Studies on phytohemagglutinins XXVIII. Chemical modifications of the lectin from seeds of the lentil ( lens esculenta moench.)

Abstract

The reaction of lentil lectin with maleic anhydride yields maleyl derivatives with a degree of modification depending on the amount of used maleic anhydride. Hemagglutinating activity of maleyl derivatives decreases with the increasing number of modified lysine residues, but even at a maleylation of 90% of free -NH 2 groups, the maleyl derivative of the lectin still retains a hemagglutinating activity. Maleylation of the lentil lectin is not accompanied by dissociation; the chemically modified lectin also retains its bound Mn. Maleyl derivatives of the lectin retain the ability to interact specifically with polysaccharides (Sephadex, mannan). After removal of maleyl groups in acidic medium, the hemagglutinating activity does not differ from that of the native lectin. Acetylation of the lentil lectin with N- acetylimidazole results in formation of a modified protein containing 3–5 acetylated -OH groups of the tyrosyl residues out of the total of 15 groups and 7–17 acetylated -NH 2 groups out of the total of 26 free -NH 2 groups, depending upon the reaction conditions. The presence of d-mannose in the reaction mixture does not affect the extent of modification of tyrosyl groups, but it decreases the modification of -NH 2 groups. Acetyl derivatives of the lentil lectin do not possess hemagglutinating activity, but they retain the ability to interact with polysaccharides and to bind to erythrocytes. Acetylation does not affect the content of protein bound Mn. After de-O-acetylation, the hemagglutinating activity of the modified protein increases to the value of the native protein.