Purification and partial characterization of a lectin from the fresh leaves of Kalanchoe crenata (Andr.) Haw.
A haemagglutinating protein from the saline extracts of Kalanchoe crenata leaves, which agglutinate all human blood types, was purified to homogeneity by ion-exchange chromatography on a DEAE-Cellulose column followed by gel filtration on a Sephadex G-100 column. The purified protein showed one band, both in non-denaturing PAGE and SDS-PAGE. The M(r) that was determined by SDS-PAGE was 44,000 Da and that estimated from gel filtration was 47,000. Treatment of the haemagglutinating protein with 5 mM EDTA diminished the haemagglutinating activity to 50% of the original level. The addition of divalent cations, 10 mM Mg(2+), 10 mM Mn(2+), or 10mM Ba(2+), totally restored and enhanced the activity. The protein showed maximum activity over the 3-7 pH range and was heat-resistant. It was also a glycoprotein containing about 1.5% carbohydrate.
Department of Biochemistry, Obafemi Awolowo University, Ile-Ife, Nigeria. takuku@oauife.edu.ng
Kuku
K
Adenike
Oladiran Babalola
OB
Eretan
D000925:Anticoagulants / Q000494:pharmacology; D001464:Barium / Q000494:pharmacology; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry; D006384:Hemagglutination; D006863:Hydrogen-Ion Concentration; D031962:Kalanchoe / Q000033:anatomy & histology*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D018515:Plant Leaves / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry* / Q000302:isolation & purification*; D013696:Temperature
D000925:Anticoagulants; D002241:Carbohydrates; D006023:Glycoproteins; D037121:Plant Lectins; D001464:Barium; D008345:Manganese; D004492:Edetic Acid; D008274:Magnesium