Autor(es): Bittencourt Rodrigues, Caroline Fabri ; Serino-Silva, Caroline ; Morais-Zani, Karen de ; Kavazoi, Victor Koiti ; Nogueira Carvalho, Marcelo Pires ; Grego, Kathleen Fernandes ; Chiarelli, Tassia ; Tashima, Alexandre Keiji ; Toyama, Marcos Hikari [UNESP] ; Tanaka-Azevedo, Anita Mitico
Data: 2020
Identificador Persistente: http://hdl.handle.net/11449/196909
Origem: Oasisbr
Made available in DSpace on 2020-12-10T20:00:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-02-27
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
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.
Univ Sao Paulo, Inst Pesquisas Tecnol, Inst Butantan, Interunidades Biotecnol, Sao Paulo, SP, Brazil
Inst Butantan, Lab Herpetol, Sao Paulo, SP, Brazil
Univ Fed Minas Gerais, Belo Horizonte, MG, Brazil
Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, Brazil
Univ Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP, Brazil
Univ Estadual Paulista, Inst Biociencias Litoral Paulista, Sao Vicente, SP, Brazil
CAPES: 001
FAPESP: 2017/16908-2
FAPESP: 2017/20106-9
FAPESP: 2018/25786-0
