Single-Molecule Dynamics Reveals Cooperative Binding-Folding in Protein Recognition

Open Access
Publisher Website
Google Scholar
Abstract
The study of associations between two biomolecules is the key to understanding molecular function and recognition. Molecular function is often thought to be determined by underlying structures. Here, combining a single-molecule study of protein binding with an energy-landscape–inspired microscopic model, we found strong evidence that biomolecular recognition is determined by flexibilities in addition to structures. Our model is based on coarse-grained molecular dynamics on the residue level with the energy function biased toward the native binding structure (the Go model). With our model, the underlying free-energy landscape of the binding can be explored. There are two distinct conformational states at the free-energy minimum, one with partial folding of CBD itself and significant interface binding of CBD to Cdc42, and the other with native folding of CBD itself and native interface binding of CBD to Cdc42. This shows that the binding process proceeds with a significant interface binding of CBD with Cdc42 first, without a complete folding of CBD itself, and that binding and folding are then coupled to reach the native binding state. The single-molecule experimental finding of dynamic fluctuations among the loosely and closely bound conformational states can be identified with the theoretical, calculated free-energy minimum and explained quantitatively in the model as a result of binding associated with large conformational changes. The theoretical predictions identified certain key residues for binding that were consistent with mutational experiments. The combined study identified fundamental mechanisms and provided insights about designing and further exploring biomolecular recognition with large conformational changes. Biomolecular function (e.g., binding) is often thought to be determined by the underlying molecular structure. There are more and more findings that molecular binding sometimes involves large conformational changes in various stages of cell function. Addressing this issue will answer the critical questions about how molecular function is determined by conformational flexibility and dynamics in addition to structure. Combining a single-molecule fluorescence study of flexible protein binding with an energy-landscape–inspired microscopic molecular dynamics model, the authors found strong evidence that biomolecular recognition is determined by flexibility and large conformational changes in addition to structure. The single-molecule study shows conformational fluctuations of the protein complex that involve bound and loosely bound states, which can be quantitatively explained in the authors' model as a result of cooperative binding. Theoretical predictions about the key residues are consistent with mutational experiments. Identifying the key residues for binding provides a structural basis for designing drugs that will target those critical residues.

This publication has 47 references indexed in Scilit: