[Molecular structure and immunochemical properties of highly purified hemagglutinin from Clostridium botulinum type A].
A procedure for isolation of highly purified hemagglutinin from a toxic complex of culture filtrates of Cl. botulinum type A is described. This procedure includes precipitation with (NH4)2SO4, chromatography on Sephadex G-100, G-200 and DEAE-cellulose, specific adsorption on human erythrocytes and affinity chromatography. Using polyacrylamide gel electrophoresis, it was shown that hemagglutinin is a heteropolymeric protein consisting of a monomer (Mr 53 000) and a trimer (Mr 160 000). The monomer is made up of two subunits with Mr 13 000 and one subunit with Mr 27 000 covalently linked by SS-crosslinks. The number and nature of the SS-crosslinks and SH-groups in the protein molecule were determined and a hypothetical structural model of hemagglutinin was proposed. Using immunochemical analysis, it was shown that some (but not all) serological properties of the highly purified protein from Cl. botulinum type A and of its partially purified counterpart are similar to those of hemagglutinin from Cl. botulinum type B.
D002846:Chromatography, Affinity / Q000379:methods; D003014:Clostridium botulinum / Q000276:immunology*; D004220:Disulfides / Q000032:analysis; D004912:Erythrocytes / Q000276:immunology; D006388:Hemagglutinins / Q000302:isolation & purification*; D006801:Humans; D005779:Immunodiffusion; D046911:Macromolecular Substances; D008970:Molecular Weight; D011487:Protein Conformation
D004220:Disulfides; D006388:Hemagglutinins; D046911:Macromolecular Substances
Issledovanie molekuliarnoĭ struktury i immunokhimicheskikh svoĭstv vysokoochishchennogo gemaggliutinina Clostridium botulinum tipa A.