title: Separation of ricin A- and B-chains after dithiothreitol reduction.
abstract: After complete cleavage of ricin interchain disulfide bridge by 0.05 M dithiothreitol in nondenaturing conditions at 37 degrees C during 1 h 30 min, A- and B-chains were separated on a lactosaminyl-aminoethyl Biogel P-150 column at 4 degrees C, in the presence of 0.01 M dithiothreitol and 0.5 M MgCl2. A- and B-chains have been characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunology. Their specific activities have been tested by protein synthesis inhibition in a cell-free assay (rabbit reticulocyte lysate) and on whole cells (Zajdela hepatoma cells) and by hemagglutination. From these tests, the apparent cross contamination of the chains was about 0.1%.
authors.affiliation: CNRS ER 64, UER Biomédicale des Saints-Pères, Paris, France.
authors.forename: F
authors.initials: F
authors.lastname: Emmanuel
authors.forename: E
authors.initials: E
authors.lastname: Turpin
authors.forename: A
authors.initials: A
authors.lastname: Alfsen
authors.forename: J P
authors.initials: JP
authors.lastname: Frénoy
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D004229:Dithiothreitol; D004591:Electrophoresis, Polyacrylamide Gel; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D007120:Immunochemistry; D066298:In Vitro Techniques; D008970:Molecular Weight; D010084:Oxidation-Reduction; D014176:Protein Biosynthesis; D012276:Ricin / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D012276:Ricin; D004229:Dithiothreitol