Mannose-specific isolectins with different hemagglutinating potencies isolated from Chinese daffodil (Narcissus tazetta var. chinensis) leaves.
Three mannose-specific lectins exhibiting considerable similarities in NH2-terminal amino acid sequence were isolated from leaves of the Chinese daffodil Narcissus tazetta (Family Amaryllidaceae). The purification protocol involved extraction with an aqueous buffer, anion exchange chromatography on DEAE-cellulose using stepwise elution with increasing salt concentrations, affinity chromatography on mannose-agarose, and FPLC-gel filtration on Superose 12. From the peak unadsorbed on DEAE-cellulose, and two peaks adsorbed on the ion exchanger and eluted respectively with 0.2 M Tris-HCl buffer and 0.5 M NaCl, were prepared fractions which yielded isolectins 1, 2, and 3 after adsorption on mannose-agarose and FPLC-gel filtration. All three isolectins were homodimers with a molecular weight of 26 kDa. The lectin unadsorbed on DEAE-cellulose had the lowest, while the most strongly adsorbed lectin had the highest hemagglutinating activity.
Ooi
L S
LS
Department of Biochemistry, Chinese University of Hong Kong, Shatin, New Territories.
Ng
T B
TB
Sun
S S
SS
Ooi
V E
VE
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006384:Hemagglutination; D006386:Hemagglutination Tests / Q000379:methods; D037102:Lectins / Q000302:isolation & purification* / Q000378:metabolism; D008358:Mannose / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018515:Plant Leaves / Q000737:chemistry*; D037121:Plant Lectins; D010940:Plant Proteins / Q000302:isolation & purification* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D037102:Lectins; D037121:Plant Lectins; D010940:Plant Proteins; D008358:Mannose