Proteomic characterization of the thermostable toxins from Naja naja venom

Binh, DV; Thanh, TT; Chi, PV

doi:10.1590/s1678-91992010000400014

Cited by 11 publications

(7 citation statements)

References 25 publications

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“…Thus, similarly to the BCA method, the content of these amino acids in proteins will influence the results obtained. The Bradford assay has also been used many times to measure protein concentration in venom [8,29,[42][43][44][45]. Interestingly, this is the only method that indicates that the concentration of protein in A. contortrix venom is higher than in N. ashei and this result seems unlikely from our experience.…”

Section: Discussionmentioning

confidence: 90%

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Bocian

Sławek

Jaromin

et al. 2020

Animals

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Simple Summary: Snake venom is mostly composed of proteins and peptides, which are of interest to many researchers due to their potential pharmacological properties. Due to their biochemical character, these components are analyzed using proteomic techniques such as electrophoresis, chromatography and mass spectrometry. A very important stage of such studies is the measurement of protein concentration in the sample, which is most often performed by colorimetric methods. In the presented article, we used five such techniques on venoms of two snake species, namely Agkistrodon contortrix and Naja ashei. In the case of A. contortrix venom, four methods provide similar concentration values, whereas, in the case of N. ashei, the differences between results are very significant. The source of these differences should probably be seen in the differences in amino acid composition of proteins of these two venoms. With this report, we would like to draw attention to the need to select an appropriate method for measuring the concentration of protein in the venom, especially in the case of Elapid species.Abstract: Snake venom is an extremely interesting natural mixture of proteins and peptides, characterized by both high diversity and high pharmacological potential. Much attention has been paid to the study of venom composition of different species and also detailed analysis of the properties of individual components. Since proteins and peptides are the active ingredients in venom, rapidly developing proteomic techniques are used to analyze them. During such analyses, one of the routine operations is to measure the protein concentration in the sample. The aim of this study was to compare five methods used to measure protein content in venoms of two snake species: the Viperids representative, Agkistrodon contortrix, and the Elapids representative, Naja ashei. The study showed that for A. contortrix venom, the concentration of venom protein measured by four methods is very similar and only the NanoDrop method clearly stands out from the rest. However, in the case of N. ashei venom, each technique yields significantly different results. We hope that this report will help to draw attention to the problem of measuring protein concentration, especially in such a complex mixture as animal venoms.

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Section: Discussionmentioning

confidence: 90%

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Bocian

Sławek

Jaromin

et al. 2020

Animals

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“…Although reducing conditions were used for 2D-PAGE, subunits, monomeric forms, and/or native multimeric proteins that remained unreduced may present in these spots. Earlier studies have shown that high-molecular-weight snake venom proteins are more affected by heat than low-MW proteins and peptides [28][29][30]. LAAO isolated from Macrovipera lebetinus venom was also found to be stable from 4 to 25°C while heating at 70°C inactivated the enzyme in 15 min [27].…”

Section: Discussionmentioning

confidence: 94%

Effect of Temperature on the Protein Profile of Macrovipera Lebetinus (Blunt-Nosed Viper) Venom: A Preliminary Study

İĞCİ

2023

Journal of Scientific Reports-A

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Snake venom is a complex biological fluid consisting mainly of proteins and peptides possessing diverse biological activities. Snake venoms draw attention due to their bioactive proteins/peptides with therapeutic and diagnostic potential. Testing the stability of snake venom proteins under different conditions including temperature provides useful data for venom research. Macrovipera lebetinus, blunt-nosed viper, is the biggest viper species of Türkiye distributed mainly in eastern and southeastern Anatolia. Although its venom components were investigated before, there is limited data regarding the effect of temperature on its venom proteins. The present study aimed to investigate the effect of temperature on the venom proteins of M. lebetinus. For this purpose, venom samples were incubated at 25, 37, and 50°C. Thereafter, venom proteins were separated by two-dimensional gel electrophoresis (2D-PAGE) method. Some qualitative and quantitative differences in the protein profile indicating structural changes and degradation were observed especially after 50°C treatment. It has been found that the protein spots most affected by temperature will most likely contain metalloproteinase, phospholipase A2 and ʟ-amino acid oxidase enzymes, by comparing the experimental molecular weight and pI values with those in the literature. Detailed studies including enzyme activities and toxicity assays will provide more data on the stability of M. lebetinus venom under different conditions.

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“…In-gel digestion of proteins isolated by gel electrophoresis was carried out according to the protocol published by Shevchenko et al [25] with some modifications described in our previous study [3,28,29]. All chemicals including DTT, iodoacetamide (IAA), ammonium bicarbonate, ammonium acetate, trypsin (proteomics sequencing grade), sodium bicarbonate and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA) prepared using deionized filter-sterilised MilliQ water.…”

Section: In-gel Digestionmentioning

confidence: 99%