l-Amino acid oxidase from Cerastes vipera snake venom: Isolation, characterization and biological effects on bacteria and tumor cell lines

Salama, Walaa H.; Ibrahim, Nihal M.; El-Hakim, Amr E.; Bassuiny, Roqaya I.; Mohamed, Manal M.; Mousa, Fatma M.; Ali, Mamdouh M.

doi:10.1016/j.toxicon.2018.06.064

Cited by 33 publications

(30 citation statements)

References 58 publications

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“…In general, SV-LAAOs are highly sensitive to changes in temperature and pH; therefore, the usual suggestion is that these enzymes should be stored in neutral pH solutions at 4 °C [ 1 ]. Studies indicate that SV-LAAOs may remain active for variable periods of time over a wide temperature range (0 to 50 °C), with exposure to temperatures above 55 °C resulting in a gradual decrease in activity caused by disruptions in hydrophobic interactions and hydrogen bonds between different subunits of the enzyme [ 7 , 9 , 11 , 15 , 23 , 24 ].…”

Section: Resultsmentioning

confidence: 99%

“…L-amino acid oxidases (LAAOs) are flavoenzymes found in different organisms; however, those from snake venoms (SV-LAAOs) are the most well-characterized enzymes of this protein family [ 1 – 3 ]. In recent decades, many studies have been dedicated to exploring the physical-chemical properties, structural characteristics and biological functions of SV-LAAOs from different snake species [ 3 – 7 ]. Such studies indicate that SV-LAAOs from different sources can present significant variances in molecular mass, substrate specificity, stability, enzymatic and biological activity [ 1 , 8 , 9 ].…”

Section: Introductionmentioning

confidence: 99%

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Kinetic investigations and stability studies of two Bothrops L-amino acid oxidases

Costa

Carone

Tucci

et al. 2018

J Venom Anim Toxins Incl Trop Dis

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BackgroundL-amino acid oxidases isolated from snake venoms (SV-LAAOs) are enzymes that have great therapeutic potential and are currently being investigated as tools for developing new strategies to treat various diseases, including cancer and bacterial infections. The main objective of this study was to make a brief evaluation of the enzymatic stability of two Bothrops LAAOs, one isolated from Bothrops jararacussu (BjussuLAAO-II) and the other from Bothrops moojeni (BmooLAAO-I) venoms.Methods and resultsThe enzymatic activity and stability of both LAAOs were evaluated by microplate colorimetric assays, for which BjussuLAAO-II and BmooLAAO-I were incubated with different L-amino acid substrates, in the presence of different ions, and at different pH ranges and temperatures. BjussuLAAO-II and BmooLAAO-I demonstrated higher affinity for hydrophobic amino acids, such as Phe and Leu. The two enzymes showed high enzymatic activity in a wide temperature range, from 25 to 75 °C, and presented optimum pH around 7.0. Additionally, Zn2+, Al3+, Cu2+ and Ni2+ ions negatively modulated the enzymatic activity of both LAAOs. As to stability, BjussuLAAO-II and BmooLAAO-I showed high enzymatic activity for 42 days stored at 4 °C in neutral pH solution. Moreover, the glycan portions of both LAAOs were analyzed by capillary electrophoresis, which revealed that BjussuLAAO-II presented two main glycan portions with relative masses of 7.78 and 8.13 CGU, while BmooLAAO-I showed three portions of 7.58, 7.94 and 8.37 CGU.ConclusionsOur results showed that, when stored properly, BjussuLAAO-II and BmooLAAO-I present enzymatic stability over a long time period, which is very important to allow the use of these enzymes in pharmacological studies of great impact in the medical field.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Kinetic investigations and stability studies of two Bothrops L-amino acid oxidases

Costa

Carone

Tucci

et al. 2018

J Venom Anim Toxins Incl Trop Dis

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“…It should be mentioned that the cytotoxicity against MCF7 and A549 cells were manifested by several snake venom proteins other than phospholipases A 2 . Cytotoxins from cobra venoms (Ebrahim et al, 2015;Attarde & Pandit, 2017), ruviprase from Russell's viper (Thakur et al, 2016), and L-amino acid oxidases (Li et al, 2013;Salama et al, 2018) are found to inhibit the proliferation of MCF7 cells with different potency. The highest activity demonstrated L-amino acid oxidase from king cobra venom with an IC50 value of 0.04 μg/mL in MCF7 after 72-hr treatment (Li et al, 2013).…”

Section: Cytotoxic Activitymentioning

confidence: 99%

“…There are numerous communications on cytotoxic effects of venoms from cobras, vipers and pit-vipers on tumour cells (e.g., Zainal Abidin et al, 2019;Nalbantsoy et al, 2017;Ghazaryan et al, 2015). Several proteins, including cytotoxins (Dubovskii & Utkin, 2015), L-amino acid oxidases (Salama et al, 2018), phospholipases A 2 (Sobrinho et al, 2016), disintegrins (Arruda Macêdo et al, 2015) and others manifesting anti-proliferative activity were isolated from these venoms. These proteins themselves, due to their inherent adverse properties (high molecular mass, high toxicity etc.…”

Section: Introductionmentioning

confidence: 99%

Peer Review #1 of "Phospholipase A2 from krait Bungarus fasciatus venom induces human cancer cell death in vitro (v0.1)"

