title: Characterization of the stable, acid-induced, molten globule-like state of staphylococcal nuclease.
abstract: Titration of a salt-free solution of native staphylococcal nuclease by HCl leads to an unfolding transition in the vicinity of pH 4, as determined by near- and far-UV circular dichroism. At pH 2-3, the protein is substantially unfolded. The addition of further HCl results in a second transition, this one to a more structured species (the A state) with the properties of an expanded molten globule, namely substantial secondary structure, little or no tertiary structure, relatively compact size as determined by hydrodynamic radius, and the ability to bind the hydrophobic dye 1-anilino-8-naphthalene sulfonic acid. The addition of anions, in the form of neutral salts, to the acid-unfolded state at pH 2 also causes a transition leading to the A state. Fourier transform infrared analysis of the amide I band was used to compare the amount and type of secondary structure in the native and A states. A significant decrease in alpha-helix structure, with a corresponding increase in beta or extended structure, was observed in the A state, compared to the native state. A model to account for such compact denatured states is proposed.
authors.affiliation: Department of Chemistry, University of California, Santa Cruz 95064.
authors.forename: A L
authors.initials: AL
authors.lastname: Fink
authors.forename: L J
authors.initials: LJ
authors.lastname: Calciano
authors.forename: Y
authors.initials: Y
authors.lastname: Goto
authors.forename: M
authors.initials: M
authors.lastname: Nishimura
authors.forename: S A
authors.initials: SA
authors.lastname: Swedberg
mesh_terms: D000817:Anilino Naphthalenesulfonates; D000838:Anions; D002942:Circular Dichroism; D004795:Enzyme Stability; D006851:Hydrochloric Acid; D006863:Hydrogen-Ion Concentration; D008836:Micrococcal Nuclease / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013055:Spectrophotometry, Infrared
chemical_list: D000817:Anilino Naphthalenesulfonates; D000838:Anions; C027132:1-anilino-8-naphthalenesulfonate; D008836:Micrococcal Nuclease; D006851:Hydrochloric Acid