Abstract
A mannose-binding lectin (Narcissus tazetta lectin [NTL]) with potent antiviral activity was isolated and purified from the bulbs of the Chinese daffodil Narcissus tazetta var. chinensis, using ion exchange chromatography on diethylaminoethyl (DEAE)-cellulose, affinity chromatography on mannose-agarose and fast protein liquid chromatography (FPLC)-gel filtration on Superose 12. The purified lectin was shown to have an apparent molecular mass of 26 kDa by gel filtration and 13 kDa by SDS-PAGE, indicating that it is probably a dimer with two identical subunits. The cDNA-derived amino acid sequence of NTL as determined by molecular cloning also reveals that NTL protein contains a mature polypeptide consisting of 105 amino acids and a C-terminal peptide extension. Three-dimensional modelling study demonstrated that the NTL primary polypeptide contains three subdomains, each with a conserved mannose-binding site. It shows a high homology of about 60%–80% similarity with the existing monocot mannose-binding lectins. NTL could significantly inhibit plaque formation by the human respiratory syncytial virus (RSV) with an IC50 of 2.30 µg/ml and exhibit strong antiviral properties against influenza A (H1N1, H3N2, H5N1) and influenza B viruses with IC50 values ranging from 0.20 µg/ml to 1.33 µg/ml in a dose-dependent manner. It is worth noting that the modes of antiviral action of NTL against RSV and influenza A virus are significantly different. NTL is effective in the inhibition of RSV during the whole viral infection cycle, but the antiviral activity of NTL is mainly expressed at the early stage of the viral cycle of influenza A (H1N1) virus. NTL with a high selective index (SI=CC50/IC50≥141) resulting from its potent antiviral activity and low cytotoxicity demonstrates a potential for biotechnological development as an antiviral agent.
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Abbreviations
- 3D:
-
three-dimensional
- ATCC:
-
American Type Culture Collection
- CC:
-
cytotoxic concentration
- CoV:
-
coronavirus
- CPE:
-
cytopathic effect
- DEAE:
-
diethylaminoethyl
- DMSO:
-
dimethyl sulphoxide
- EMEM:
-
Eagle minimum essential medium
- FBS:
-
foetal bovine serum
- FPLC:
-
fast protein liquid chromatography
- GNA:
-
Galanthus nivalis agglutinin
- HA:
-
haemagglutination
- HCMV:
-
human cytomegalovirus
- HCV:
-
hepatitis C virus
- HEp2:
-
human larynx epidermoid carcinoma cell line
- HHA:
-
Hippeastrum hybrid agglutinin
- HIV:
-
human immunodeficiency virus
- HSV:
-
herpes simplex virus
- IC:
-
inhibitory concentration
- IFN:
-
interferon
- MDCK:
-
Madin-Darby canine kidney cell line
- MOI:
-
multiplicity of infection
- NTL:
-
Narcissus tazetta lectin
- OD:
-
optical density
- PBS:
-
phosphate-buffered saline
- RSV:
-
respiratory syncytial virus
- SCID:
-
severe combined immunodeficiency
- SDS:
-
sodium dodecyl sulphate
- SI:
-
selective index
- TGF:
-
transforming growth factor
- TPCK:
-
tolylsulphonyl phenylalanyl chloromethyl ketone
- UDA:
-
Urtica dioica agglutinin
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Ooi, L.S.M., Ho, WS., Ngai, K.L.K. et al. Narcissus tazetta lectin shows strong inhibitory effects against respiratory syncytial virus, influenza A (H1N1, H3N2, H5N1) and B viruses. J Biosci 35, 95–103 (2010). https://doi.org/10.1007/s12038-010-0012-8
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DOI: https://doi.org/10.1007/s12038-010-0012-8