Purification, characterization and toxicity of a mannose-binding lectin from the seeds of Treculia africana plant

Abstract

In this study, a mannose-specific, homodimeric lectin from the seeds of Treculia africana was purified, characterized and its adverse effects were investigated in mice. The purification protocol involved anionic exchange chromatography on DEAE-Cellulose followed by gel filtration on Sephadex G-100. The hemagglutinating activity of lectin towards human erythrocytes was sensitive to inhibition by D-mannose. Treatment of the protein with EDTA exerted no inhibitory effect; however, analysis of metal content by atomic absorption spectroscopy revealed the presence of Cu2+, Fe3+, and Mg2+. The results obtained showed that the lectin possesses maximum hemagglutinating activity towards human erythrocytes activity over the pH range 3–7.2 and is relatively thermostable up to 50°C. Periodic acid Schiff's (PAS) reagent staining showed that the protein was non-glycosylated while its amino acid composition analysis revealed that the protein contained 155 residues per subunit. The subunit had a minimal molecular weight of 22,139 Daltons, while the native molecular weight was estimated to be 41,000 Daltons. The lectin was found to be moderately toxic to mice with an LD50 of 47.21 µg g−1 body weight while, histopathological analysis showed no treatment related effects in any of the organs examined.