In recent years, we developed a toolbox of heavy isotope containing compounds, which serve as metabolic amino acid precursors in the E. coli-based overexpression of aromatic residue labeled proteins. Our labeling techniques show excellent results both in terms of selectivity and isotope incorporation levels. They are additionally distinguished by low sample production costs and meet the economic demands to further implement protein NMR spectroscopy as a routinely used method in drug development processes. Different isotopologues allow for the assembly of optimized protein samples, which fulfill the requirements of various NMR experiments to elucidate protein structures, analyze conformational dynamics, or probe interaction surfaces. In the present article, we want to summarize the precursors we developed so far and give examples of their special value in the probing of proteinā??ligand interaction.
Anthranilic acid, the new player in the ensemble of aromatic residue labeling precursor compounds
Anthranilic acid, the new player in the ensemble of aromatic residue labeling precursor compounds
Abstract
The application of metabolic precursors for selective stable isotope labeling of aromatic residues in cell-based protein overexpression has already resulted in numerous NMR probes to study the structural and dynamic characteristics of proteins. With anthranilic acid, we present the structurally simplest precursor for exclusive tryptophan side chain labeling. A synthetic route to 13C, 2H isotopologues allows the installation of isolated 13Cā??1H...
nmrlearner
Journal club
0
08-31-2017 01:22 PM
Highly efficient residue -selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain
Highly efficient residue -selective labeling with isotope-labeled Ile, Leu, and Val using a new auxotrophic E. coli strain
Abstract
We recently developed a practical protocol for preparing proteins bearing stereo-selectively 13C-methyl labeled leucines and valines, instead of the commonly used 13C-methyl labeled precursors for these amino acids, by E. coli cellular expression. Using this protocol, proteins with any combinations of isotope-labeled or unlabeled Leu and Val residues were prepared, including some that could not be prepared by the...
nmrlearner
Journal club
0
06-07-2016 02:39 PM
[NMR paper] NMR analysis of cross strand aromatic interactions in an 8 residue hairpin and a 14 residue three stranded ?-sheet peptide.
NMR analysis of cross strand aromatic interactions in an 8 residue hairpin and a 14 residue three stranded ?-sheet peptide.
http://www.bionmr.com//www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--pubs.acs.org-images-pubmed-acspubs.jpg Related Articles NMR analysis of cross strand aromatic interactions in an 8 residue hairpin and a 14 residue three stranded ?-sheet peptide.
J Phys Chem B. 2012 Dec 13;116(49):14207-15
Authors: Sonti R, Rai R, Ragothama S, Balaram P
Abstract
Cross strand aromatic interactions between a facing pair of...
nmrlearner
Journal club
0
06-13-2013 06:14 PM
[NMR paper] pH-Dependent Conformation, Dynamics, and Aromatic Interaction of*the*Gating Tryptophan Residue of the Influenza M2 Proton Channel from*Solid-State NMR.
pH-Dependent Conformation, Dynamics, and Aromatic Interaction of*the*Gating Tryptophan Residue of the Influenza M2 Proton Channel from*Solid-State NMR.
Related Articles pH-Dependent Conformation, Dynamics, and Aromatic Interaction of*the*Gating Tryptophan Residue of the Influenza M2 Proton Channel from*Solid-State NMR.
Biophys J. 2013 Apr 16;104(8):1698-708
Authors: Williams JK, Zhang Y, Schmidt-Rohr K, Hong M
Abstract
The M2 protein of the influenza virus conducts protons into the virion under external acidic pH. The proton selectivity of...
nmrlearner
Journal club
0
04-23-2013 08:37 PM
Hyperpolarized (13) C-labelled anhydrides as DNP precursors of metabolic MRI agents
From the The DNP-NMR Blog:
Hyperpolarized (13) C-labelled anhydrides as DNP precursors of metabolic MRI agents
Colombo Serra, S., et al., Hyperpolarized 13C-labelled anhydrides as DNP precursors of metabolic MRI agents. Contrast Media & Molecular Imaging, 2012. 7(5): p. 469-477.
http://www.ncbi.nlm.nih.gov/pubmed/22821881
nmrlearner
News from NMR blogs
0
04-15-2013 08:52 AM
Alternative SAIL-Trp for robust aromatic signal assignment and determination of the Ļ?2 conformation by intra-residue NOEs
Alternative SAIL-Trp for robust aromatic signal assignment and determination of the Ļ?2 conformation by intra-residue NOEs
Abstract Tryptophan (Trp) residues are frequently found in the hydrophobic cores of proteins, and therefore, their side-chain conformations, especially the precise locations of the bulky indole rings, are critical for determining structures by NMR. However, when analyzing -proteins, the observation and assignment of the ring signals are often hampered by excessive overlaps and tight spin couplings. These difficulties have been greatly alleviated by using...
nmrlearner
Journal club
0
09-27-2011 07:04 AM
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
Frequency-selective heteronuclear dephasing and selective carbonyl labeling to deconvolute crowded spectra of membrane proteins by magic angle spinning NMR.
J Magn Reson. 2011 Mar 17;
Authors: Traaseth NJ, Veglia G
We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR...
nmrlearner
Journal club
0
04-13-2011 11:57 PM
Frequency-Selective Heteronuclear Dephasing and Selective Carbonyl Labeling to Deconvolute Crowded Spectra of Membrane Proteins By Magic Angle Spinning NMR
Frequency-Selective Heteronuclear Dephasing and Selective Carbonyl Labeling to Deconvolute Crowded Spectra of Membrane Proteins By Magic Angle Spinning NMR
Publication year: 2011
Source: Journal of Magnetic Resonance, In Press, Accepted Manuscript, Available online 17 March 2011</br>
Nathaniel J., Traaseth , Gianluigi, Veglia</br>
We present a new method that combines carbonyl-selective labeling with frequency-selective heteronuclear recoupling to resolve the spectral overlap of magic angle spinning (MAS) NMR spectra of membrane proteins in fluid lipid membranes with broad lines and...
Late metabolic precursors for selective aromatic residue labeling
Linear Mode
