Application of millisecond time-resolved solid state NMR to the kinetics and mechanism of melittin self-assembly.

ABSTRACT: Common experimental approaches for characterizing structural conversion processes such as protein folding and self-assembly do not report on all aspects of the evolution from an initial state to the final state. Here, we demonstrate an approach that is based on rapid mixing, freeze-trapping, and low-temperature solid-state NMR (ssNMR) with signal enhancements from dynamic nuclear polarization (DNP). Experiments on the folding and tetramerization of the 26-residue peptide melittin following a rapid pH jump show that multiple aspects of molecular structure can be followed with millisecond time resolution, including secondary structure at specific isotopically labeled sites, intramolecular and intermolecular contacts between specific pairs of labeled residues, and overall structural order. DN

SUBMITTER: Jeon J 

PROVIDER: S-EPMC6708326 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature