Solution NMR Spectroscopy for Characterizing Protein–Glycosaminoglycan Interactions

Prem Raj B. Joseph, Krishna Mohan Sepuru, Krishna Mohan Poluri, Krishna Rajarathnam

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Solution nuclear magnetic resonance (NMR) spectroscopy and, in particular, chemical shift perturbation (CSP) titration experiments are ideally suited for mapping and characterizing the binding interface of macromolecular complexes.1H-15N-HSQC-based CSP studies have become the method of choice due to their simplicity, short-time requirements, and minimal working knowledge of NMR. CSP studies for characterizing protein–glycosaminoglycan (GAG) interactions can be challenging due to binding-induced aggregation/precipitation and/or poor quality data. In this chapter, we discuss how optimizing experimental conditions such as protein concentration, choice of buffer pH, ionic strength, and GAG size, as well as sensitivity of NMR instrumentation can overcome these roadblocks to obtain meaningful structural insights into protein–GAG interactions.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages13-23
Number of pages11
DOIs
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
Volume2303
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Chemical shift perturbation
  • Dissociation constant
  • Glycosaminoglycan
  • Heparan sulfate
  • Heparin
  • Nuclear magnetic resonance (NMR)
  • Protein–ligand interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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