Isolation and characterization of an N-acetyl-D-galactosamine-binding lectin from Dutch Iris bulbs which recognizes the blood group A disaccharide (GalNAc alpha 1-3Gal).
A blood group type A disaccharide (GalNAc alpha 1-3Gal)-binding lectin has been purified from Dutch Iris (Iris x hollandica) bulbs to electrophoretic homogeneity by consecutive affinity chromatography on columns of immobilized asialofetuin-Sepharose 4B and Synsorb A disaccharide (GalNAc alpha 1-3Gal beta-O-(CH2)8CONH-Synsorb). This lectin agglutinates both native and trypsin-treated rabbit erythrocytes, but not human erythrocytes, irrespective of blood group type. Gel filtration chromatography on Sephacryl S-200 HR column and SDS-polyacrylamide gel electrophoresis revealed the lectin to be a heterodimer consisting of two peptide chains (27 and 34 kDa) linked by disulfide bonds. The carbohydrate binding specificity of the lectin was investigated by quantitative precipitation, hemagglutination inhibition, and precipitation inhibition assays. It is a Gal/GalNAc-specific lectin, with an extended carbohydrate combining site, which appears to be most complementary to GalNAc linked to the C-3 or C-6 hydroxyl group of galactose. As inhibitors, these disaccharides are approximately 30-60 times more potent than galactose and 4-8-fold more active than N-acetyl-D-galactosamine, whereas both the blood type A trisaccharide (GalNAc alpha 1-3[L-Fuc alpha 1-2]Gal) and the Forssman disaccharide (GalNAc alpha 1-3GalNAc) are noninhibitory, suggesting the importance of a free equatorial hydroxyl group at the C-2 position of the penultimate galactose for lectin binding; either an acetamido group or a fucosyl group at this position appears to cause steric hindrance, thus abolishing binding to the lectin.
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Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
Van Damme
E J
EJ
Peumans
W J
WJ
Goldstein
I J
IJ
D000017:ABO Blood-Group System; D000116:Acetylgalactosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002240:Carbohydrate Sequence; D002241:Carbohydrates; D011232:Chemical Precipitation; D004187:Disaccharides / Q000378:metabolism*; D006241:Haptens; D006386:Hemagglutination Tests; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry*; D011817:Rabbits; D054790:Ribosome Inactivating Proteins, Type 2; D017386:Sequence Homology, Amino Acid
D000017:ABO Blood-Group System; D000596:Amino Acids; D002241:Carbohydrates; D004187:Disaccharides; D006241:Haptens; C086197:Iris lectin; D037102:Lectins; D037121:Plant Lectins; D054790:Ribosome Inactivating Proteins, Type 2; C086195:N-acetylgalactosaminyl-alpha(1-3)galactose; D000116:Acetylgalactosamine