Boa.PLI: Structural and functional characterization of the gamma phospholipase A2 plasma inhibitor from the non-venomous Brazilian snake Boa constrictor

Author(s)

Bittencourt Rodrigues, Caroline Fabri ^{[1, 2]} ; Serino-Silva, Caroline ^{[1, 2]} ; de Morais-Zani, Karen ^{[1, 2]} ; Kavazoi, Victor Koiti ^[1] ; Nogueira Carvalho, Marcelo Pires ^[3] ; Grego, Kathleen Fernandes ^[1] ; Chiarelli, Tassia ^[4] ; Tashima, Alexandre Keiji ^[4] ; Toyama, Marcos Hikari ^[5] ; Tanaka-Azevedo, Anita Mitico ^{[1, 2]}

Affiliation

^[1] Inst Butantan, Lab Herpetol, Sao Paulo, SP - Brazil

^[2] Univ Sao Paulo, Inst Pesquisas Tecnol, Inst Butantan, Interunidades Biotecnol, Sao Paulo, SP - Brazil

^[3] Univ Fed Minas Gerais, Belo Horizonte, MG - Brazil

^[4] Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP - Brazil

^[5] Univ Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP - Brazil

• Document type: Journal article
• Source: PLoS One; v. 15, n. 2 FEB 27 2020.
• Web of Science Citations: 0

Abstract

Plasma in several organisms has components that promote resistance to envenomation by inhibiting specific proteins from snake venoms, such as phospholipases A(2) (PLA(2)s). The major hypothesis for inhibitor's presence would be the protection against self-envenomation in venomous snakes, but the occurrence of inhibitors in non-venomous snakes and other animals has opened new perspectives for this molecule. Thus, this study showed for the first time the structural and functional characterization of the PLA(2) inhibitor from the Boa constrictor serum (Boa.PLI), a non-venomous snake that dwells extensively the Brazilian territory. Therefore, the inhibitor was isolated from B. constrictor serum, with 0.63% of recovery. SDS-PAGE showed a band at similar to 25 kDa under reducing conditions and similar to 20 kDa under non-reducing conditions. Chromatographic analyses showed the presence of oligomers formed by Boa.PLI. Primary structure of Boa.PLI suggested an estimated molecular mass of 22 kDa. When Boa.PLI was incubated with Asp-49 and Lys-49 PLA(2) there was no severe change in its dichroism spectrum, suggesting a non-covalent interaction. The enzymatic assay showed a dose-dependent inhibition, up to 48.2%, when Boa.PLI was incubated with Asp-49 PLA(2), since Lys-49 PLA(2) has a lack of enzymatic activity. The edematogenic and myotoxic effects of PLA(2)s were also inhibited by Boa.PLI. In summary, the present work provides new insights into inhibitors from non-venomous snakes, which possess PLIs in their plasma, although the contact with venom is unlikely. (AU)

• FAPESP's process: 17/20106-9 - Peptidomics of Brazilian snake and spider venoms
• Grantee: Alexandre Keiji Tashima
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/16908-2 - Proteomic characterization and enzymatic and pathophysiological activities of the venom of the snake species that compose the Bothrops neuwiedi group
• Grantee: Karen de Morais Zani
• Support Opportunities: Regular Research Grants
• FAPESP's process: 18/25786-0 - Phospholipase A2 inhibitors (PLIs) present in venomous and non-venomous snake plasmas and evaluation of the neutralizing activity of these inhibitors on the phospholipase A2 (PLA2) activities of snake venoms
• Grantee: Anita Mitico Tanaka-Azevedo
• Support Opportunities: Regular Research Grants