Purification and characterization of the alpha-1,3-mannosylmannose-recognizing lectin of Crocus vernus bulbs.
A unique mannose-binding lectin, highly specific for terminal Man(alpha1,3)Man groups, was isolated from bulbs of crocus (Crocus vernus All.). The lectin failed to bind to a mannose affinity column and was purified by simple gel permeation chromatography (Sephacryl S200). The purified lectin, obtained in crystalline form, had a molecular mass of 44 kDa on gel filtration and showed a single peptide band with a molecular mass of 11 kDa on SDS-polyacrylamide gel electrophoresis, indicating it to be a tetrameric protein composed of four identical subunits. The N-terminal amino acid sequence analysis of the crocus lectin showed essentially no homology with that of other mannose-binding bulb lectins. The crocus lectin selectively interacted with the wild type Saccharomyces cerevisiae and other mannans carrying terminal Man(alpha1,3)Man but not with those lacking this disaccharide unit. In hapten inhibition studies, methyl alpha-mannopyranoside did not inhibit the mannan-lectin interaction. Of various alpha-mannooligosaccharides, those having the Man(alpha1,3)Man sequence showed the highest inhibitory potency, confirming the strict requirement of lectin for terminal alpha1,3-linked mannosylmannose units. An affinity column of immobilized lectin enabled the complete resolution of yeast mannan and glycogen. The immobilized lectin may provide a useful tool for purification and analysis of biologically important polysaccharides and glycoproteins.
Misaki
A
A
Konan Women's University, 6-2-23 Morikita-machi, Higasinada, Kobe 658, Japan.
Kakuta
M
M
Meah
Y
Y
Goldstein
I J
IJ
D002240:Carbohydrate Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D003079:Colchicum / Q000737:chemistry*; D004187:Disaccharides / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification*; D008351:Mannans / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D037121:Plant Lectins; D010946:Plants, Medicinal; D011134:Polysaccharides / Q000378:metabolism; D012441:Saccharomyces cerevisiae
D002352:Carrier Proteins; D004187:Disaccharides; D006023:Glycoproteins; D037102:Lectins; D008351:Mannans; D037121:Plant Lectins; D011134:Polysaccharides; C108662:1,3-mannosylmannose