Discovering and Manipulating Protein Conformational Heterogeneity and Function

Proteins must transition between multiple conformations to accomplish their functions in the cell, yet the dominant three-dimensional perspective involves a static crystal structure representing the most populated single conformation. Recent advances have revealed previously "hidden" alternative conformations with the potential to bridge dynamic descriptions from NMR spectroscopy with the static descriptions from X-ray crystallography. However, it remains challenging to determine which of these alternative conformations are collectively responsive to perturbations. As a result, the fundamental interplay between conformational heterogeneity and biological functions such as catalysis and allostery remains poorly understood. To overcome these obstacles, we develop new algorithms to identify hidden alternative conformations and test whether they form cooperative interaction networks across a set of benchmark proteins. We also combine these computational tools with novel X-ray crystallography experiments to characterize the inherent conformational heterogeneity of the diabetes therapeutic target PTP1B and identify new susceptible allosteric sites. Our approach will open new avenues for the design of novel therapeutics and dynamically enabled protein functions.View Large Image | View Hi-Res Image | Download PowerPoint Slide