title: Characterizing protein-glycosaminoglycan interactions using solution NMR spectroscopy.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555-1055, USA.
authors.forename: Prem Raj B
authors.initials: PR
authors.lastname: Joseph
authors.forename: Krishna Mohan
authors.initials: KM
authors.lastname: Poluri
authors.forename: Krishna Mohan
authors.initials: KM
authors.lastname: Sepuru
authors.forename: Krishna
authors.initials: K
authors.lastname: Rajarathnam
mesh_terms: D018925:Chemokines / Q000737:chemistry / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions; D013223:Statistics as Topic; D017186:Titrimetry
chemical_list: D018925:Chemokines; D006025:Glycosaminoglycans; D011506:Proteins; D012996:Solutions