Characterizing protein–glycosaminoglycan interactions using solution NMR spectroscopy

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

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Solution nuclear magnetic resonance (NMR) spectroscopy and, in particular, chemical shift perturbation (CSP) titration experiments are ideally suited for characterizing the binding interface of macromolecular complexes. 1 H- 15 N-HSQC-based CSP studies have become the method of choice due to their simplicity, short time requirements, and not requiring high-level NMR expertise. Nevertheless, CSP studies for characterizing protein–glycosaminoglycan (GAG) interactions have been challenging due to binding-induced aggregation/precipitation and/or poor quality data. In this chapter, we discuss how optimizing experimental variables such as protein concentration, GAG size, and sensitivity of NMR instrumentation can overcome these roadblocks to obtain meaningful structural insights into protein–GAG interactions.

Original languageEnglish (US)
Pages (from-to)325-333
Number of pages9
JournalMethods in Molecular Biology
Volume1229
DOIs
StatePublished - Jan 1 2015
Externally publishedYes

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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