Effects of 3FTx Protein Fraction from Naja ashei Venom on the Model and Native Membranes: Recognition and Implications for the Mechanisms of Toxicity

Abstract

Three-finger toxins are naturally occurring proteins in Elapidae snake venoms. Nowadays, they are gaining popularity because of their therapeutic potential. On the other hand, these proteins may cause undesirable reactions inside the body′s cells. A full assessment of the safety of Naja ashei venom components for human cell application is still unknown. The aim of the study was to determine the effect of the exogenous application of three-finger toxins on the cells of monocytes (U-937) and promyelocytes (HL-60), with particular emphasis on the modification of their membranes under the influence of various doses of 3FTx protein fraction (0–120 ng/mL). The fraction exhibiting the highest proportion of 3FTx proteins after size exclusion chromatography (SEC) separation was used in the experiments. The structural response of cell membranes was described on the basis of single-component and multi-component Langmuir monolayers that mimicked the native membranes. The results show that the mechanism of protein–lipid interactions depends on both the presence of lipid polar parts (especially zwitterionic type of lipids) and the degree of membrane saturation (the greatest-for unsaturated lipids). The biochemical indicators reflecting the tested cells (MDA, LDH, cell survival, induction of inflammation, LD50) proved the results that were obtained for the model.

Development of an Inhibition Enzyme-Linked Immunosorbent Assay (ELISA) Prototype for Detecting Cytotoxic Three-Finger Toxins (3FTxs) in African Spitting Cobra Venoms

“…The venom was reconstituted in distilled water and stored at −20 °C until use. Three-finger toxin (3FTx) fraction was purified using size-exclusion chromatography as described elsewhere [ 39 ]. Venoms from Naja nigricollis , Naja haje , Bitis arietans , and Dendroaspis polylepis were obtained from Baringo Snake Park, Kenya, in liquid form.…”

Exploring nature’s antidote: unveiling the inhibitory potential of selected medicinal plants from Kisumu, Kenya against venom from some snakes of medical significance in sub-Saharan Africa

“…B. arietans, N. ashei, and N. subfulva are snakes of medical importance in sub-Saharan Africa (Calvete et al, 2007;Currier, 2012;Tasoulis and Isbister, 2017;Onyango, 2018;Okumu et al, 2020;Dyba et al, 2021) (Figure 1). Antivenom is the mainstay of treatment for envenomation by these snakes but is expensive, has limited availability, and does not sufficiently neutralize some key venom toxins, e.g., cytotoxins which cause dermonecrosis in snake bite victims.…”

Preclinical efficacy testing of three antivenoms against Naja ashei venom-induced lethality

“…A PLA 2 and 3FTx rich protein fraction of NAV showed good antimicrobial activity against Staphylococcus epidermidis ( Bocian et al, 2020a ). Moreover, a study on the exogenous application of 3FTx from NAV on monocytes (U-937) and pro myelocytes (HL-60) and their effect on cell membranes reported that the mechanism of protein-lipid interactions depends on the presence of lipid polar parts and the degree of membrane saturation ( Dyba et al, 2021 ). Bocian and colleagues compared the protein concentration of venoms from Naja ashei and Agkistrodon contortix using the Pierce™BCA Protein Assay Kit, the 2-D Quant Kit, the Qubit® Protein Assay Kit, the Bradford Assay, and the Nanodrop method ( Bocian et al, 2020b ).…”