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

Title
NMR characterization of cooperativity: fast ligand binding coupled to slow protein dimerization
Publication Date
2014
Author(s)
Huma, Zil E
Ludeman, Justin P
Wilkinson, Brendan
( author )
OrcID: https://orcid.org/0000-0003-1866-6540
Email: bwilkin7@une.edu.au
UNE Id une-id:bwilkin7
Payne, Richard J
Stone, Martin J
Type of document
Journal Article
Language
en
Entity Type
Publication
Publisher
Royal Society of Chemistry
Place of publication
United Kingdom
DOI
10.1039/c4sc00131a
UNE publication id
une:20870
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.
Link
Citation
Chemical Science, 5(7), p. 2783-2788
ISSN
2041-6539
2041-6520
Start page
2783
End page
2788

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