Mapping Distinct Sequences of Structure Formation Differentiating Multiple Folding Pathways of a Small Protein

J Am Chem Soc. 2021 Jan 27;143(3):1447-1457. doi: 10.1021/jacs.0c11097. Epub 2021 Jan 12.

Abstract

To determine experimentally how the multiple folding pathways of a protein differ, in the order in which the structural parts are assembled, has been a long-standing challenge. To resolve whether structure formation during folding can progress in multiple ways, the complex folding landscape of monellin has been characterized, structurally and temporally, using the multisite time-resolved FRET methodology. After an initial heterogeneous polypeptide chain collapse, structure formation proceeds on parallel pathways. Kinetic analysis of the population evolution data across various protein segments provides a clear structural distinction between the parallel pathways. The analysis leads to a phenomenological model that describes how and when discrete segments acquire structure independently of each other in different subensembles of protein molecules. When averaged over all molecules, structure formation is seen to progress as α-helix formation, followed by core consolidation, then β-sheet formation, and last end-to-end distance compaction. Parts of the protein that are closer in the primary sequence acquire structure before parts separated by longer sequence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fluorescence Resonance Energy Transfer
  • Kinetics
  • Magnoliopsida / chemistry
  • Plant Proteins / chemistry*
  • Protein Folding*

Substances

  • Plant Proteins
  • monellin protein, Dioscoreophyllum cumminsii