title: A photochemically induced dynamic nuclear polarization study of denatured states of lysozyme.
abstract: Photochemically induced dynamic nuclear polarization (photo-CIDNP) techniques have been used to examine denatured states of lysozyme produced under a variety of conditions. 1H CIDNP difference spectra of lysozyme denatured thermally, by the addition of 10 M urea, or by the complete reduction of its four disulfide bonds were found to differ substantially not only from the spectrum of the native protein but also from that expected for a completely unstructured polypeptide chain. Specifically, denatured lysozyme showed a much reduced enhancement of tryptophan relative to tyrosine than did a mixture of blocked amino acids with the same composition as the intact protein. By contrast, the CIDNP spectrum of lysozyme denatured in dimethyl sulfoxide solution was found to be similar to that expected for a random coil. It is proposed that nonrandom hydrophobic interactions are present within the denatured states of lysozyme in aqueous solution and that these reduce the reactivity of tryptophan residues relative to tyrosine residues. Characterization of such interactions is likely to be of considerable significance for an understanding of the process of protein folding.
authors.affiliation: Physical Chemistry Laboratory, University of Oxford, U.K.
authors.forename: R W
authors.initials: RW
authors.lastname: Broadhurst
authors.forename: C M
authors.initials: CM
authors.lastname: Dobson
authors.forename: P J
authors.initials: PJ
authors.lastname: Hore
authors.forename: S E
authors.initials: SE
authors.lastname: Radford
authors.forename: M L
authors.initials: ML
authors.lastname: Rees
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D002645:Chickens; D006146:Guanidines / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D009113:Muramidase / Q000737:chemistry*; D010777:Photochemistry; D011489:Protein Denaturation; D014508:Urea / Q000737:chemistry
chemical_list: D000596:Amino Acids; D006146:Guanidines; D014508:Urea; D009113:Muramidase