title: Measurement of energy landscape roughness of folded and unfolded proteins.
abstract: The dynamics of protein conformational changes, from protein folding to smaller changes, such as those involved in ligand binding, are governed by the properties of the conformational energy landscape. Different techniques have been used to follow the motion of a protein over this landscape and thus quantify its properties. However, these techniques often are limited to short timescales and low-energy conformations. Here, we describe a general approach that overcomes these limitations. Starting from a nonnative conformation held by an aromatic disulfide bond, we use time-resolved spectroscopy to observe nonequilibrium backbone dynamics over nine orders of magnitude in time, from picoseconds to milliseconds, after photolysis of the disulfide bond. We find that the reencounter probability of residues that initially are in close contact decreases with time following an unusual power law that persists over the full time range and is independent of the primary sequence. Model simulations show that this power law arises from subdiffusional motion, indicating a wide distribution of trapping times in local minima of the energy landscape, and enable us to quantify the roughness of the energy landscape (4-5 k(B)T). Surprisingly, even under denaturing conditions, the energy landscape remains highly rugged with deep traps (>20 k(B)T) that result from multiple nonnative interactions and are sufficient for trapping on the millisecond timescale. Finally, we suggest that the subdiffusional motion of the protein backbone found here may promote rapid folding of proteins with low contact order by enhancing contact formation between nearby residues.
authors.affiliation: Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, United Kingdom.
authors.forename: Lilia
authors.initials: L
authors.lastname: Milanesi
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Waltho
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Hunter
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Shaw
authors.forename: Godfrey S
authors.initials: GS
authors.lastname: Beddard
authors.forename: Gavin D
authors.initials: GD
authors.lastname: Reid
authors.forename: Sagarika
authors.initials: S
authors.lastname: Dev
authors.forename: Martin
authors.initials: M
authors.lastname: Volk
mesh_terms: D008958:Models, Molecular; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins