NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization

Zil E Huma, Justin Peter Ludeman, Brendan Wilkinson, Richard James Payne, Martin Jeremy Stone

Research output: Contribution to journalArticleResearchpeer-review

Abstract

We describe a general approach for analysis of 2D NMR spectra to evaluate the cooperativity of ligand binding and protein dimerization in coupled systems. The approach is applicable to systems in which NMR spectra display separate resonances for monomeric and dimeric species but each resonance shifts in response to ligand binding. Three experimental parameters (monomer chemical shift, dimer chemical shift and relative monomer-dimer peak intensity) are fitted globally, as a function of ligand concentration, to yield equilibrium constants for dimerization, monomer-ligand binding and dimer-ligand binding as well as the cooperativity between ligand binding and dimerization. We have applied the approach to characterise a system in which dimerization of the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2) is coupled to binding of peptides derived from the chemokine receptor CCR2. The global fitting approach allowed evaluation of cooperativity with higher precision than is possible by alternative methods. ? 2014 the Partner Organisations.
Original languageEnglish
Pages (from-to)2783 - 2788
Number of pages6
JournalChemical Science
Volume5
Issue number7
DOIs
Publication statusPublished - 2014

Cite this

Huma, Zil E ; Ludeman, Justin Peter ; Wilkinson, Brendan ; Payne, Richard James ; Stone, Martin Jeremy. / NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization. In: Chemical Science. 2014 ; Vol. 5, No. 7. pp. 2783 - 2788.
@article{55ba42b17f2d4b649bee0b33233a6382,
title = "NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization",
abstract = "We describe a general approach for analysis of 2D NMR spectra to evaluate the cooperativity of ligand binding and protein dimerization in coupled systems. The approach is applicable to systems in which NMR spectra display separate resonances for monomeric and dimeric species but each resonance shifts in response to ligand binding. Three experimental parameters (monomer chemical shift, dimer chemical shift and relative monomer-dimer peak intensity) are fitted globally, as a function of ligand concentration, to yield equilibrium constants for dimerization, monomer-ligand binding and dimer-ligand binding as well as the cooperativity between ligand binding and dimerization. We have applied the approach to characterise a system in which dimerization of the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2) is coupled to binding of peptides derived from the chemokine receptor CCR2. The global fitting approach allowed evaluation of cooperativity with higher precision than is possible by alternative methods. ? 2014 the Partner Organisations.",
author = "Huma, {Zil E} and Ludeman, {Justin Peter} and Brendan Wilkinson and Payne, {Richard James} and Stone, {Martin Jeremy}",
year = "2014",
doi = "10.1039/c4sc00131a",
language = "English",
volume = "5",
pages = "2783 -- 2788",
journal = "Chemical Science",
issn = "2041-6520",
publisher = "The Royal Society of Chemistry",
number = "7",

}

NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization. / Huma, Zil E; Ludeman, Justin Peter; Wilkinson, Brendan; Payne, Richard James; Stone, Martin Jeremy.

In: Chemical Science, Vol. 5, No. 7, 2014, p. 2783 - 2788.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization

AU - Huma, Zil E

AU - Ludeman, Justin Peter

AU - Wilkinson, Brendan

AU - Payne, Richard James

AU - Stone, Martin Jeremy

PY - 2014

Y1 - 2014

N2 - We describe a general approach for analysis of 2D NMR spectra to evaluate the cooperativity of ligand binding and protein dimerization in coupled systems. The approach is applicable to systems in which NMR spectra display separate resonances for monomeric and dimeric species but each resonance shifts in response to ligand binding. Three experimental parameters (monomer chemical shift, dimer chemical shift and relative monomer-dimer peak intensity) are fitted globally, as a function of ligand concentration, to yield equilibrium constants for dimerization, monomer-ligand binding and dimer-ligand binding as well as the cooperativity between ligand binding and dimerization. We have applied the approach to characterise a system in which dimerization of the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2) is coupled to binding of peptides derived from the chemokine receptor CCR2. The global fitting approach allowed evaluation of cooperativity with higher precision than is possible by alternative methods. ? 2014 the Partner Organisations.

AB - We describe a general approach for analysis of 2D NMR spectra to evaluate the cooperativity of ligand binding and protein dimerization in coupled systems. The approach is applicable to systems in which NMR spectra display separate resonances for monomeric and dimeric species but each resonance shifts in response to ligand binding. Three experimental parameters (monomer chemical shift, dimer chemical shift and relative monomer-dimer peak intensity) are fitted globally, as a function of ligand concentration, to yield equilibrium constants for dimerization, monomer-ligand binding and dimer-ligand binding as well as the cooperativity between ligand binding and dimerization. We have applied the approach to characterise a system in which dimerization of the chemokine monocyte chemoattractant protein-1 (MCP-1/CCL2) is coupled to binding of peptides derived from the chemokine receptor CCR2. The global fitting approach allowed evaluation of cooperativity with higher precision than is possible by alternative methods. ? 2014 the Partner Organisations.

UR - http://pubs.rsc.org.ezproxy.lib.monash.edu.au/en/content/articlepdf/2014/sc/c4sc00131a

U2 - 10.1039/c4sc00131a

DO - 10.1039/c4sc00131a

M3 - Article

VL - 5

SP - 2783

EP - 2788

JO - Chemical Science

JF - Chemical Science

SN - 2041-6520

IS - 7

ER -