title: Experiments and simulations show how long-range contacts can form in expanded unfolded proteins with negligible secondary structure.
abstract: The sizes of unfolded proteins under highly denaturing conditions scale as N(0.59) with chain length. This suggests that denaturing conditions mimic good solvents, whereby the preference for favorable chain-solvent interactions causes intrachain interactions to be repulsive, on average. Beyond this generic inference, the broader implications of N(0.59) scaling for quantitative descriptions of denatured state ensembles (DSEs) remain unresolved. Of particular interest is the degree to which N(0.59) scaling can simultaneously accommodate intrachain attractions and detectable long-range contacts. Here we present data showing that the DSE of the N-terminal domain of the L9 (NTL9) ribosomal protein in 8.3 M urea lacks detectable secondary structure and forms expanded conformations in accord with the expected N(0.59) scaling behavior. Paramagnetic relaxation enhancements, however, indicate the presence of detectable long-range contacts in the denatured-state ensemble of NTL9. To explain these observations we used atomistic thermal unfolding simulations to identify ensembles whose properties are consistent with all of the experimental observations, thus serving as useful proxies for the DSE of NTL9 in 8.3 M urea. Analysis of these ensembles shows that residual attractions are present under mimics of good solvent conditions, and for NTL9 they result from low-likelihood, medium/long-range contacts between hydrophobic residues. Our analysis provides a quantitative framework for the simultaneous observation of N(0.59) scaling and low-likelihood long-range contacts for the DSE of NTL9. We propose that such low-likelihood intramolecular hydrophobic clusters might be a generic feature of DSEs that play a gatekeeping role to protect against aggregation during protein folding.
authors: Department of Chemistry, State University of New York at Stony Brook, Stony Brook, NY 11794-3400, USA.
authors: Wenli
authors: W
authors: Meng
authors: Nicholas
authors: N
authors: Lyle
authors: Bowu
authors: B
authors: Luan
authors: Daniel P
authors: DP
authors: Raleigh
authors: Rohit V
authors: RV
authors: Pappu
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D055592:Biophysical Phenomena; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D058767:Protein Unfolding; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics; D053838:Scattering, Small Angle; D017386:Sequence Homology, Amino Acid; D014508:Urea; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D012269:Ribosomal Proteins; C049650:ribosomal protein L9; D014508:Urea