title: Acceleration of oxidative folding of bovine pancreatic ribonuclease A by anion-induced stabilization and formation of structured native-like intermediates.
abstract: Phosphate anions accelerate the oxidative folding of reduced bovine pancreatic ribonuclease A with dithiothreitol at several temperatures and ionic strengths. The addition of 400 mM phosphate at pH 8.1 increased the regeneration rate of native protein 2.5-fold at 15 degrees C, 3.5-fold at 25 degrees C, and 20-fold at 37 degrees C, compared to the rate in the absence of phosphate. In addition, the effects of other ions on the oxidative folding of RNase A were examined. Fluoride was found to accelerate the formation of native protein under the same oxidizing conditions. In contrast, cations of high charge density or ions with low charge density appear to have an opposite effect on the folding of RNase A. The catalysis of oxidative folding results largely from an anion-dependent stabilization and formation of tertiary structure in productive disulfide intermediates (des-species). Phosphate and fluoride also accelerate the initial equilibration of unstructured disulfide ensembles, presumably due to non-specific electrostatic and hydrogen bonding effects on the protein and solvent.
authors.affiliation: Baker Laboratory of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853-1301, USA.
authors.forename: L K
authors.initials: LK
authors.lastname: Low
authors.forename: H C
authors.initials: HC
authors.lastname: Shin
authors.forename: M
authors.initials: M
authors.lastname: Narayan
authors.forename: W J
authors.initials: WJ
authors.lastname: Wedemeyer
authors.forename: H A
authors.initials: HA
authors.lastname: Scheraga
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004229:Dithiothreitol / Q000737:chemistry; D004795:Enzyme Stability; D006361:Heating; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010179:Pancreas / Q000737:chemistry*; D010710:Phosphates / Q000737:chemistry; D017510:Protein Folding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D012492:Salts
chemical_list: D010710:Phosphates; D012492:Salts; D012259:Ribonuclease, Pancreatic; D004229:Dithiothreitol