Immunochemical studies on blood groups. Structures and immunochemical properties of nine oligosaccharides from B-active and non-B-active blood group substances of horse gastric mucosae.

Oligosaccharides from base-borohydride-treated B-active and non-B-active glycoproteins of horse stomach mucosae were purified chromatographically on Bio-Gel P-2, charcoal-Celite, paper and high pressure liquid chromatography. From colorimetric and gas-liquid Chromatographic analyses, methylation, quantitative periodate oxidation and Smith degradation, structures of nine Oligosaccharides are proposed. Seven have not been previously described. The oligosaccharide isolated in largest amount in the B-active reduced tetrasaccharide analogous to an A-active reduced oligosaccharide from pig submaxillary mucin, and a reduced octasaccharide, the largest isolated, has two B determinants and may represent full expression of B-specific biosynthetic potential of the mucosal lining. Three B-active and one non-B-active oligosaccharide possessed the core structure previously identified in Oligosaccharides from human blood group, B, HLe^b, Le^a and precursor I substances. Two non-B-active and one B-active compound inhibited the cross reaction of type XIV horse antipneumococcal sera with blood group substances. Terminal nonreducing α-linked dGlcNAc (d-2-acetamido-2-deoxyglucopyranose), previously found in Oligosaccharides of hog blood group substances, was also present in a tetrassarcharide of the non-B-active material. Oligosaccharides released from blood group glycoproteins of horse stomach mucosae are smaller and hence less heterogeneous than those from human ovarian cyst and perhaps hog A + H and human gastric mucosae.     

Immunochemical studies on blood groups: Purification,chemical and immunochemical properties of blood group-active glycoproteins from horse gastric mucosae

Digestion of the gastric mucosae of 10 horses with pepsin or Pronase was followed by phenol/ethanol fractionation. Chemical and immunochemical examination of the fractions showed the mucosae to possess various combinations of A, B and H activities. Most were B-active, three had weak A activity, one had strong H activity and the remainder were weakly H-active; one mucosa possessed neither A, B nor H activity. Digestion with pepsin or Pronase of different portions of the same mucosa yielded products equivalent in serological and most chemical properties. Materials digested by Pronase tended to have less peptide nitrogen than those treated with pepsin. Fractions with the strongest serological activities contained significantly higher amounts of carbohydrate and lesser amounts of peptide nitrogen than those with weak A, B or H activity or with no activity. All mucosae, independent of their A, B or H activity, reacted with concanavalin A. The fractions precipitable by 10% ethanol from 90% phenol reacted most strongly.     

Immunochemical studies on bacterial blood group substances. VIII. The structure of oligosaccharides from blood group H-active polysaccharide of Escherichia coli 2B-V.

Blood group H-active polysaccharide has been prepared from "smooth" strain Escherichia coli 2B-V by Freeman's method, alpha-Fucosidase derived from Bacillus fluminans caused the liberation of fucose from this polysaccharide, together with concomitant loss of blood group H activity. The results of quantitative microanalysis, borohydride reduction, the Morgan-Elson reaction and enzymic hydrolysis with betagalactosidase using isolated oligosaccharides obtained by partial acid hydrolysis indicated that the O-specific side chain of the polysaccharide has a pentassaccharide unit which is beta-D-Gal-(1 leads to 3)-D-GalNAc-(1 leads to 3)-D-GalNAc-Fuc with a D-glucose residue bound at some undetermined point on this structure. It was considered that terminal non-reducing fucose of the polysaccharide was liberated by partial acid hydrolysis.     

Immunochemical studies on blood groups. 53. A study of various conditions of alkaline borohydride degradation on human and hog blood group substances and on known oligosaccharides

Blood group A, B, H, Le^a, Le^b, and I substances, their products of periodate oxidation and Smith degradation, and disaccharides containing 3-O-substituted reducing N-acetylhexosamines were treated with base-borohydride under three defined sets of conditions. Procedures for the assay and quantitation of the possible reduced base-degradation products, including hexenetetrol(s), 3-deoxygalactitol, galactitol, reduced chromogens, N-acetylglucosaminitol, and N-acetylgalactosaminitol are described. Extensive degradation occurred by two methods. 1 m NaBH₄ in 0.05 n NaOH at 50 ° cleaves the glycosidic linkage of the oligosaccharide chains from serine and threonine with reduction of the terminal-reducing N-acetylgalactosamine with minimal base degradation. The method is useful for isolation of complete reduced oligosaccharides from blood group substances; the structural implications of the free and oligosaccharide-bound N-acetylgalactosaminitol released are discussed.