TY - CHAP
T1 - Solution NMR Spectroscopy for Characterizing Protein–Glycosaminoglycan Interactions
AU - Joseph, Prem Raj B.
AU - Sepuru, Krishna Mohan
AU - Poluri, Krishna Mohan
AU - Rajarathnam, Krishna
N1 - Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - 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.
AB - 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.
KW - Chemical shift perturbation
KW - Dissociation constant
KW - Glycosaminoglycan
KW - Heparan sulfate
KW - Heparin
KW - Nuclear magnetic resonance (NMR)
KW - Protein–ligand interactions
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U2 - 10.1007/978-1-0716-1398-6_2
DO - 10.1007/978-1-0716-1398-6_2
M3 - Chapter
C2 - 34626366
AN - SCOPUS:85117111067
T3 - Methods in Molecular Biology
SP - 13
EP - 23
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -