Winter Aconite (Eranthis hyemalis) Lectin as a cytotoxic effector in the lifecycle ofCaenorhabditis elegans

Abstract

The lectin found in the tubers of the Winter Aconite (Eranthis hyemalis) plant is an N-acetyl-D-galactosamine specific Type II Ribosome Inactivating Protein (RIP); Type II RIPs have shown anti-cancer properties, and hence have potential as therapeutic agents. Here we present a modified protocol for the extraction and purification of the E. hyemalis lectin (EHL) using affinity chromatography. De novo amino acid sequencing of EHL confirms its classification as a Type II Ribosome Inactivating Protein. The biocidal properties of EHL have been investigated against the nematode Caenorhabditis elegans. Arrested first stage larvae treated with EHL have shown some direct mortality, with surviving larvae subsequently showing a range of phenotypes including food avoidance, reduced fecundity, developmental delay and constitutive dauer larvae formation. Both inappropriate dauer larvae development and failure to locate to bacterial food source are consistent with the disruption of chemosensory function and the ablation of amphid neurons. Further investigation indicates that mutations that disrupt normal amphid formation can block the EHL-induced dauer larvae formation. In combination, these phenotypes indicate that EHL is cytotoxic and suggest a cell specific activity against the amphid neurons of C. elegans.

Transcriptomic Analysis of Aedes aegypti in Response to Mosquitocidal Bacillus thuringiensis LLP29 Toxin

“…Lectins are a class of carbohydrate binding proteins ubiquitously expressed in plants, animals, bacteria, and viruses 62 . The lectin is a major constituent of humoral immune system of insects, and is involved in the process of self/non-self-recognition.…”

Synthesis of Gold Functionalised Nanoparticles with the Eranthis hyemalis Lectin and Preliminary Toxicological Studies on Caenorhabditis elegans

“…These EHL-induced dauer larvae were also unable to resume development when maintained on food. The authors called this a “lectin-induced effect” and suggested that the occurrence of dauer formation and a failure to recover in the presence of food indicates that EHL is binding specifically to amphid neurons [ 16 ].…”

“…Here, two sets of experiments were performed. The first set was a replication of the work on the effects of EHL on C. elegans [ 16 ], using AuNPs@EHL. The second set of experiments (set 2) were carried out using L4 stage (pre-adult) worms and had not previously been conducted with EHL.…”

“…Previous work by some of the authors has shown biocidal effects of EHL against the free-living nematode Caenorhabditis elegans [ 16 ]. C. elegans is a well-established model organism for initial toxicological studies due to the conserved nature of its biological and biochemical processes, including the stress response and disease pathways [ 17 ].…”

“…The conjugates were analysed by UV-Vis spectroscopy, Dynamic Light Scattering (DLS), Zeta Potential analysis, and Transmission Electron Microscopy (TEM). Biological assays on the effect of AuNPs@EHL on C. elegans were performed using first life stage (L1) and pre-adult stage (L4) nematodes, and the results compared to previously published data on the effect of EHL on L1 nematodes [ 16 ]. Additionally, the effects of naked AuNPs on L1 and L4 nematodes, and the effect of EHL on L4s were also studied.…”