Purification and properties of an anti-B hemagglutinin produced by Streptomyces sp.
An anti-B hemagglutinin was purified to homogeneity from the culture filtrate of a strain of Streptomyces sp. by affinity chromatography. The Streptomyces hemagglutinin was adsorbed to insolubilized gum arabic and eluted with 1 M NaCl containing 1 M D-galactose. The purified hemagglutinin is thought to be homogeneous judging from sodium dodecyl sulfate-polyacrylamide gel electrophoresis at pH 7.2, disc gel electrophoresis at pH 4.3, isoelectric focusing, and ultracentrifugation. The molecular weight was estimated to be 11,000 from results of gel filtration in 6 M guanidine hydrochloride (Gdn-HCl), sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and sedimentation equilibrium analysis. The amino acid analyses revealed that the hemagglutinin contained large amounts of alanine, glycine, and valine, 47% of the total amino acid residues, and no phenylalanine. Carbohydrate analysis demonstrated that the hemagglutinin might not be a glycoprotein. The circular dichroic (CD) spectrum of the protein is quite different from those of usual proteins in having a large positive peak at 226 nm (theta = 10,000) and a negative band at 212 nm (theta =-2600). The hemagglutinin showed a typical precipitation curve with gum arabic, and agglutinated human blood group B erythrocytes 256 times as strongly as A or O erythrocytes. These activities were not affected by pH (from 4 to 12). The anti-B activity was further confirmed by serological tests. The hemagglutination-inhibition studies indicated that D-galactose was inhibitory, but alpha-D-galactosides were not necessarily better inhibitors than beta-D-galactosides. L-Rhamnose was the best inhibitor among the monosaccharides tested, and L-arabinose and D-fucose were also inhibitory.
D000017:ABO Blood-Group System; D000373:Agglutinins / Q000032:analysis*; D000596:Amino Acids / Q000032:analysis; D002942:Circular Dichroism; D006386:Hemagglutination Tests; D006388:Hemagglutinins / Q000032:analysis* / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011134:Polysaccharides; D011485:Protein Binding; D011487:Protein Conformation; D013302:Streptomyces / Q000032:analysis*
D000017:ABO Blood-Group System; D000373:Agglutinins; D000596:Amino Acids; D006388:Hemagglutinins; D011134:Polysaccharides