title: Catalysis of protein folding by agarose-immobilized protein disulfide isomerase.
abstract: Protein disulfide isomerase (PDI) catalyzes the formation and rearrangement of disulfide bonds during protein folding. PDI coupled to cyanogen bromide-activated agarose retains its catalytic activity, and a column of this material increases both the rate and the yield for folding disulfide-containing proteins. For reduced, denatured ribonuclease, the overall yield of fully active ribonuclease isolated from the PDI column in one pass was 85-98% of the applied protein. Under the same conditions in the absence of PDI, ribonuclease regained only 16% of its native activity. The oxidative folding of reduced denatured lysozyme is complicated by aggregation so that in the absence of PDI optimal yields of only < or = 25% are obtained at lysozyme concentrations of 1.6 mg/ml. When reduced, denatured lysozyme (1.6 mg/ml) is passed over a PDI column in 1-2 M urea in the presence of a glutathione redox buffer, the specific activity of the recovered lysozyme is identical to that of the native enzyme and the total recovery of the applied protein is 50-65%.
authors.affiliation: Verna and Marrs McLean Department of Biochemistry, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: N A
authors.initials: NA
authors.lastname: Morjana
authors.forename: H F
authors.initials: HF
authors.lastname: Gilbert
mesh_terms: D000818:Animals; D002384:Catalysis; D002417:Cattle; D002645:Chickens; D002853:Chromatography, Liquid; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D007535:Isomerases / Q000378:metabolism*; D009113:Muramidase / Q000378:metabolism; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D019704:Protein Disulfide-Isomerases; D017510:Protein Folding; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D012685:Sepharose; D012995:Solubility
chemical_list: D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D012685:Sepharose; D012259:Ribonuclease, Pancreatic; D009113:Muramidase; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; D005978:Glutathione