Characterization of the carbohydrate binding specificity of the leukoagglutinating lectin from Maackia amurensis. Comparison with other sialic acid-specific lectins.
The sialic acid-specific leukoagglutinating lectin from the seeds of Maackia amurensis (MAL) has been studied by the techniques of quantitative precipitin formation, hapten inhibition of precipitation, hapten inhibition using an enzyme-linked immunosorbent assay, and lectin affinity chromatography. The ability of the immobilized lectin to fractionate oligosaccharides based on their content of sialic acid has also been investigated. Our results indicate that MAL reacts with greatest affinity with the trisaccharide sequence Neu5Ac/Gc alpha 2,3Gal beta 1,4GlcNAc/Glc. The lectin requires three intact sugar units for binding and does not interact when the beta 1,4-linkage is replaced by a beta 1,3-linkage nor when the "reducing sugar" of the trisaccharide is reduced. Results from enzyme-linked immunosorbent assays show that an N-acetyllactosamine repeating sequence is not required; however, the N-acetyllactosamine repeating sequence does appear to enhance the binding of MAL to a series of glycolipids. In addition, the sialic acid may be substituted with either N-acetyl or N-glycolyl groups without reduction in binding. The C-8 and C-9 hydroxyl groups of sialic acid do not play a role in binding as shown by the strong reaction of periodate-treated glycoproteins. Comparison of the specificity of the three sialic acid-binding lectins indicates that Limax flavus agglutinin binds to Neu5Ac in any linkage and in any position in a glycoconjugate, Sambucus nigra lectin requires a disaccharide of the structure Neu5Ac alpha 2,6Gal/GalNAc, and MAL has a binding site complimentary to the trisaccharide Neu5Ac alpha 2,3Gal beta 1,4GlcNAc/Glc, to which sialic acid contributes less to the total binding affinity than for either S. nigra lectin or L. flavus agglutinin.
Knibbs
R N
RN
Department of Biological Chemistry, University of Michigan Medical School, Ann Arbor 48109-0606.
Goldstein
I J
IJ
Ratcliffe
R M
RM
Shibuya
N
N
D000371:Agglutination; D000818:Animals; D002240:Carbohydrate Sequence; D002241:Carbohydrates; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D006001:Glycoconjugates; D006017:Glycolipids / Q000032:analysis; D006241:Haptens; D037102:Lectins; D008264:Macrophages / Q000276:immunology; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D012794:Sialic Acids
D002241:Carbohydrates; D006001:Glycoconjugates; D006017:Glycolipids; D006241:Haptens; D037102:Lectins; D012794:Sialic Acids