Immunochemical studies on the N-acetyllactosamine beta-(1----6)-linked trisaccharide specificity of Ricinus communis agglutinin.
The combining site of Ricinus communis agglutinin (RCA1) was studied by quantitative precipitin and precipitin inhibition assays. Of 31 complex carbohydrates tested, all except active and inactive antifreeze glycoproteins, Streptococcus group C polysaccharide, and native rat salivary glycoprotein, reacted strongly, and 22 completely precipitated the lectin, indicating that RCA1 has both a broad range of affinity and a low solubility of its carbohydrate-bound complex. Of the monosaccharides and glycosides tested for inhibition of precipitation, p-nitrophenyl beta-D-galactopyranoside was the best. It was about 6.4 times better than methyl beta-D-galactopyranoside. The beta anomer of glycosides of D-galactose was much more potent than the corresponding alpha anomer. Among the oligosaccharides tested, beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal was the best inhibitor, which was approximately 2/3 as active as p-nitrophenyl beta-D-galactopyranoside. It was approximately 1.4 times as active as beta-D-Gal-(1----4)-D-GlcNAc (N-acetyllactosamine), twice as active as beta-D-Gal-(1----3)-D-GlcNAc, and 4.5 times more active than lacto-N-tetraose. From the results, it can be concluded that; (a) hydrophobic interaction is important for binding; (b) the combining site of this lectin is at least as large as a trisaccharide; and (c) of the compounds studied, the trisaccharide beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal was the most complementary to the human blood group I Ma determinant beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----6)-D-Gal.
Wu
A M
AM
Department of Veterinary Pathology, Texas A & M University, College of Veterinary Medicine, College Station 77843.
Sugii
S
S
Gruezo
F G
FG
Kabat
E A
EA
D000606:Amino Sugars / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D005260:Female; D006736:Horses; D006801:Humans; D007150:Immunohistochemistry; D037102:Lectins / Q000378:metabolism*; D037121:Plant Lectins; D013379:Substrate Specificity; D013552:Swine; D014312:Trisaccharides / Q000378:metabolism*
D000606:Amino Sugars; D037102:Lectins; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014312:Trisaccharides; C000458:N-acetyllactosamine