NMR studies on carbohydrate interactions with DC-SIGN towards a quantitative STD analysis

The recent introduction of saturation transfer difference (STD) NMR has increased the tools for the study of protein–carbohydrate complexes. This is useful when it is combined with transfer nuclear Overhauser enhancement spectroscopy (NOESY) measurement, or when it is interpreted using the expected calculated values of transference, yielding additional, very valuable information for the study of this type of complex. The objective of this work is to cover the advances of the STD technique as exemplified by the investigations of DC-SIGN (dendritic cell-specific ICAM-3 grabbing non-integrin) recognition by simple carbohydrates or mimics of them, based on structures containing a terminal mannose or fucose. We also will discuss the methods for quantification of the STD values based on the initial growing rates with the saturation time.

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Understanding heat driven gelation of anionic cellulose nanofibrils: Combining saturation transfer difference (STD) NMR, small angle X-ray scattering (SAXS) and rheology

Journal of Colloid and Interface Science ◽

10.1016/j.jcis.2018.09.085 ◽

2019 ◽

Vol 535 ◽

pp. 205-213 ◽

Cited By ~ 11

Author(s):

Vincenzo Calabrese ◽

Juan C. Muñoz-García ◽

Julien Schmitt ◽

Marcelo A. da Silva ◽

Janet L. Scott ◽

...

Keyword(s):

Small Angle ◽

Cellulose Nanofibrils ◽

Saturation Transfer ◽

X Ray ◽

X Ray Scattering ◽

Saturation Transfer Difference ◽

Std Nmr ◽

Ray Scattering

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Saturation transfer difference NMR on the integral trimeric membrane transport protein GltPh determines cooperative substrate binding

Scientific Reports ◽

10.1038/s41598-020-73443-z ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Jenny L. Hall ◽

Azmat Sohail ◽

Eurico J. Cabrita ◽

Colin Macdonald ◽

Thomas Stockner ◽

...

Keyword(s):

Membrane Transport ◽

Excitatory Amino Acid ◽

Transport Protein ◽

Amino Acid Transporters ◽

Binding Affinities ◽

Saturation Transfer ◽

Saturation Transfer Difference ◽

Membrane Transport Protein ◽

The Central Nervous System ◽

Std Nmr

Abstract Saturation-transfer difference (STD) NMR spectroscopy is a fast and versatile method which can be applied for drug-screening purposes, allowing the determination of essential ligand binding affinities (KD). Although widely employed to study soluble proteins, its use remains negligible for membrane proteins. Here the use of STD NMR for KD determination is demonstrated for two competing substrates with very different binding affinities (low nanomolar to millimolar) for an integral membrane transport protein in both detergent-solubilised micelles and reconstituted proteoliposomes. GltPh, a homotrimeric aspartate transporter from Pyrococcus horikoshii, is an archaeal homolog of mammalian membrane transport proteins—known as excitatory amino acid transporters (EAATs). They are found within the central nervous system and are responsible for fast uptake of the neurotransmitter glutamate, essential for neuronal function. Differences in both KD’s and cooperativity are observed between detergent micelles and proteoliposomes, the physiological implications of which are discussed.

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Saturation Transfer Difference in Characterization of Glycosaminoglycan-Protein Interactions

SLAS TECHNOLOGY Translating Life Sciences Innovation ◽

10.1177/2472630320921130 ◽

2020 ◽

Vol 25 (4) ◽

pp. 307-319

Author(s):

William P. Vignovich ◽

Vitor H. Pomin

Keyword(s):

Protein Interactions ◽

Specific Binding ◽

Coagulation Factors ◽

Sulfated Polysaccharides ◽

Saturation Transfer ◽

Intermolecular Complexes ◽

Saturation Transfer Difference ◽

Protein Partners ◽

Preferential Binding ◽

Std Nmr

Novel methods in nuclear magnetic resonance (NMR) spectroscopy have recently been developed to investigate the binding properties of intermolecular complexes endowed with biomedical functions. Among these methods is the saturation transfer difference (STD), which enables the mapping of specific binding motifs of functional ligands. STD can efficiently uncover the specific and preferential binding sites of these ligands in their intermolecular complexes. This is particularly useful in the case of glycosaminoglycans (GAGs), a group of sulfated polysaccharides that play pivotal roles in various biological and pathological processes. The activity of GAGs is ultimately mediated through molecular interactions with key functional proteins, namely, GAG-binding proteins (GBPs). The quality of the GAG-GBP interactions depends on sulfation patterns, oligosaccharide length, and the composing monosaccharides of GAGs. Through STD NMR, information about the atoms of the GAG ligands involved in the complexes is provided. Here we highlight the latest achievements of the literature using STD NMR on GAG oligosaccharide-GBP complexes. Interestingly, most of the GBPs studied so far by STD NMR belong to one of the three major classes: coagulation factors, growth factors, or chemokine/cytokines. Unveiling the structural requirements of GAG ligands in bindings with their protein partners is a crucial step to understand the biochemical and medical actions of GAGs. This process is also a requirement in GAG-based drug discovery and development.

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