title: Acid denatured apo-cytochrome c is a random coil: evidence from small-angle X-ray scattering and dynamic light scattering.
abstract: The conformation of a denatured protein has been investigated, since the experimental data on the structure of denatured proteins have been incomplete until now. The Stokes' radius Rs and the radius of gyration Rg of apo-cytochrome c at pH 2.3 have been determined by dynamic light scattering and small-angle X-ray scattering, respectively. The values of these structure parameters, extrapolated to zero protein concentration, are Rs = 3.0 nm and Rg = 4.6 nm. The ratio Rg.Rs-1 is a sensitive indicator of the molecular conformation. The ratio of 1.55 obtained by us is typical for a random-coil polymer. The persistence length--the characteristic of the molecular flexibility--was determined to be a = 1.81 nm. From this results the root-mean-square average end-to-end distance of the molecules [h2] 1/2 = 11.2 nm and the characteristic ratio [h2]/npl2p = 8.43, where np = 104 is the number of amino acid residues and lp the distance between C alpha-atoms. We obtained a second virial coefficient A2 = 8.2.10(-3) mol cm3 g-2. The experimentally determined structure parameters are in approximate agreement with those predicted by Flory and others for an unperturbed, randomly coiled polypeptide. The expansion factor lies between 1.1 and 1.2. In conclusion, we have shown that apo-cytochrome c at pH 2.3 and at low concentrations has the conformation of a perturbed random coil with repulsive potentials between the chain segments.
authors.affiliation: Central Institute of Molecular Biology, Berlin-Buch F.R.G.
authors.forename: G
authors.initials: G
authors.lastname: Damaschun
authors.forename: H
authors.initials: H
authors.lastname: Damaschun
authors.forename: K
authors.initials: K
authors.lastname: Gast
authors.forename: C
authors.initials: C
authors.lastname: Gernat
authors.forename: D
authors.initials: D
authors.lastname: Zirwer
mesh_terms: D000818:Animals; D001056:Apolipoproteins C / Q000737:chemistry* / Q000378:metabolism; D006736:Horses; D006863:Hydrogen-Ion Concentration; D008027:Light; D011489:Protein Denaturation; D012542:Scattering, Radiation; D014961:X-Ray Diffraction
chemical_list: D001056:Apolipoproteins C