A5081203545- Advanced NMR Techniques and Applications
- Protein Structure and Dynamics
- NMR spectroscopy and applications
- Heat shock proteins research
- Solid-state spectroscopy and crystallography
- Enzyme Structure and Function
- Electron Spin Resonance Studies
- Connexins and lens biology
- Computational Drug Discovery Methods
- Spectroscopy and Quantum Chemical Studies
- RNA and protein synthesis mechanisms
- Mass Spectrometry Techniques and Applications
- RNA modifications and cancer
- Photosynthetic Processes and Mechanisms
- RNA Research and Splicing
- Receptor Mechanisms and Signaling
- History and advancements in chemistry
- Advanced MRI Techniques and Applications
- Atomic and Subatomic Physics Research
- Nanopore and Nanochannel Transport Studies
- Crystallography and Radiation Phenomena
- Biochemical effects in animals
- DNA and Nucleic Acid Chemistry
- Muon and positron interactions and applications
- Calcium signaling and nucleotide metabolism
Technical University of Munich
2010-2024
Center for Integrated Protein Science Munich
2013-2021
Helmholtz Zentrum München
2010-2019
Leibniz-Forschungsinstitut für Molekulare Pharmakologie
2011-2012
Goethe University Frankfurt
2009
Biological magic angle spinning (MAS) solid-state nuclear magnetic resonance spectroscopy has developed rapidly over the past two decades. For structure determination of a protein by NMR, routinely 13C,13C distance restraints as well dihedral are employed. In protonated samples, this is achieved growing bacterium on medium which contains [1,3]-13C glycerol or [2]-13C to dilute 13C spin system. Labeling schemes, rely heteronuclei, insensitive both for detection and in terms quantification...
HuR/ELAVL1 is an RNA-binding protein involved in differentiation and stress response that acts primarily by stabilizing messenger RNA (mRNA) targets. HuR comprises three recognition motifs (RRMs) where the structure binding of RRM3 full-length remain poorly understood. Here, we report crystal structures free bound to cognate RNAs. Our structural, NMR biochemical data show mediates canonical interactions reveal molecular details a dimerization interface localized on α-helical face RRM3. SAXS...
Hsp90 is a molecular chaperone that interacts with specific set of client proteins and assists their folding. The underlying mechanisms, involving dynamic transitions between open closed conformations, are still enigmatic. Combining nuclear magnetic resonance, small-angle x-ray scattering, biochemical experiments, we have identified key intermediate state induced by adenosine triphosphate (ATP) binding, in which rotation the N-terminal domain (NTD) yields arrangement poised for closing. This...
Both protonated and deuterated samples were employed in the study of L7Ae box C/D RNA complex by 1H-detected solid-state NMR spectroscopy. This approach yielded high-resolution spectra was used to determine intermolecular interface extract structural parameters with high accuracy. As a service our authors readers, this journal provides supporting information supplied authors. Such materials are peer reviewed may be re-organized for online delivery, but not copy-edited or typeset. Technical...
MAS solid-state NMR is capable of determining structures protonated solid proteins using proton-detected experiments. These experiments are performed at rotation frequency around 110 kHz, employing 0.5 mg material. Here, we compare 1H, 13C correlation spectra obtained from and deuterated microcrystalline 111 show that the spectral quality samples superior to those acquired in terms resolution sensitivity. In comparison samples, yield a gain on order 3 2 proton carbon dimensions,...
In the last decade, proton detection in magic-angle spinning (MAS) solid-state NMR became a popular strategy for biomolecular structure determination. particular, probe technology has experienced tremendous progress with smaller and diameter rotors achieving ever higher MAS frequencies. rotation frequencies beyond 100 kHz allow to observe assign protons fully protonated samples. these experiments, resolution is however compromised as homogeneous proton–proton dipolar coupling interactions...
We introduce a labeling scheme for magic angle spinning (MAS) solid-state NMR that is based on deuteration in combination with dilution of the carbon spin system. The strategy achieves spectral editing by simplification HαCα and aliphatic side chain region. A reduction both proton density fast (≥50 kHz) essential to retrieve artifact-free (13)C-R1 relaxation data carbons. obtain good agreement between experimental order parameters extracted from molecular dynamics (MD) trajectory, which...
