title: Structural characterization of the pH-denatured states of ferricytochrome-c by synchrotron small angle X-ray scattering.
abstract: The ferricytochrome-c (cyt-c) shows a complex unfolding pathway characterized by a series of stable partially folded states. When titrated with HCl at low ionic strength, two transitions are detected. At pH 2, cyt-c assumes the U1 unfolded state, whereas the successive addition of Cl(-) ion from either HCl or NaCl induces the recompaction to a molten globule conformation (A1 and A2 states, respectively). A second unfolded state (U2) is also observed at pH 12. Recent data evidence different features for the local structure of the heme in the different states. To derive relationships between local and overall conformations, we analyzed the structural characteristics of the different states by synchrotron small angle X-ray scattering. The results show that in the acidic-unfolded U1 form the protein assumes a worm-like conformation, whereas in the alkaline-unfolded U2 state, the cyt-c is globular. Moreover, the molten globule states induced by adding HCl or NaCl to U1 appear structurally different: in the A1 state cyt-c is dimeric and less compact, whereas in the A2 form the protein reverts to a globular-like conformation. According to the local heme structure, a molecular model for the different forms is derived.
authors.affiliation: Dipartimento di Fisica and INFM, Università di Perugia, Via Pascoli, I-06123 Perugia, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Cinelli
authors.forename: F
authors.initials: F
authors.lastname: Spinozzi
authors.forename: R
authors.initials: R
authors.lastname: Itri
authors.forename: S
authors.initials: S
authors.lastname: Finet
authors.forename: F
authors.initials: F
authors.lastname: Carsughi
authors.forename: G
authors.initials: G
authors.lastname: Onori
authors.forename: P
authors.initials: P
authors.lastname: Mariani
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D003574:Cytochrome c Group / Q000737:chemistry*; D019281:Dimerization; D006736:Horses; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009010:Monte Carlo Method; D009206:Myocardium / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D013053:Spectrophotometry; D014965:X-Rays
chemical_list: D003574:Cytochrome c Group