Multi-probe Relaxation Dispersion Measurements Increase Sensitivity to Protein Dynamics

Overview

Journal Phys Chem Chem Phys

Publisher Royal Society of Chemistry

Specialties Biophysics
Chemistry

Date 2015 Oct 2

PMID 26426424

Citations 7

Authors

R Bryn Fenwick

David Oyen

Peter E Wright

Affiliations

Soon will be listed here.

Abstract

Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion measurements are a valuable tool for the characterization of structural transitions on the micro-millisecond timescale. While the measurement of (15)N relaxation dispersion is now routine, the measurements with alternative nuclei remain limited. Here we report (15)N as well as (1)H R2 relaxation dispersion measurements of the N23PP/S148A "dynamic knockout" mutant of dihydrofolate reductase. The (1)H dispersion measurements are complementary to (15)N data as many additional residues are observed to have dispersive behavior for the (1)H nucleus. Simultaneous fitting of the dispersion profiles for the two nuclei increases the accuracy of exchange parameters determined for individual residues and clustered groups of residues. The different sensitivity of the two nuclei to changes in backbone torsional angles, ring currents, and hydrogen bonding effects provides important insights into the nature of the structural changes that take place during the exchange process. We observe clear evidence of direct and indirect hydrogen bond effects for the (15)N and (1)H chemical shift changes in the active-site, modulation of ring current shielding in the CD-loop and backbone torsional changes in a cluster of residues associated with the C-terminus. This work demonstrates the power of combined (1)H and (15)N probes for the study of backbone dynamics on the micro-millisecond timescale though the analysis of chemical shift changes.

Citing Articles

Conformational Dynamics of an Amyloidogenic Intermediate of Transthyretin: Implications for Structural Remodeling and Amyloid Formation.

Leach B, Ferguson J, Morgan G, Sun X, Kroon G, Oyen D J Mol Biol. 2024; 436(16):168673.

PMID: 38909653 PMC: 11410348. DOI: 10.1016/j.jmb.2024.168673.

Role of Active Site Loop Dynamics in Mediating Ligand Release from Dihydrofolate Reductase.

Singh A, Fenwick R, Dyson H, Wright P Biochemistry. 2021; 60(35):2663-2671.

PMID: 34428034 PMC: 8879413. DOI: 10.1021/acs.biochem.1c00461.

Modeling of Hidden Structures Using Sparse Chemical Shift Data from NMR Relaxation Dispersion.

Fenwick R, Oyen D, van den Bedem H, Dyson H, Wright P Biophys J. 2020; 120(2):296-305.

PMID: 33301748 PMC: 7840400. DOI: 10.1016/j.bpj.2020.11.2267.

General Expressions for Carr-Purcell-Meiboom-Gill Relaxation Dispersion for N-Site Chemical Exchange.

Koss H, Rance M, Palmer 3rd A Biochemistry. 2018; 57(31):4753-4763.

PMID: 30040382 PMC: 6315113. DOI: 10.1021/acs.biochem.8b00370.

Slow Dynamics of Tryptophan-Water Networks in Proteins.

Fenwick R, Oyen D, Dyson H, Wright P J Am Chem Soc. 2017; 140(2):675-682.

PMID: 29256600 PMC: 5771866. DOI: 10.1021/jacs.7b09974.

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