2019

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Background. Snake venoms are the complex mixtures of different compounds manifesting a wide array of biological activities. The venoms of kraits (genus Bungarus, family Elapidae) induce mainly neurological symptoms; however, these venoms show a cytotoxicity against cancer cells as well. This study was conducted to identify in Bungarus fasciatus venom an active compound(s) exerting cytotoxic effects toward MCF7 human breast cancer cells and A549 human lung cancer cells. Methods. The crude venom of Bungarus fasciatus was separated by gel-filtration on Superdex HR 75 column and reversed phase HPLC on C18 column. The fractions obtained were screened for cytotoxic effect against MCF7, A549, and HK2 cell lines using colorimetric assay with the tetrazolium dye MTT-3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The primary structure of active protein was established by ultra high resolution LC-MS/MS. The molecular mechanism of the isolated protein action on MCF7 cells was elucidated by flow cytometry. Results. MTT cell viability assays of cancer cells incubated with fractions isolated from Bungarus fasciatus venom revealed a protein with molecular mass of about 13 kDa possessing significant cytotoxicity. This protein manifested the dose and time dependent cytotoxicity for MCF7 and A549 cell lines while showed no toxic effect on human normal kidney HK2 cells. In MCF7, flow cytometry analysis revealed a decrease in the proportion of Ki-67 positive cells. As Ki-67 protein is a cellular marker for proliferation, its decline indicates the reduction in the proliferation of MCF7 cells treated with the protein. Flow cytometry analysis of MCF7 cells stained with propidium iodide and Annexin V conjugated with allophycocyanin showed that a probable mechanism of cell death is apoptosis. Mass spectrometric studies showed that the cytotoxic protein was phospholipase A 2. The amino acid sequence of this enzyme earlier was deduced from cloned cDNA and in this work it was isolated from the venom as a protein for the first time. It is also the first krait phospholipase A 2 manifesting the cytotoxicity for cancer cells.

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“…These enzymes have anti-cancer (Sun et al, 2003;Lee et al, 2014;Tássia et al, 2017) and anti-HIV activity (Sant'Ana et al, 2008) and may be used as therapeutic agents in many disease conditions like anti-cancer and anti-HIV drugs (Sakurai et al, 2003;Zhang et al, 2004;Teixeira et al, 2016;Tan et al, 2017;Costa et al, 2017;Salama et al, 2018;Tan et al, 2018) (Sun et al, 2003;Zhang and Wei, 2007;Lee et al, 2014;Costa et al, 2014;Tássia et al, 2017;Costa et al, 2017). Besides snake venom, LAAO has been found in the insects, fungi (Nuutinen and Timonen, 2008;Yang et al, 2009;Žun et al, 2017), green algae (Schriek et al, 2009), bacteria (Arima et al, 2009), plants (Nishizawa et al, 2005) and mammals (Blanchard et al, 1944;Du and Clemetson, 2002;Kasai et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Structure–Function Studies and Mechanism of Action of Snake Venom L-Amino Acid Oxidases

Ullah

2020

Front. Pharmacol.

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Snake venom L-amino acid oxidases (SV-LAAOs) are the least studied venom enzymes. These enzymes catalyze the stereospecific oxidation of an L-amino acid to their corresponding a-keto acid with the liberation of hydrogen peroxide (H 2 O 2) and ammonia (NH 3). They display various pathological and physiological activities including induction of apoptosis, edema, platelet aggregation/inhibition, hemorrhagic, and anticoagulant activities. They also show antibacterial, antiviral and leishmanicidal activity and have been used as therapeutic agents in some disease conditions like cancer and anti-HIV drugs. Although the crystal structures of six SV-LAAOs are present in the Protein Data Bank (PDB), there is no single article that describes all of them in particular. To better understand their structural properties and correlate it with their function, the current work describes structure characterization, structure-based mechanism of catalysis, inhibition and substrate specificity of SV-LAAOs. Sequence analysis indicates a high sequence identity (>84%) among SV-LAAOs, comparatively lower sequence identity with Pig kidney D-amino acid oxidase (<50%) and very low sequence identity (<24%) with bacterial LAAOs, Fugal (L-lysine oxidase), and Zea mays Polyamine oxidase (PAAO). The three-dimensional structure of these enzymes are composed of three-domains, a FAD-binding domain, a substrate-binding domain and a helical domain. The sequence and structural analysis indicate that the amino acid residues in the loops vary in length and composition due to which the surface charge distribution also varies that may impart variable substrate specificity to these enzymes. The active site cavity volume and its average depth also vary in these enzymes. The inhibition of these enzymes by synthetic inhibitors will lead to the production of more potent antivenoms against snakebite envenomation.

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l-Amino acid oxidase from Cerastes vipera snake venom: Isolation, characterization and biological effects on bacteria and tumor cell lines

Cited by 33 publications

References 58 publications

Kinetic investigations and stability studies of two Bothrops L-amino acid oxidases

Kinetic investigations and stability studies of two Bothrops L-amino acid oxidases

Peer Review #1 of "Phospholipase A2 from krait Bungarus fasciatus venom induces human cancer cell death in vitro (v0.1)"

Structure–Function Studies and Mechanism of Action of Snake Venom L-Amino Acid Oxidases

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