Separation of oligosaccharides containing terminal alpha-linked galactose residues by affinity chromatography on Griffonia simplicifolia I bound to concanavalin A-sepharose.
The seeds of Griffonia simplicifolia contain a family of five isolectins (GS-I) (L. A. Murphy and I. J. Goldstein (1977) J. Biol. Chem. 252, 4739-4742) that bind with high affinity to glycoconjugates containing terminal nonreducing alpha-linked galactose residues. Here, we report that GS-I itself is bound via its high mannose-type, Asn-linked sugar chains to immobilized concanavalin A (Con A-Sepharose). The GS-I in the GS-I-Con A-Sepharose complex retains its ability to bind glycoconjugates containing terminal alpha-linked galactose residues. This convenient method to immobilize GS-I is rapid and quantitative. We have exploited this affinity system to separate oligosaccharides based on their number of terminal alpha-linked D-galactose residues.
Wang
W C
WC
Department of Biochemistry School of Chemical Sciences, University of Georgia, Athens 30602.
Clark
G F
GF
Smith
D F
DF
Cummings
R D
RD
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004912:Erythrocytes / Q000032:analysis; D005690:Galactose; D006017:Glycolipids / Q000097:blood / Q000302:isolation & purification; D037102:Lectins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D037121:Plant Lectins; D011817:Rabbits; D012685:Sepharose / Q000031:analogs & derivatives
D006017:Glycolipids; C045475:Griffonia simplicifolia lectins; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins; C028326:concanavalin A-sepharose; D012685:Sepharose; D005690:Galactose