Characterizing Protein-Protein Interactions Using Solution NMR Spectroscopy

Jose Luis Ortega-Roldan; Martin Blackledge; Malene Ringkjøbing Jensen

doi:10.1007/978-1-4939-7759-8_5

Characterizing Protein-Protein Interactions Using Solution NMR Spectroscopy

Methods Mol Biol. 2018:1764:73-85. doi: 10.1007/978-1-4939-7759-8_5.

Authors

Jose Luis Ortega-Roldan¹, Martin Blackledge², Malene Ringkjøbing Jensen³

Affiliations

¹ School of Biosciences, University of Kent, Canterbury, UK.
² Univ. Grenoble Alpes, CEA, CNRS, IBS, Grenoble, France.
³ Univ. Grenoble Alpes, CEA, CNRS, IBS, Grenoble, France. malene.ringkjobing-jensen@ibs.fr.

PMID: 29605909
DOI: 10.1007/978-1-4939-7759-8_5

Abstract

In this chapter, we describe how NMR chemical shift titrations can be used to study the interaction between two proteins with emphasis on mapping the interface of the complex and determining the binding affinity from a quantitative analysis of the experimental data. In particular, we discuss the appearance of NMR spectra in different chemical exchange regimes (fast, intermediate, and slow) and how these regimes affect NMR data analysis.

Keywords: Binding affinity; Chemical exchange; Chemical shift titration; Dissociation constant; Protein-protein interactions; Solution NMR spectroscopy.

MeSH terms

Binding Sites
Humans
Kinetics
Magnetic Resonance Spectroscopy / methods*
Models, Molecular
Protein Binding
Ubiquitin / chemistry
Ubiquitin / metabolism*
src Homology Domains*

Substances

Ubiquitin