In vitro cytotoxicity of L-amino acid oxidase from the venom of Crotalus mitchellii pyrrhus

Toxicon ◽  
2017 ◽  
Vol 139 ◽  
pp. 20-30 ◽  
Author(s):  
Kok Keong Tan ◽  
Siok Ghee Ler ◽  
Jayantha Gunaratne ◽  
Boon Huat Bay ◽  
Gopalakrishnakone Ponnampalam
Keyword(s):  
Amino Acid ◽  
In Vitro Cytotoxicity ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Crotalus Mitchellii

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

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.

scholarly journals Evaluation of human D-amino acid oxidase inhibition by anti-psychotic drugs in vitro

2014 ◽  
Vol 8 (3) ◽  
pp. 149-154 ◽  
Author(s):  
Miho Shishikura ◽  
Hitomi Hakariya ◽  
Sumiko Iwasa ◽  
Takashi Yoshio ◽  
Hideaki Ichiba ◽  
...  
Keyword(s):  
Amino Acid ◽  
Acid Oxidase ◽  
Anti Psychotic ◽  
Amino Acid Oxidase ◽  
Oxidase Inhibition

scholarly journals Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea Mycelium in Culture Conditions

2012 ◽  
Vol 102 (11) ◽  
pp. 1054-1063 ◽  
Author(s):  
Chi-Hua Cheng ◽  
Chia-Ann Yang ◽  
Kou-Cheng Peng
Keyword(s):  
Amino Acid ◽  
Botrytis Cinerea ◽  
Trichoderma Harzianum ◽  
Hyphal Growth ◽  
Extracellular Proteins ◽  
In Vitro Assays ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Trichoderma Spp

Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea.

scholarly journals Snake venomics of Bothrops punctatus, a semi-arboreal pitviper species from Antioquia, Colombia

2013 ◽  
Author(s):  
Maritza Fernández Culma ◽  
Jaime A Pereañez ◽  
Vitelbina Núñez Rangel ◽  
Bruno Lomonte
Keyword(s):  
Amino Acid ◽  
Protein Composition ◽  
Secretory Proteins ◽  
Acid Oxidase ◽  
Crude Venom ◽  
Amino Acid Oxidase ◽  
Analytical Strategy ◽  
Phospholipases A2 ◽  
Snake Venomics

Bothrops punctatus is an endangered, semi-arboreal pitviper species distributed in Panamá, Colombia, and Ecuador, whose venom is poorly characterized. In the present work, the protein composition of this venom was profiled using the 'snake venomics' analytical strategy. Decomplexation of the crude venom by RP-HPLC and SDS-PAGE, followed by tandem mass spectrometry of tryptic digests, showed that it consists of proteins assigned to at least nine snake toxin families. Metalloproteinases are predominant in this secretion (41.5% of the total proteins), followed by C-type lectin/lectin-like proteins (16.7%), bradykinin-potentiating peptides (10.7%), phospholipases A2 (9.3%), serine proteinases (5.4%), disintegrins (3.8%), L-amino acid oxidases (3.1%), vascular endothelial growth factors (1.7%), and cysteine-rich secretory proteins (1.2%). Altogether, 6.6% of the proteins were not identified. In vitro, the venom exhibited proteolytic, phospholipase A2, and L-amino acid oxidase activities, as well as angiotensin-converting enzyme (ACE)-inhibitory activity, in agreement with the obtained proteomic profile. Cytotoxic activity on murine C2C12 myoblasts was negative, suggesting that the majority of venom phospholipases A2 likely belong to the acidic type, which often lack major toxic effects. The protein composition of B. punctatus venom shows a good correlation with toxic activities here and previously reported, and adds further data in support of the wide diversity of strategies that have evolved in snake venoms to subdue prey, as increasingly being revealed by proteomic analyses.

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

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.

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

2011 ◽  
Vol 21 (3) ◽  
pp. 477-485 ◽  
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

scholarly journals Protective Role of d-Amino Acid Oxidase against Staphylococcus aureus Infection

2012 ◽  
Vol 80 (4) ◽  
pp. 1546-1553 ◽  
Author(s):  
Hideaki Nakamura ◽  
Jun Fang ◽  
Hiroshi Maeda
Keyword(s):  
Hydrogen Peroxide ◽  
Staphylococcus Aureus ◽  
Amino Acid ◽  
Bactericidal Activity ◽  
Hypochlorous Acid ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Content Type

ABSTRACTd-Amino acid oxidase (DAO) is a hydrogen peroxide-generating enzyme that uses ad-amino acid as a substrate. We hypothesized that DAO may protect against bacterial infection, because hydrogen peroxide is one of the most important molecules in the antibacterial defense systems in mammals. We show here that DAO suppressed the growth ofStaphylococcus aureusin a manner that depended on the concentration of DAO andd-amino acidin vitro. Addition of catalase abolished the bacteriostatic activity of DAO. Although DAO plusd-Ala showed less bactericidal activity, addition of myeloperoxidase (MPO) greatly enhanced the bactericidal activity of DAO. Furthermore, DAO was able to utilize bacterial lysate, which containsd-Ala derived from peptidoglycan; this could produce hydrogen peroxide with, in the presence of myeloperoxidase, formation of hypochlorous acid. This concerted reaction of DAO and MPO led to the bactericidal action.In vivoexperiments showed that DAO−/−(mutant) mice were more susceptible toS. aureusinfection than were DAO+/+(wild-type) mice. These results suggest that DAO, together with myeloperoxidase, may play an important role in antibacterial systems in mammals.

Identification of Novel d-Amino Acid Oxidase Inhibitors by in Silico Screening and Their Functional Characterization in Vitro

2013 ◽  
Vol 56 (5) ◽  
pp. 1894-1907 ◽  
Author(s):  
Masumi Katane ◽  
Naoko Osaka ◽  
Satsuki Matsuda ◽  
Kazuhiro Maeda ◽  
Tomonori Kawata ◽  
...  
Keyword(s):  
Amino Acid ◽  
In Silico ◽  
Functional Characterization ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
In Silico Screening
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