Structure determination of high-energy states in a dynamic protein ensemble.

ABSTRACT: Macromolecular function frequently requires that proteins change conformation into high-energy states^1-4. However, methods for solving the structures of these functionally essential, lowly populated states are lacking. Here we develop a method for high-resolution structure determination of minorly populated states by coupling NMR spectroscopy-derived pseudocontact shifts⁵ (PCSs) with Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion⁶ (PCS-CPMG). Our approach additionally defines the corresponding kinetics and thermodynamics of high-energy excursions, thereby characterizing the entire free-energy landscape. Using a large set of simulated data for adenylate kinase (Adk), calmodulin and Src kinase, we find that high-energy PCSs accurately determine high-e

SUBMITTER: Stiller JB 

PROVIDER: S-EPMC9126080 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature