Abstract
A 64-kDa dimeric lectin was purified from Phaseolus vulgaris cv. Japanese mottled beans. The purification protocol involved ion exchange chromatography with Q-Sepharose and SP-Sepharose and size exclusion chromatography on Superdex 75. The lectin was adsorbed on both Q-Sepharose and SP-Sepharose columns. Finally, the lectin gave a sharp absorbance peak which corresponded to 64 kDa based on results of size exclusion chromatography. Sodium dodecyl sulphate- polyacrylamide gel electrophoresis displayed a single band at around 32 kDa, indicating that the protein was dimeric. The hemagglutination inhibition assay indicated that the lectin showed specificity toward galactose. The lectin preserved hemagglutinating activity below 70 °C and at a pH range 3 – 12. The lectin was able to inhibit proliferation of MCF-7 cells and Hep G2 cells and possessed antifungal activity toward Mycosphaeralla arachidicola with an IC50 value of 3.9 µM. The activity of HIV-1 reverse transcriptase was reduced by 61.9 % in the presence of the lectin at 6.25 µM concentration.
Keywords: Antitumor, Japanese mottled beans, lectin, purification.
Protein & Peptide Letters
Title:Isolation and Characterization of a Lectin from Japanese Mottled Beans
Volume: 21 Issue: 7
Author(s): Yuan Zhao, Ameer Maqsood Ahmad, Randy Chi Fai Cheung and Tzi Bun Ng
Affiliation:
Keywords: Antitumor, Japanese mottled beans, lectin, purification.
Abstract: A 64-kDa dimeric lectin was purified from Phaseolus vulgaris cv. Japanese mottled beans. The purification protocol involved ion exchange chromatography with Q-Sepharose and SP-Sepharose and size exclusion chromatography on Superdex 75. The lectin was adsorbed on both Q-Sepharose and SP-Sepharose columns. Finally, the lectin gave a sharp absorbance peak which corresponded to 64 kDa based on results of size exclusion chromatography. Sodium dodecyl sulphate- polyacrylamide gel electrophoresis displayed a single band at around 32 kDa, indicating that the protein was dimeric. The hemagglutination inhibition assay indicated that the lectin showed specificity toward galactose. The lectin preserved hemagglutinating activity below 70 °C and at a pH range 3 – 12. The lectin was able to inhibit proliferation of MCF-7 cells and Hep G2 cells and possessed antifungal activity toward Mycosphaeralla arachidicola with an IC50 value of 3.9 µM. The activity of HIV-1 reverse transcriptase was reduced by 61.9 % in the presence of the lectin at 6.25 µM concentration.
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Cite this article as:
Zhao Yuan, Ahmad Maqsood Ameer, Cheung Chi Fai Randy and Ng Bun Tzi, Isolation and Characterization of a Lectin from Japanese Mottled Beans, Protein & Peptide Letters 2014; 21 (7) . https://dx.doi.org/10.2174/0929866521666140320100308
DOI https://dx.doi.org/10.2174/0929866521666140320100308 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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