title: Electrostatic optimization of the conformational energy landscape in a metamorphic protein.
abstract: The equilibrium unfolding reaction of Ltn, a metamorphic C-class chemokine, was monitored by tryptophan fluorescence to determine unfolding free energies. Measurements revealed that addition of 150 mM NaCl stabilized the Ltn chemokine fold by approximately 1 kcal/mol. Specific mutations involving Arg23 and Arg43 also increased the stability by 1 kcal/mol, suggesting their involvement in chloride ion coordination. This interaction was confirmed by nuclear magnetic resonance (NMR) salt titration studies that revealed chemical shift perturbations localized to these residues and backbone amides within the proximal 40s loop. The effects of NaCl on the free energy landscape were further verified by ZZ-exchange NMR spectroscopy. Our results suggest that changes in the electrostatic environment modulate the Gibbs free energy of folding and alter the forward and reverse rates of interconversion. These results demonstrate how solution ions can promote metamorphic folding by adjusting the relative stabilities of two unrelated Ltn native-state structures.
authors.affiliation: Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, United States.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Tyler
authors.forename: Jamie C
authors.initials: JC
authors.lastname: Wieting
authors.forename: Francis C
authors.initials: FC
authors.lastname: Peterson
authors.forename: Brian F
authors.initials: BF
authors.lastname: Volkman
mesh_terms: D001665:Binding Sites; D019744:Chemokines, C / Q000737:chemistry* / Q000235:genetics; D002712:Chlorides / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011489:Protein Denaturation; D055550:Protein Stability; D012965:Sodium Chloride / Q000494:pharmacology; D055672:Static Electricity; D013816:Thermodynamics; D014364:Tryptophan / Q000737:chemistry
chemical_list: D019744:Chemokines, C; D002712:Chlorides; C092390:XCL1 protein, human; D012965:Sodium Chloride; D014364:Tryptophan