Comparative analysis of the venom proteome of four important Malaysian snake species

Vejayan, Jaya; Khoon, Too Lay; Ibrahim, Halijah

doi:10.1186/1678-9199-20-6

Cited by 22 publications

(28 citation statements)

References 35 publications

(41 reference statements)

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“…The current systematics based on molecular phylogenetics has resolved the issue and clearly regarded the species of monocled cobra as N. kaouthia (Wüster, 1996). The venom proteome of N. kaouthia had also been studied to some extents over the years (Kulkeaw et al, 2007; Laustsen et al, 2015; Vejayan, Khoon & Ibrahim, 2014). It is noteworthy that a recent study has provided a global comparison on the proteomic details of N. kaouthia venoms from three different Southeast Asian regions (Malaysia, Thailand and Vietnam) qualitatively (profiling protein subtypes) and quantitatively (addressing relative abundances of the proteins) (Tan et al, 2015d).…”

Section: Introductionmentioning

confidence: 99%

Comparative venom gland transcriptomics ofNaja kaouthia(monocled cobra) from Malaysia and Thailand: elucidating geographical venom variation and insights into sequence novelty

Tan

Chanhome

et al. 2017

PeerJ

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BackgroundThe monocled cobra (Naja kaouthia) is a medically important venomous snake in Southeast Asia. Its venom has been shown to vary geographically in relation to venom composition and neurotoxic activity, indicating vast diversity of the toxin genes within the species. To investigate the polygenic trait of the venom and its locale-specific variation, we profiled and compared the venom gland transcriptomes of N. kaouthia from Malaysia (NK-M) and Thailand (NK-T) applying next-generation sequencing (NGS) technology.MethodsThe transcriptomes were sequenced on the Illumina HiSeq platform, assembled and followed by transcript clustering and annotations for gene expression and function. Pairwise or multiple sequence alignments were conducted on the toxin genes expressed. Substitution rates were studied for the major toxins co-expressed in NK-M and NK-T.Results and discussionThe toxin transcripts showed high redundancy (41–82% of the total mRNA expression) and comprised 23 gene families expressed in NK-M and NK-T, respectively (22 gene families were co-expressed). Among the venom genes, three-finger toxins (3FTxs) predominated in the expression, with multiple sequences noted. Comparative analysis and selection study revealed that 3FTxs are genetically conserved between the geographical specimens whilst demonstrating distinct differential expression patterns, implying gene up-regulation for selected principal toxins, or alternatively, enhanced transcript degradation or lack of transcription of certain traits. One of the striking features that elucidates the inter-geographical venom variation is the up-regulation of α-neurotoxins (constitutes ∼80.0% of toxin’s fragments per kilobase of exon model per million mapped reads (FPKM)), particularly the long-chain α-elapitoxin-Nk2a (48.3%) in NK-T but only 1.7% was noted in NK-M. Instead, short neurotoxin isoforms were up-regulated in NK-M (46.4%). Another distinct transcriptional pattern observed is the exclusively and abundantly expressed cytotoxin CTX-3 in NK-T. The findings suggested correlation with the geographical variation in proteome and toxicity of the venom, and support the call for optimising antivenom production and use in the region. Besides, the current study uncovered full and partial sequences of numerous toxin genes from N. kaouthia which have not been reported hitherto; these include N. kaouthia-specific l-amino acid oxidase (LAAO), snake venom serine protease (SVSP), cystatin, acetylcholinesterase (AChE), hyaluronidase (HYA), waprin, phospholipase B (PLB), aminopeptidase (AP), neprilysin, etc. Taken together, the findings further enrich the snake toxin database and provide deeper insights into the genetic diversity of cobra venom toxins.

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Section: Introductionmentioning

confidence: 99%

Comparative venom gland transcriptomics ofNaja kaouthia(monocled cobra) from Malaysia and Thailand: elucidating geographical venom variation and insights into sequence novelty

Tan

Chanhome

et al. 2017

PeerJ

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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: 89%

“…As such, venom components are of wide interest to many research groups and have been the subject of extensive research. As about 90% of venom components are of peptide origin, all venom analyses use increasingly fast developing proteomic techniques, including electrophoresis [8][9][10][11], immunodetection techniques [12][13][14][15], different types of chromatography [16][17][18][19] and mass spectrometry [20][21][22][23]. Each proteomic method has specific requirements for both the sample components and the protein concentration range.…”

