2018.03.032. View

18.

Zelanis A, Menezes M, Kitano E, Liberato T, Tashima A, Pinto A . Proteomic identification of gender molecular markers in Bothrops jararaca venom. J Proteomics. 2016; 139:26-37. DOI: 10.1016/j.jprot.2016.02.030. View

19.

Nunez V, Arce V, Gutierrez J, Lomonte B . Structural and functional characterization of myotoxin I, a Lys49 phospholipase A2 homologue from the venom of the snake Bothrops atrox. Toxicon. 2004; 44(1):91-101. DOI: 10.1016/j.toxicon.2004.04.013. View

20.

Proleon A, Torrejon D, Urra F, Lazo F, Lopez-Torres C, Fuentes-Retamal S . Functional, immunological characterization, and anticancer activity of BaMtx: A new Lys49- PLA homologue isolated from the venom of Peruvian Bothrops atrox snake (Serpentes: Viperidae). Int J Biol Macromol. 2022; 206:990-1002. DOI: 10.1016/j.ijbiomac.2022.03.111. View