Purification and Characterization of a Lectin from Phaseolus vulgaris cv. (Anasazi Beans)

A lectin has been isolated from seeds of the Phaseolus vulgaris cv. Anasazi beans using a procedure that involved affinity chromatography on A.-gel blue gel, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S, and FPLC-gel filtration on Superdex 200. The lectin was comprised of two 30-kDa subunits with substantial N-terminal sequence similarity to other Phaseolus lectins. The hemagglutinating activity of the lectin was stable within the pH range of 1-14 and the temperature range of 0-80 degrees C. The lectin potently suppressed proliferation of MCF-7 (breast cancer) cells with an IC50 of 1.3 mu M, and inhibited the activity of HIV-1 reverse transcriptase with an IC50 of 7.6 mu M. The lectin evoked a mitogenic response from murine splenocytes as evidenced by an increase in [H-3-methyl]-thymidine incorporation. The lectin had no antifungal activity. It did not stimulate nitric oxide production by murine peritoneal macrophages. Chemical modification results indicated that tryptophan was crucial for the hemagglutinating activity of the lectin. Copyright (C) 2009 Arishya Sharma et al.