Abstract

The lectin found in the tubers of the Winter Aconite (Eranthis hyemalis) plant (EHL) is a Type II Ribosome Inactivating Protein (RIP). Type II RIPs have shown anti-cancer properties and have great potential as therapeutic agents. Similarly, colloidal gold nanoparticles are successfully used in biomedical applications as they can be functionalised with ligands with high affinity and specificity for target cells to create therapeutic and imaging agents. Here we present the synthesis and characterization of gold nanoparticles conjugated with EHL and the results of a set of initial assays to establish whether the biological effect of EHL is altered by the conjugation. Gold nanoparticles functionalised with EHL (AuNPs@EHL) were successfully synthesised by bioconjugation with citrate gold nanoparticles (AuNPs@Citrate). The conjugates were analysed by UV-Vis spectroscopy, Dynamic Light Scattering (DLS), Zeta Potential analysis, and Transmission Electron Microscopy (TEM). Results indicate that an optimal functionalisation was achieved with the addition of 100 µL of EHL (concentration 1090 ± 40 µg/mL) over 5 mL of AuNPs (concentration [Au0] = 0.8 mM). Biological assays on the effect of AuNPs@EHL were undertaken on Caenorhabditis elegans, a free-living nematode commonly used for toxicological studies, that has previously been shown to be strongly affected by EHL. Citrate gold nanoparticles did not have any obvious effect on the nematodes. For first larval stage (L1) nematodes, AuNPs@EHL showed a lower biological effect than EHL. For L4 stage, pre-adult nematodes, both EHL alone and AuNPs@EHL delayed the onset of reproduction and reduced fecundity. These assays indicate that EHL can be conjugated to gold nanoparticles and retain elements of biocidal activity.

Highlights

  • Lectins are a class of proteins ubiquitously expressed in plants, animals, bacteria, and viruses.They are well known for their ability to agglutinate erythrocytes, and their ability to bind carbohydrates selectively based on the individual sugar specificity of the lectin [1,2]

  • The particles were analyzed by UV-Vis spectroscopy (JASCO Co., Tokyo, Japan), Dynamic Light Scattering (DLS) (MALVERN, Panalytical, UK), Zeta Potential (MALVERN, Panalytical, UK), and Transmission Electron Microscopy (TEM) (JEM 1010, JEOL, Tokyo, Japan) analysis

  • The synthesis and characterization of gold nanoparticles conjugated with EHL (AuNPs@EHL)

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Summary

Introduction

Lectins are a class of proteins ubiquitously expressed in plants, animals, bacteria, and viruses. They are well known for their ability to agglutinate erythrocytes, and their ability to bind carbohydrates selectively based on the individual sugar specificity of the lectin [1,2]. Eranthis hyemalis (Winter Aconite) is a late winter/early spring flowering perennial plant of the family Ranunculaceae. E. hyemalis possesses a proteinaceous toxin (named Eranthis hyemalis lectin, EHL), found to cause agglutination of erythrocytes as well as impacting on the fitness of some agricultural pests and plant viruses [5,9]. E. hyemalis is the sole representative of the Ranunculaceae to be reported to express lectin activity. Due to the structural and toxicity studies conducted [5,9,10,11], EHL should be classed as a Type II Ribosome Inactivating Protein (RIP)

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