Section: Introductionmentioning

confidence: 99%

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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“…A peçonha apresentou-se como uma fonte rica em fosfolipases A2, desintegrinas, lectina do tipo C, metaloproteinase, L-aminoácido oxidase e serinoprotease (VEJAYAN et al, 2014).…”

Section: L-aminoácido Oxidase De Calloselasma Rhodostoma (Cr-laao)unclassified

Caracterização funcional e estudo dos mecanismos de resposta ao dano celular causado por uma L-aminoácido oxidase de Calloselasma rhodostoma em linhagens celulares humanas

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As L-aminoácido oxidases (LAAOs) isoladas de peçonhas de serpentes são alvos de um grande número de pesquisas devido às suas inúmeras ações biológicas e farmacológicas. O objetivo do presente trabalho foi caracterizar funcionalmente a Laminoácido oxidase da peçonha de Calloselasma rhodostoma (CR-LAAO) por meio das atividades: bactericida, fungicida, leishmanicida, tripanomicida, citotóxica, inflamatória e análise da expressão de genes e proteínas apoptóticas. A CR-LAAO mostrou-se altamente citotóxica sobre as células tumorais HepG2 e HL-60, promovendo cerca de 80% de morte celular na maior concentração testada (100 µg/mL) e apresentou baixa toxicidade sobre células PBMC. Foi possível observar que a proteína induziu apoptose (AV+) em PBMC. Em HepG2, as menores concentrações (0,1-2,5 µg/mL) causaram apoptose (AV+), e as maiores (5-100 µg/mL) causaram apoptose/necrose (PI+/AV+). Em HL-60, as concentrações testadas (0,1-100 µg/mL) induziram apoptose/necrose (PI+/AV+). A expressão do gene FAS e ativação das caspases 8 e 3 determinou a ativação da via extrínseca da apoptose na linhagem HL-60. A CR-LAAO promoveu algumas alterações na modulação do ciclo celular, sendo que nas linhagens tumorais os atrasos se concentraram nas fases G0/G1 e S do ciclo celular. Ademais, a CR-LAAO mostrouse bactericida contra as cepas S. aureus e E. coli, com maior especificidade para a cepa gram-positiva (S. aureus). Em análises de microscopia de transmissão, foi possível observar um desmantelamento da parede celular bacteriana. Após 6h de pré-incubação com C. albicans, a CR-LAAO foi capaz de inibir 80% do crescimento da levedura. A CR-LAAO mostrou-se também um bom agente leishmanicida contra as espécies L. infantum chagasi (IC50=16,66 µg/mL) e L. braziliensis (IC50=24,47 µg/mL) e inibiu o crescimento da forma promastigota do Trypanosoma cruzi (IC50=196,8 µg/mL). Testes in vivo revelaram que a CR-LAAO promoveu inflamação local aguda, recrutando células inflamatórias como neutrófilos e induzindo a formação de citocinas (IL-6 e IL-1β) e mediadores lipídicos (LTB4 e PGE2). Os resultados obtidos sugerem que a CR-LAAO apresenta potencial biotecnológico evidente, com efeitos antiparasitários, fungicida, bactericida, bem como atividade antitumoral in vitro. Dessa forma, os resultados obtidos para a CR-LAAO fornecem subsídios importantes para o desenvolvimento de estratégias terapêuticas de ação direcionada, como quimioterápicos e antimicrobianos mais eficazes.Palavras chaves: Calloselasma rhodostoma; L-aminoácido oxidase; apoptose; expressão gênica; atividade antiparasitária; bactericida; inflamação. L-amino acid oxidases (LAAOs) isolated from snake venoms are targets of a large number of researches due to their numerous biological and pharmacological actions. The objective of this study was to functionally characterize the L-amino acid oxidase from the venom of Calloselasma rhodostoma (CR-LAAO) through the activities: bactericidal, fungicidal, leishmanicidal, trypanocidal, cytotoxic, inflammatory and analysis of gene expression and apoptotic pro...

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Comparative analysis of the venom proteome of four important Malaysian snake species

Cited by 22 publications

References 35 publications

Comparative venom gland transcriptomics ofNaja kaouthia(monocled cobra) from Malaysia and Thailand: elucidating geographical venom variation and insights into sequence novelty

Comparative venom gland transcriptomics ofNaja kaouthia(monocled cobra) from Malaysia and Thailand: elucidating geographical venom variation and insights into sequence novelty

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Caracterização funcional e estudo dos mecanismos de resposta ao dano celular causado por uma L-aminoácido oxidase de Calloselasma rhodostoma em linhagens celulares humanas

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