title: An evidence for the equilibrium unfolding intermediates of ribonuclease A by tritium labeling method.
abstract: A formation of a molten globule in the unfolding of ribonuclease A could be considered as an evidence supporting a hypothesis on the existence of such intermediates on the pathway of a protein folding. Using a novel technique (tritium labeling method) we have showed that the ribonuclease A equilibrium unfolding in urea and guanidinium chloride (GuCl) solutions proceeds through a formation of intermediates whose properties (compactness, retention of the larger part hydrophobic core, secondary structure, and native-like folding pattern) correspond to the fundamental characteristics of the molten globule state. The both intermediates are the "wet" molten globules (the globule interior contains the water molecules). The results reveal the noticeable distinctions in intermediates structure, first of all, in the extent of their compactness. The urea intermediate is less compact than that in GuCl. It is shown that the refolding of the protein denatured by GuCl results in the formation of the intermediate which enzyme activity is virtually the same as the activity of the native protein.
authors.affiliation: NN Semenov Institute of Chemical Physics, Russian Academy of Sciences, Kosygin St 4, Moscow, Russia. avolynskaya@mail.ru
authors.forename: Aleftina V
authors.initials: AV
authors.lastname: Volynskaya
authors.forename: Eldar A
authors.initials: EA
authors.lastname: Kasumov
authors.forename: Vitalii I
authors.initials: VI
authors.lastname: Goldanskii
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D019791:Guanidine; D057927:Hydrophobic and Hydrophilic Interactions; D066298:In Vitro Techniques; D018021:Lithium Chloride; D010179:Pancreas / Q000201:enzymology; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D014316:Tritium; D014508:Urea
chemical_list: D000596:Amino Acids; D014316:Tritium; D014508:Urea; D012259:Ribonuclease, Pancreatic; D018021:Lithium Chloride; D019791:Guanidine