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Experimental Evidence for Millisecond–Timescale Structural Evolution Following the Microsecond–Timescale Folding of a Small Protein

C. Blake Wilson , Wai-Ming Yau, and Robert Tycko ^*

Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0520, USA

^*Corresponding author: robertty@mail.nih.gov

Phys. Rev. Lett. 132, 048402 – Published 24 January, 2024

DOI: https://doi.org/10.1103/PhysRevLett.132.048402

Abstract

Prior work has shown that small proteins can fold (i.e., convert from unstructured to structured states) within $10 μ s$ . Here we use time-resolved solid state nuclear magnetic resonance (ssNMR) methods to show that full folding of the 35-residue villin headpiece subdomain (HP35) requires a slow annealing process that has not been previously detected. ${}^{13}C$ ssNMR spectra of frozen HP35 solutions, acquired with a variable time $τ_{e}$ at $30 ° C$ after rapid cooling from $95 ° C$ and before rapid freezing, show changes on the 3–10 ms timescale, attributable to slow rearrangements of protein sidechains during $τ_{e}$ .

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Protein Folding Can Be Surprisingly Slow

Published 24 January, 2024

Researchers have used nuclear magnetic resonance to observe a previously unseen intermediate state in which the protein lingers for an unexpectedly long time.

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