Chemical and biological properties of a purified lectin from the seeds of Antipolo tree (Artocarpus blancoi Elm. Merr.)
2000
Alpante, B.M.
A non-blood type specific lectin was isolated from the seeds of Artocarpus blancoi (Elm.) Merr. by extraction using 0.02 M phosphate buffered saline in 0.15 M NaCl at pH 7.2, ammonium sulfate fractionation and gel chromatography using Sephadex G-200. The purity of the lectin was confirmed by polyacrylamide gel electrophoresis (PAGE) under non-denaturing conditions at pH 8.8.Gel chromatography showed that the lectin has an estimated molecular weight of 444 kD. SDS-PAGE revealed a single band corresponding to 234 kD, which indicates that the lectin is possibly a dimer. Temperatures greater than 80 deg C inactivated the lectin. The optimum temperature for its activity was 30-70 deg C. Chemical analysis and High Performance Liquid Chromatography revealed that the lectin is a glycoprotein with 0.119% total sugars. Mannose and galactose were identified to be present. The isolated lectin was mitogenic to human lymphocytes, and had significant toxic effects on brine shrimp (Artemia salina). It did not inhibit the growth of microorganisms such as Gram positive and negative bacteria, molds, and yeasts.MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphemyltetrazolium bromide) tests using lung cancel cells (A549) and Sarcoma 180 showed that the lectin did not kill the cells nor caused proliferation of the cell lines. However, increasing concentration of lectin lead to a decreasing trend in functional survival (FS) value of the cancer cells
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