Resolution of thiol-containing proteins by sequential-elution covalent chromatography.
Elution of complex protein mixtures on a matrix containing reactive disulphide bonds (Thiopropyl-Sepharose 6B, Pharmacia) results in immobilisation of thiol-containing molecules. Specific protein fractions can be displaced from the gel using different low-molecular-weight reducing agents. Thus a single sequential elution can separate and resolve thiol-containing proteins in a rapid and convenient step. The method is illustrated with reference to beef liver thiol : disulphide oxidoreductases.
D000818:Animals; D002021:Buffers; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange; D003545:Cysteine; D004220:Disulfides / Q000302:isolation & purification; D004229:Dithiothreitol; D005978:Glutathione; D007535:Isomerases / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology*; D010088:Oxidoreductases / Q000302:isolation & purification*; D011490:Protein Disulfide Reductase (Glutathione) / Q000302:isolation & purification*; D019704:Protein Disulfide-Isomerases
D002021:Buffers; D004220:Disulfides; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione); D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D005978:Glutathione; D003545:Cysteine; D004229:Dithiothreitol