Cytotoxic Activity of A New Isoform l-Amino Acid Oxidase (Balt-LAAO-II) From Bothrops alternatus (Urutu) Snake Venom in Human Leukemic HL60 Cells

Mauricio Aurelio Gomes Heleno, Alexandre Nowill, João Ernesto de Carvalho, Diego L. Suni-Curasi, Julissa Vilca-Quispe, Emilio Alberto Ponce-Fuentes, Gustavo Alberto Obando-Pereda, Luis Alberto Ponce-Soto

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Resumen

In this study, we describe the isolation of a new isoform, l-Amino acid oxidase (LAAO), referred to as Balt-LAAO-II, from Bothrops alternatus snake venom, which was highly purified using a combination of molecular exclusion (Sephadex G-75) and RP-HPLC chromatographic steps. SDS-PAGE analysis showed that purified Balt-LAAO-II had a molecular weight of ∼ 66 kDa. The N-terminal amino acid sequence and internal peptide sequences showed close structural homology to those of other snake venom l-Amino acid oxidases. This enzyme induces in vitro cytotoxicity in cultured human leukemic HL60 cells. Cells were grown in RPMI medium and incubated with the isoform Balt-LAAO-II (1, 10, and 100 μ g/mL) for up to 72 h. All three concentrations of venom markedly decreased cell viability from 6 h onwards based on staining with propidium iodide, the reduction of 3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and the uptake of neutral red. Flow cytometry showed that all isoforms of Balt-LAAO-II and whole venom concentrations induced apoptosis after 2–6 h of incubation. Morphological analysis of cells incubated with the isoform Balt-LAAO-II and whole venom showed cell rounding and lysis, which increased with venom concentration and duration of incubation. These results show that the isoform Balt-LAAO-II from venom Bothrops alternatus is cytotoxic to cultured HL60 cells, suggesting that this damage may involve the apoptotic and oxidative stress pathways.

Idioma originalInglés
Número de artículo1
PublicaciónInternational Journal of Peptide Research and Therapeutics
Volumen30
N.º1
DOI
EstadoPublicada - ene. 2024

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