Pharmacological Investigation of CC-LAAO, an L-Amino Acid Oxidase from Cerastes cerastes Snake Venom

Snake venom proteins, which are responsible for deadly snakebite envenomation, induce severe injuries including neurotoxicity, myotoxicity, cardiotoxicity, hemorrhage, and the disruption of blood homeostasis. Yet, many snake-venom proteins have been developed as potential drugs for treating human diseases due to their pharmacological effects. In this study, we evaluated the use of, an L-amino acid oxidase isolated from Cerastes cerastes snake venom CC-LAAO, as a potential anti-glioblastoma drug, by investigating its in vivo and in vitro pharmacological effects. Our results showed that acute exposure to CC-LAAO at 1 and 2.5 µg/mL does not induce significant toxicity on vital organs, as indicated by the murine blood parameters including aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH) activities, and creatinine levels. The histopathological examination demonstrated that only at high concentrations did CC-LAAO induce inflammation and necrosis in several organs of the test subjects. Interestingly, when tested on human glioblastoma U87 cells, CC-LAAO induced a dose-dependent apoptotic effect through the H2O2 generated during the enzymatic reaction. Taken altogether, our data indicated that low concentration of CC-LAAO may be safe and may have potential in the development of anti-glioblastoma agents.

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

10.1101/2020.01.15.907758 ◽

2020 ◽

Author(s):

Maurício Aurelio Gomes Heleno ◽

João Ernesto de Carvalho ◽

Alexandre Nowill ◽

Luis Alberto Ponce-Soto

Keyword(s):

Amino Acid ◽

Snake Venom ◽

Neutral Red ◽

Acid Oxidase ◽

Amino Acid Oxidase ◽

Hl60 Cells ◽

Bothrops Alternatus ◽

Diphenyl Tetrazolium Bromide ◽

Induced Apoptosis

AbstractIn this work 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 chromatographics steps. When analyzed by SDS-PAGE, the purified Balt-LAAO-II presented a molecular weight of ∼66 kDa. The N-terminal amino acid sequence and internal peptide sequences showed close structural homology to other snake venom L-amino acid oxidases.This enzyme induces in vitro cytotoxicity on cultured human leukemic HL60 cells. Cells were grown in RPMI medium and were incubated with isoform Balt-LAAO-II (1, 10 and 100 μg/mL) for up to 72 h. All three concentrations of venom markedly decreased the cell viability from 6 h onwards based on the 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 isoform Balt-LAAO-II and whole venom concentrations induced apoptosis after 2-6 h of incubation. Morphological analysis of cells incubated with isoform Balt-LAAO-II and whole venom showed cell rounding and lysis that increased with the venom concentration and duration of incubation. These results show that isoform Balt-LAAO-II from venom Bothrops alternatus is cytotoxic to cultured HL60 cells and suggest that this damage may involve apoptotic and oxidative stress pathways.

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Bothrops pirajai snake venom L-amino acid oxidase: in vitro effects on infection of Toxoplasma gondii in human foreskin fibroblasts

Revista Brasileira de Farmacognosia ◽

10.1590/s0102-695x2011005000108 ◽

2011 ◽

Vol 21 (3) ◽

pp. 477-485 ◽

Cited By ~ 3

Author(s):

Luiz F. M. Izidoro ◽

Lívia M. Alves ◽

Veridiana M. Rodrigues ◽

Deise A. O. Silva ◽

José R. Mineo

Keyword(s):

Amino Acid ◽

Toxoplasma Gondii ◽

Snake Venom ◽

Acid Oxidase ◽

Amino Acid Oxidase ◽

Human Foreskin Fibroblasts ◽

In Vitro Effects ◽

Bothrops Pirajai

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Impairment of Platelet Aggregation by Echis colorata Venom Mediated by L-Amino Acid Oxidase or H2O2

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657280 ◽

1982 ◽

Vol 48 (03) ◽

pp. 277-282 ◽

Cited By ~ 22

Author(s):

I Nathan ◽

A Dvilansky ◽

T Yirmiyahu ◽

M Aharon ◽

A Livne

Keyword(s):

