Recovery of Protein Folding Funnels from Single-Molecule Time Series by Delay Embeddings and Manifold Learning.

The stability and folding of proteins is governed by the underlying single-molecule free energy surface (smFES) mapping the free energy of the molecule as a function of configurational state. Ascertaining the smFES is of great value in understanding and engineering protein structure and function. By integrating tools from dynamical systems theory and nonlinear manifold learning, we describe an approach to reconstruct the multidimensional smFES for a protein from a time series in a single experimentally measurable observable. We employ Takens' delay embeddings to project the time series into a high-dimensional space in which the projected dynamics are C-equivalent to the true system dynamics and employ diffusion maps to recover a low-dimensional reconstruction of the smFES that is equivalent to the true smFES up to a smooth and invertible transformation. We validate the approach in molecular dynamics simulations of Trp-cage, Villin, and BBA to demonstrate that landscapes recovered from univariate time series in the head-to-tail distance are topologically identical-they precisely preserve the metastable states and folding pathways-and topographically approximate-the free energy barrier heights and well depths are approximately preserved-to the true landscapes determined from complete knowledge of all atomic coordinates. We go on to show that the reconstructed landscapes reliably predict temperature denaturation and identify point mutations and groups of mutations critical to folding. These results demonstrate that protein folding funnels can be reconstructed from experimentally measurable time series and used to understand and engineer folding.