The measurement of dipolar couplings among directly bonded nuclei yields direct information on the amplitude dynamic processes in solid-state. For a reliable motional analysis using, e.g., model-free approach, correct quantification absolute values these order parameters is absolutely essential. In absence reference value for rigid limit, too low coupling might be misinterpreted as motion. Therefore, detailed understanding effects that influence experimental necessary. We compare here REDOR...
NMR spectroscopy at ultra-high magnetic fields requires improved radiofrequency (rf) pulses to cover the increased spectral bandwidth. Optimized 90° pulse pairs were introduced as Ramsey-type cooperative (Ram-COOP) for biomolecular applications. The Ram-COOP element provides broadband excitation with enhanced sensitivity and reduced artifacts even >1.0 GHz 1 H Larmor frequency (23 T). A pair of 30 μs achieves an bandwidth 100 kHz a maximum rf field 20 kHz, more than three-fold compared by...
The recently discovered SAFit class of inhibitors against the Hsp90 co-chaperone FKBP51 show greater than 10 000-fold selectivity over its closely related paralogue FKBP52. However, mechanism underlying this remained unknown. By combining NMR spectroscopy, biophysical and computational methods with mutational analysis, we that molecules bind to a transient pocket in FKBP51. This represents weakly populated conformation resembling inhibitor-bound state FKBP51, suggesting conformational...
A notable drawback of NMR spectroscopy is its inherently low sensitivity: 95% the measuring time consists solely idle delays during which nuclei regain their Boltzmann equilibrium. Here, a strategy for solid-state 13C experiments presented that allows user to acquire spectra in periods are notably shorter than previously necessary. Experiments band-selective nature may utilize cooling potential unperturbed lower spin temperature excited neighbors. As we demonstrate, it becomes possible...
Abstract We recently introduced RAP (reduced adjoining protonation) labelling as an easy to implement and cost-effective strategy yield selectively methyl protonated protein samples. show here that even though the amount of H 2 O employed in bacterial growth medium is rather low, intensities obtained MAS solid-state NMR 1 H, 13 C correlation spectra are comparable for samples which α-ketoisovalerate was precursor. In addition correlations Leu Val residues, labelled also resonances all...
Abstract Soluble proteins are universally packed with a hydrophobic core and polar surface that drive the protein folding process. Yet charged networks within central often indispensable for biological function. Here, we show natural buried ion-pairs stabilised by amphiphilic residues electrostatically shield motif from its surroundings to gain structural stability. To explore this effect, build artificial combining directed computational design biophysical experiments. Our findings...
Abstract NMR spectroscopy at ultra‐high magnetic fields requires improved radiofrequency ( rf ) pulses to cover the increased spectral bandwidth. Optimized 90° pulse pairs were introduced as Ramsey‐type cooperative (Ram‐COOP) for biomolecular applications. The Ram‐COOP element provides broadband excitation with enhanced sensitivity and reduced artifacts even >1.0 GHz 1 H Larmor frequency (23 T). A pair of 30 μs achieves an bandwidth 100 kHz a maximum field 20 kHz, more than three‐fold...
Strukturaufklärung von Protein-RNA-Komplexen: Die 1H-detektierte Untersuchung des L7Ae-Box-C/D-RNA-Komplexes gelang durch Festkörper-NMR-Spektroskopie mithilfe hochaufgelöster Spektren. Methode ermöglicht die hoch genaue Bestimmung Strukturparametern und intermolekularen Interaktionsstellen. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed may be re-organized for online delivery, but not...
Abstract The recently discovered SAFit class of inhibitors against the Hsp90 co‐chaperone FKBP51 show greater than 10 000‐fold selectivity over its closely related paralogue FKBP52. However, mechanism underlying this remained unknown. By combining NMR spectroscopy, biophysical and computational methods with mutational analysis, we that molecules bind to a transient pocket in FKBP51. This represents weakly populated conformation resembling inhibitor‐bound state FKBP51, suggesting...