Phospholipase A2 enhances the endothelial cell detachment effect of a snake venom metalloproteinase in the absence of catalysis

Bustillo, Soledad; García-Denegri, María Emilia; Gay, Carolina; Velde, Andrea Carolina Van de; Acosta, Ofelia; Angulo, Yamileth; Lomonte, Bruno; Gutiérrez, José Marı́a; Leiva, Laura

doi:10.1016/j.cbi.2015.08.002

Cited by 36 publications

(23 citation statements)

References 37 publications

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“…Longer times of incubation enhanced B. lanceolatus venom cytotoxicity. Interestingly B. lanceolatus venom results reproduced the in vivo observations of Jiménez et al [ 45 ]; in a murine model of cutaneous tissue damage, class I SVMP, BaP1, causes the apoptosis of keratinocytes without affecting the viability of endothelial cells. As EAhy926 cells present more chromosomes than human umbilical vein endothelial cell (HUVECs), which could influence cells’ resistance to venom exposure, these results could be confirmed using HUVECs [ 25 , 48 ].…”

Section: Discussionsupporting

confidence: 82%

“…Several studies addressing the cytotoxic action of venoms of the Bothrops species from South America or the cytotoxicity of proteins purified from these venoms for endothelial cells have been performed [ 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 ], but there are few reports of Bothrops venom toxicity for keratinocytes [ 46 , 47 ]. To date, nothing is known about the cytotoxic potential of B. lanceolatus venom.…”

Section: Discussionmentioning

confidence: 99%

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Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation

Delafontaine

Villas-Boas

Mathieu

et al. 2017

Toxins

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Bothrops lanceolatus, commonly named ‘Fer-de-Lance’, is an endemic snake of the French Caribbean Island of Martinique. Envenomations by B. lanceolatus present clinical aspects characterized by systemic thrombotic syndrome and important local inflammation, involving edema and pain but limited hemorrhage. To investigate mechanisms of venom-induced inflammation, B. lanceolatus venom was characterized, its cross-reactivity with bothropic antivenom explored, its cytotoxicity on human keratinocytes and vascular cells, and the production of cytokines and chemokines were analyzed. We used electrophoretic separation, zymography, colorimetric or fluorimetric enzymatic assays, and immunochemical assays. Therapeutic South American bothropic antivenom cross-reacted with B. lanceolatus venom and completely or partially abolished its PLA2, hyaluronidase, and proteolytic activities, as well as its cytotoxicity for keratinocytes. The substrate specificity of B. lanceolatus venom proteases was emphasized. B. lanceolatus venom cytotoxicity was compared to the B. jararaca venom. Both venoms were highly cytotoxic for keratinocytes (HaCaT), whereas B. lanceolatus venom showed particularly low toxicity for endothelial cells (EAhy926). Patterns of cytokine and chemokine production by cells exposed to the venoms were highly pro-inflammatory. Thus, the results presented here show that B. lanceolatus venom toxins share important antigenic similarities with South American Bothrops species toxins, although their proteases have acquired particular substrate specificity. Moreover, the venom displays important cytotoxic and pro-inflammatory action on human cell types such as keratinocytes and endothelial cells, which are important players in the local and systemic compartments affected by the envenomation.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation

Delafontaine

Villas-Boas

Mathieu

et al. 2017

Toxins

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“…This is consistent with the report of non-toxic property of various acidic-type PLA 2 s isolated from the venoms of Indian N. kaouthia (Joubert & Taljaard, 1980) and Pakistani N. naja (Wong, Tan & Tan, 2016), but diverged markedly from the highly lethal, neutral/basic venom PLA 2 s characterised for other Southeast Asian cobras including Naja sumatrana and N. sputatrix (Leong et al, 2015; Tan & Arunmozhiarasi, 1989). The acidic PLA 2 s hence may serve a secondary role of ancillary function, for instance, potentiating the toxic actions of other venom components, including CTXs/cardiotoxins (Gasanov, Dagda & Rael, 2014), SVMPs (Bustillo et al, 2015) and weak neurotoxin (Mukherjee, 2010) to enhance tissue damages which are crucial for prey digestion but clinically deleterious as it complicates local tissue necrosis. On the other hand, although the mRNA level of the predominant form of PLA 2 was relatively low (∼2%) within the venom gland, the PLA 2 protein content in N. kaouthia venom is disproportionately higher (12–14% of total venom proteins) (Laustsen et al, 2015; Tan et al, 2015d).…”

Section: Resultsmentioning

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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“…Cytolysis was assessed by monitoring the release of the cytosolic marker enzyme lactate dehydrogenase (LDH) using a commercial kit (Wiener ® , LDH-P UV, Buenos Aires, Argentina), as previously described [30]. Reference controls for 0% and 100% cytolysis consisted of medium alone and medium from cells incubated with 0.1% Triton X-100 (v/v), respectively.…”

Section: Cytotoxicity and Morphological Analysis Cytotoxicity Assaymentioning

confidence: 99%

Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)

Fusco

Cajade

Piñeiro

et al. 2020

J. Venom. Anim. Toxins incl. Trop. Dis

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Background: Argenteohyla siemersi (red-spotted Argentina frog) is a casque-headed tree frog species belonging to the Hylidae family. This species has a complex combination of anti-predator defense mechanisms that include a highly lethal skin secretion. However, biochemical composition and biological effects of this secretion have not yet been studied. Methods: The A. siemersi skin secretion samples were analyzed by mass spectrometry and chromatographic analysis (MALDI-TOF/MS, RP-HPLC and GC-MS). Proteins were also studied by SDS-PAGE. Among the biological activities evaluated, several enzymatic activities (hemolytic, phospholipase A 2 , clotting, proteolytic and amidolytic) were assessed. Furthermore, the cytotoxic activity (cytolysis and fluorescence staining) was evaluated on myoblasts of the C2C12 cell line. Results: The MALDI-TOF/MS analysis identified polypeptides and proteins in the aqueous solution of A. siemersi skin secretion. SDS-PAGE revealed the presence of proteins with molecular masses from 15 to 55 kDa. Steroids, but no alkaloids or peptides (less than 5 kDa), were detected using mass spectrometry. Skin secretion revealed the presence of lipids in methanolic extract, as analyzed by CG-MS. This secretion showed hemolytic and phospholipase A 2 activities, but was devoid of amidolytic, proteolytic or clotting activities. Moreover, dose-dependent cytotoxicity in cultured C2C12 myoblasts of the skin secretion was demonstrated. Morphological analysis, quantification of lactate dehydrogenase release and fluorescence staining indicated that the cell death triggered by this secretion involved necrosis. Conclusions: Results presented herein evidence the biochemical composition and biological effects of A. siemersi skin secretion and contribute to the knowledge on the defense mechanisms of casque-headed frogs.

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Phospholipase A2 enhances the endothelial cell detachment effect of a snake venom metalloproteinase in the absence of catalysis

Cited by 36 publications

References 37 publications

Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation

Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation

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

Biochemical characterization and cytotoxic effect of the skin secretion from the red-spotted Argentina frog Argenteohyla siemersi (Anura: Hylidae)

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