Characterizing protein–glycosaminoglycan interactions using solution NMR spectroscopy

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

    Research output: Contribution to journalArticlepeer-review

    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 - 2015

    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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