Purification and characterization of a galactose-specific lectin with mitogenic activity from pinto beans

• Published in: Biochimica et biophysica acta Vol. 1760; no. 5; pp. 808 - 813
• Main Authors: Wong, Jack H., Wong, Clarence C.T., Ng, T.B.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2006
• Subjects: Animals; Cell Proliferation - drug effects; Chromatography; Galectins - isolation & purification; Galectins - pharmacology; HIV Reverse Transcriptase - antagonists & inhibitors; Hydrogen-Ion Concentration; Inhibitory Concentration 50; Isolation; Lectin; Leukemia L1210; Mice; Mitogens - isolation & purification; Mitogens - pharmacology; Molecular Weight; Phaseolus - chemistry; Pinto bean; Plant Lectins - isolation & purification; Plant Lectins - pharmacology; Spleen - cytology; Spleen - drug effects; Temperature; Lectin; Isolation; Pinto bean
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2006.02.015

A galactose-specific dimeric lectin from pinto beans was purified using a procedure that involved affinity chromatography on Affi-gel blue gel, anion exchange chromatography on Q-Sepharose, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 200. The molecular mass of this homodimeric lectin was 62 kDa and that of each of its subunits was 31 kDa. The hemagglutinating activity of pinto bean lectin was stable within the pH range of 3–12 and the temperature range of 0–70 °C. By using the [ 3H-methyl]-thymidine incorporation assay, it was shown that the lectin had the ability to evoke a mitogenic response from murine splenocytes but it did not inhibit proliferation of L1210 leukemia cells. The pinto bean lectin inhibited HIV-1 reverse transcriptase with an IC 50 of 3 μM.