The inverted chevron plot measured by NMR relaxation reveals a native-like unfolding intermediate in acyl-CoA binding protein.

Proceedings of the National Academy of Sciences of the United States of America
Kaare TeilumMikael Akke

Abstract

The folding kinetics of bovine acyl-CoA binding protein was studied by 15N relaxation dispersion measurements under equilibrium conditions. Relaxation dispersion profiles were measured at several concentrations of guanidine hydrochloride (GuHCl). The unfolding rate constant (k(u)) was determined under conditions favoring folding, for which the folding rate constant (k(f)) dominates the relaxation in stopped-flow kinetic measurements. Conversely, k(f) was determined under conditions favoring unfolding, for which k(u) dominates stopped-flow data. The rates determined by NMR therefore complement those from stopped-flow kinetics and define an "inverted chevron" plot. The combination of NMR relaxation and stopped-flow kinetic measurements allowed determination of k(f) and k(u) in the range from 0.48 M GuHCl to 1.28 M GuHCl. Individually, the stopped-flow and NMR data fit two-state models for folding. However, although the values of k(f) determined by the two methods agree, the values of k(u) do not. As a result, a combined analysis of all data does not comply with a two-state model but indicates that an unfolding intermediate exists on the native side of the dominant energy barrier. The denaturant and temperature dependencies of the...Continue Reading

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Citations

Oct 14, 2008·Journal of Biomolecular NMR·Kristofer Modig, Flemming M Poulsen
Jul 4, 2012·Journal of the American Chemical Society·Vincent A VoelzVijay S Pande
Sep 26, 2012·Journal of the American Chemical Society·Pétur O HeidarssonCiro Cecconi
Dec 23, 2006·Proceedings of the National Academy of Sciences of the United States of America·Armin HoffmannBenjamin Schuler
Jul 14, 2010·Proceedings of the National Academy of Sciences of the United States of America·Susanne W BruunFlemming M Poulsen
Feb 6, 2016·Journal of Biomolecular NMR·Troels E Linnet, Kaare Teilum
May 15, 2007·Biophysical Chemistry·Stefano GianniCarlo Travaglini-Allocatelli
Nov 12, 2013·Journal of Molecular Biology·Valéry OzenneJens Danielsson
Jan 24, 2007·Current Opinion in Structural Biology·David J Brockwell, Sheena E Radford
Apr 1, 2015·Physical Chemistry Chemical Physics : PCCP·Sneha Munshi, Athi N Naganathan
Aug 2, 2020·Analytical Biochemistry·Louise HamborgKaare Teilum
Oct 21, 2010·Journal of the American Chemical Society·Anne DhulesiaChristopher M Dobson

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