Hydrogen Peroxide ◽

Amino Acid ◽

Platelet Aggregation ◽

Snake Venom ◽

Oxidase Activity ◽

Enzyme Preparation ◽

Acid Oxidase ◽

Crude Venom ◽

Amino Acid Oxidase ◽

Hemostatic Disorders

SummaryEchis colorata bites cause impairment of platelet aggregation and hemostatic disorders. The mechanism by which the snake venom inhibits platelet aggregation was studied. Upon fractionation, aggregation impairment activity and L-amino acid oxidase activity were similarly separated from the crude venom, unlike other venom enzymes. Preparations of L-amino acid oxidase from E.colorata and from Crotalus adamanteus replaced effectively the crude E.colorata venom in impairment of platelet aggregation. Furthermore, different treatments known to inhibit L-amino acid oxidase reduced in parallel the oxidase activity and the impairment potency of both the venom and the enzyme preparation. H2O2 mimicked characteristically the impairment effects of L-amino acid oxidase and the venom. Catalase completely abolished the impairment effects of the enzyme and the venom. It is concluded that hydrogen peroxide formed by the venom L-amino acid oxidase plays a role in affecting platelet aggregation and thus could contribute to the extended bleeding typical to persons bitten by E.colorata.

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A new l -amino acid oxidase from Bothrops jararacussu snake venom: Isolation, partial characterization, and assessment of pro-apoptotic and antiprotozoal activities

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2017.05.025 ◽

2017 ◽

Vol 103 ◽

pp. 25-35 ◽

Cited By ~ 19

Author(s):

Sante E.I. Carone ◽

Tássia R. Costa ◽

Sandra M. Burin ◽

Adélia C.O. Cintra ◽

Karina F. Zoccal ◽

...

Keyword(s):

Amino Acid ◽

Snake Venom ◽

Partial Characterization ◽

Acid Oxidase ◽

Amino Acid Oxidase ◽

Bothrops Jararacussu

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The anti-ovarian carcinoma activity of L-amino Acid Oxidase from Crotalus adamanteus venom in vivo and in vitro

10.21203/rs.3.rs-1131706/v1 ◽

2021 ◽

Author(s):

Li Bo ◽

Yan Xiong ◽

Qiyi He ◽

Xiaodong Yu ◽

Bo Li ◽

...

Keyword(s):

Ovarian Cancer ◽

Amino Acid ◽

Cancer Cells ◽

Morphological Changes ◽

Complex Mixture ◽

Ovarian Cancer Cells ◽

Acid Oxidase ◽

Amino Acid Oxidase ◽

Apoptosis Pathway

Abstract The anti-tumor potential of animal toxins has fully attracted the attention of researchers. Snake venoms is a complex mixture of different components and has revealed high toxicity on normal and tumoral tissues or cells. The snake venom L-Amino-acid oxidase (svLAAO) has grown up to be a critical research target in molecular biology sciences and medicine sciences since widespread presence and various biological roles, including antitumor application. We found that Crotalus adamanteus (C. adamanteus) venom LAAO significantly decreased the viability of ovarian cancer cells and caused morphological changes preceded cell death. Cell experiments confirmed that C. adamanteus venom LAAO caused alterations of intrinsic or extrinsic apoptosis pathway-related genes in ovarian cancer cells. Animal experiments and histological analysis also proved that C. adamanteus venom LAAO could effectively inhibit the damage of ovarian cancer to tissues. The major apoptosis induction of C. adamanteus venom LAAO on ovarian cancer cells can be blocked by catalase, suggesting that the cytotoxicity of C. adamanteus venom LAAO on ovarian cancer cells was mainly mediated by H2O2. Our preliminary results revealed that C. adamanteus venom LAAO may induce apoptosis of ovarian cancer cells through the death receptor pathway and mitochondrial pathway. It is inferred that C. adamanteus venom LAAO will be some advantages in New Drug Research and Development of antitumor drugs in the future. Nevertheless, extra studies on the pharmacological actions and molecular mechanism of svLAAO in anti-cancer are necessary in order to better promote its application.

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