The interaction of elderberry (Sambucus sieboldiana) bark lectin and sialyloligosaccharides as detected by 1H-NMR.
The interaction of Japanese elderberry bark lectin (Sambucus sieboldiana agglutinin, SSA) with carbohydrate was investigated by 1H-NMR. When a low affinity ligand, methyl beta-D-galactoside (beta MeGal), was mixed with SSA, each proton signal of beta MeGal was broadened. The signal of H-4 was markedly broad, while those of H-1, OCH3, and H-2 of beta MeGal were rather sharp. The specific broadening of Gal H-4 was more evident when SSA was mixed with methyl-beta-D-lactoside (beta MeLac). Position-dependent signal broadening suggests that beta MeGal binds to SSA such that H-4 is closely involved in the contact region, but H-1, OCH3, and H-2 are far from this region. In the case of a high affinity ligand, Neu5Ac(alpha 2-6)Gal(beta 1-4)Glc(= N6L), ligand signals of the SSA-N6L mixture did not change at all. But when a small amount of N6L was added to the SSA-beta MeGal mixture, the broad signals of bound beta MeGal became dramatically sharp. This indicates that the added N6L molecules liberated the bound beta MeGal from SSA. On the other hand, the sialyllactose with the alpha(2-3)-linkage(= N3L) could not substitute for bound beta MeGal because of its lower affinity. This demonstrates that the competitive binding experiment between two ligands is a useful technique to detect the interaction of lectins with high affinity ligands which could not be observed directly by NMR signal broadening and/or chemical shift change.
Takesada
H
H
Central Research Laboratories, Ajinomoto Co., Inc., Kanagawa.
Shibuya
N
N
Nagashima
N
N
D002240:Carbohydrate Sequence; D005679:G(M3) Ganglioside / Q000031:analogs & derivatives / Q000737:chemistry; D037102:Lectins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008756:Methylgalactosides / Q000737:chemistry; D008759:Methylglycosides / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry*; D037121:Plant Lectins; D054788:Ribosome Inactivating Proteins
D005679:G(M3) Ganglioside; D037102:Lectins; D008756:Methylgalactosides; D008759:Methylglycosides; C075507:N-acetylneuraminosyl-2-6-galactosyl-1-4-glucopyranosyl-1-1-ceramide; D009844:Oligosaccharides; D037121:Plant Lectins; C050600:Sambucus nigra lectins; C016735:sialooligosaccharides; C031552:methyl beta-galactoside; C034207:methyl lactoside; D054788:Ribosome Inactivating Proteins