Proteinâ??ligand titrations can readily be monitored with a trimethylsilyl (TMS) tag. Owing to the intensity, narrow line shape and unique chemical shift of a TMS group, dissociation constants can be determined from straightforward 1D 1H-NMR spectra not only in the fast but also in the slow exchange limit. The tag is easily attached to cysteine residues and a sensitive reporter of ligand binding also at sites where it does not interfere with ligand binding or catalytic efficiency of the target protein. Its utility is demonstrated for the Zika virus NS2Bâ??NS3 protease and the human prolyl isomerase FK506 binding protein.
Proteinâ??Ligand Interactions, Volume 53. Methods and Principles in Medicinal Chemistry â?? Biotechnology, Healthcare ... - satPRnews (press release)
Proteinâ??Ligand Interactions, Volume 53. Methods and Principles in Medicinal Chemistry â?? Biotechnology, Healthcare ... - satPRnews (press release)
<img alt="" height="1" width="1">
Proteinâ??Ligand Interactions, Volume 53. Methods and Principles in Medicinal Chemistry â?? Biotechnology, Healthcare ...
satPRnews (press release)
The first part provides a basic understanding of the factors governing proteinâ??ligand interactions, followed by a comparison of key experimental methods (calorimetry, surface plasmon resonance, NMR) used in generating interaction data. The second half ...
...
nmrlearner
Online News
0
01-23-2018 02:57 AM
Target-specific NMR detection of proteinâ??ligand interactions with antibody-relayed 15 N-group selective STD
Target-specific NMR detection of proteinâ??ligand interactions with antibody-relayed 15 N-group selective STD
Abstract
Fragment-based drug design has been successfully applied to challenging targets where the detection of the weak proteinâ??ligand interactions is a key element. 1H saturation transfer difference (STD) NMR spectroscopy is a powerful technique for this work but it requires pure homogeneous proteins as targets. Monoclonal antibody (mAb)-relayed 15N-GS STD spectroscopy has been developed to resolve the problem of protein mixtures and...
nmrlearner
Journal club
0
11-25-2016 02:22 AM
Determination of ligand binding modes in weak proteinâ??ligand complexes using sparse NMR data
Determination of ligand binding modes in weak proteinâ??ligand complexes using sparse NMR data
Abstract
We describe a general approach to determine the binding pose of small molecules in weakly bound proteinâ??ligand complexes by deriving distance constraints between the ligand and methyl groups from all methyl-containing residues of the protein. We demonstrate that using a single sample, which can be prepared without the use of expensive precursors, it is possible to generate high-resolution data rapidly and obtain the resonance assignments of...
nmrlearner
Journal club
0
11-19-2016 08:35 PM
[NMR paper] Overview of Probing Protein-Ligand Interactions Using NMR.
Overview of Probing Protein-Ligand Interactions Using NMR.
Overview of Probing Protein-Ligand Interactions Using NMR.
Curr Protoc Protein Sci. 2015;81:17.18.1-17.18.24
Authors: Aguirre C, Cala O, Krimm I
Abstract
Nuclear magnetic resonance (NMR) is a powerful technique for the study and characterization of protein-ligand interactions. In this unit we review both experiments where the NMR spectrum of the protein is observed (protein-observed NMR experiments) and those where the NMR spectra of the ligand is observed...
nmrlearner
Journal club
0
08-04-2015 03:00 PM
Proteinâ??ligand structure guided by backbone and side-chain proton chemical shift perturbations
Proteinâ??ligand structure guided by backbone and side-chain proton chemical shift perturbations
Abstract
The fragment-based drug design approach consists of screening libraries of fragment-like ligands, to identify hits that typically bind the protein target with weak affinity ( \(100\,\upmu \hbox {M}\) â??5Â*mM). The determination of the proteinâ??fragment complex 3D structure constitutes a crucial step for uncovering the key interactions responsible for...
nmrlearner
Journal club
0
09-26-2014 01:03 PM
Accuracy and precision of proteinâ??ligand interaction kinetics determined from chemical shift titrations
Accuracy and precision of proteinâ??ligand interaction kinetics determined from chemical shift titrations
Abstract NMR-monitored chemical shift titrations for the study of weak proteinâ??ligand interactions represent a rich source of information regarding thermodynamic parameters such as dissociation constants (K D ) in the micro- to millimolar range, populations for the free and ligand-bound states, and the kinetics of interconversion between states, which are typically within the fast exchange regime on the NMR timescale. We recently developed two chemical shift titration methods...
nmrlearner
Journal club
0
10-24-2012 10:28 PM
Increased precision for analysis of proteinâ??ligand dissociation constants determined from chemical shift titrations
Increased precision for analysis of proteinâ??ligand dissociation constants determined from chemical shift titrations
Abstract NMR is ideally suited for the analysis of proteinâ??protein and protein ligand interactions with dissociation constants ranging from ~2 μM to ~1 mM, and with kinetics in the fast exchange regime on the NMR timescale. For the determination of dissociation constants (K D ) of 1:1 proteinâ??protein or proteinâ??ligand interactions using NMR, the protein and ligand concentrations must necessarily be similar in magnitude to the K D , and nonlinear least squares...
nmrlearner
Journal club
0
05-01-2012 07:06 AM
[NMR paper] Probing the binding entropy of ligand-protein interactions by NMR.
Probing the binding entropy of ligand-protein interactions by NMR.
Related Articles Probing the binding entropy of ligand-protein interactions by NMR.
Chembiochem. 2005 Sep;6(9):1585-91
Authors: Homans SW
Trimethylsilyl tag for probing proteinâ??ligand interactions by NMR
Linear Mode
