A5059511896- Advanced NMR Techniques and Applications
- NMR spectroscopy and applications
- Solid-state spectroscopy and crystallography
- Protein Structure and Dynamics
- Trypanosoma species research and implications
- Research on Leishmaniasis Studies
- Enzyme Structure and Function
- Hippo pathway signaling and YAP/TAZ
- Protein Kinase Regulation and GTPase Signaling
- Photosynthetic Processes and Mechanisms
- Muon and positron interactions and applications
- RNA Research and Splicing
- Biochemical and Molecular Research
- Heat shock proteins research
- Electron Spin Resonance Studies
- Advanced MRI Techniques and Applications
- Ubiquitin and proteasome pathways
- RNA and protein synthesis mechanisms
- Cancer-related gene regulation
- Atomic and Subatomic Physics Research
- Genetics and Neurodevelopmental Disorders
- Signaling Pathways in Disease
- Enzyme Catalysis and Immobilization
- Microbial metabolism and enzyme function
- Microbial Metabolic Engineering and Bioproduction
Leibniz-Forschungsinstitut für Molekulare Pharmakologie
2013-2025
Forschungsverbund Berlin
2021
Leibniz Association
2014
Leibniz Institute for Neurobiology
2006-2013
Charité - Universitätsmedizin Berlin
2007
Massachusetts Institute of Technology
2003
AstraZeneca (Sweden)
2003
Freie Universität Berlin
2003
Technische Universität Berlin
1998-2000
Max Delbrück Center
2000
Slim peaks: Using a perdeuterated protein recrystallized from 10:90 H2O:D2O mixture in magic-angle spinning (MAS) solid-state NMR spectroscopy experiments gives small 1H line widths at moderate frequencies without application of homonuclear decoupling. This labeling strategy opens new perspectives for assignment large spin systems. Structure investigations biological solids by high- resolution has rapidly progressed the last few years and resulted complete structure elucidation several...
Microorganisms form surface-attached communities, termed biofilms, which can serve as protection against host immune reactions or antibiotics. Bacillus subtilis biofilms contain TasA major proteinaceous component in addition to exopolysaccharides. In stark contrast the initially unfolded biofilm proteins of other bacteria, is a soluble, stably folded monomer, whose structure we have determined by X-ray crystallography. Subsequently, characterized vitro different oligomeric forms NMR, EM,...
Abstract Microtubules are filamentous structures necessary for cell division, motility and morphology, with dynamics critically regulated by microtubule-associated proteins (MAPs). Here we outline the molecular mechanism which MAP, COMPANION OF CELLULOSE SYNTHASE1 (CC1), controls microtubule bundling to sustain plant growth under salt stress. CC1 contains an intrinsically disordered N-terminus that links microtubules at evenly distributed points through four conserved hydrophobic regions. By...
In this paper, a three-dimensional (3D) NMR-based approach for the determination of fold moderately sized proteins by solid-state magic-angle spinning (MAS) NMR is presented and applied to alpha-spectrin SH3 domain. This methodology includes measurement multiple (13)C-(13)C distance restraints on biosynthetically site-directed (13)C-enriched samples, obtained growing bacteria [2-(13)C]glycerol [1,3-(13)C]glycerol. 3D (15)N-(13)C-(13)C dipolar correlation experiments were resolve overlap...
In this communication, we demonstrate the feasibility of 1H detection in MAS solid-state NMR for a microcrystalline, uniformly 2H,15N-labeled sample SH3 domain chicken α-spectrin, using pulsed field gradients suppression water magnetization. Today, B0 are employed routinely solution-state coherence order selection and solvent suppression. We suggest to use purge magnetization which cannot be suppressed conventional schemes. The achievable gain sensitivity is 5 compared 15N detected version...
MAS solid-state NMR experiments applied to biological solids are still hampered by low sensitivity and resolution. In this work, we employ a deuteration scheme in which individual methyl groups selectively protonated. This labeling allows the acquisition of proton carbon correlation spectra with resolution comparable that solution-state experiments. We observe an increase factor 10-15 compared standard heteronuclear using PMLG for 1H,1H dipolar decoupling indirect dimension. At same time,...
Abstract Cross‐linking mass spectrometry (XL‐MS) is an attractive method for the proteome‐wide characterization of protein structures and interactions. Currently, depth in vivo XL‐MS studies lagging behind established applications to cell lysates, because cross‐linking reagents that can penetrate intact cells strategies enrich cross‐linked peptides lack efficiency. To tackle these limitations, we have developed a phosphonate‐containing cross‐linker, tBu‐PhoX, efficiently permeates various...
Channelrhodopsins (ChR) are light-gated ion channels of green algae that widely used to probe the function neuronal cells with light. Most ChRs show a substantial reduction in photocurrents during illumination, process named "light adaptation". The main objective this spectroscopic study was elucidate molecular processes associated light-dark adaptation. Here we by liquid and solid-state nuclear magnetic resonance spectroscopy retinal chromophore fully dark-adapted ChR is exclusively an...
ABSTRACT The antigen 85 (Ag85) protein family, consisting of Ag85A, -B, and -C, is vital for Mycobacterium tuberculosis due to its role in cell envelope biogenesis. mycoloyl transferase activity these proteins generates trehalose dimycolate (TDM), an lipid essential M. virulence, wall arabinogalactan-linked mycolic acids. Inhibition enzymes through substrate analogs hinders growth mycobacteria, but a link acid synthesis has not been established. In this study, we characterized novel...
Studying mechanisms of bacterial biofilm generation is vital importance to understanding cell-cell communication, multicellular cohabitation principles, and the higher resilience microorganisms in a against antibiotics. Biofilms nonpathogenic, gram-positive soil bacterium
Many MAS (magic angle spinning) solid-state NMR investigations of biologically relevant protein samples are hampered by poor resolution, particularly in the 15N chemical shift dimension. We show that dynamics nanosecond−microsecond time scale can induce significant line broadening resonances experiments. Averaging 15NHα/β multiplet components due to 1H decoupling induces effective relaxation coherence case N−H spin pair undergoes motion. High resolution spectra then only be recorded...
In this paper, we present the measurement of (15)N-T(1) relaxation times in solid state for a perdeuterated protein which exchangeable protons are back substituted during recrystallization using buffer contains 10% H(2)O and 90% D(2)O. We find large variations (15)N time, even within same beta sheet. By comparing measured protonated deuterated (using above mentioned approach), conclude that (1)H driven (15)N,(15)N spin diffusion has significant impact on absolute time proteins. This effect...
Dynamic nuclear polarization exploits electron spin to boost signal-to-noise in magic-angle-spinning (MAS) NMR, creating new opportunities materials science, structural biology, and metabolomics studies. Since protein NMR spectra recorded under DNP conditions can show improved spectral resolution at 180-200 K compared 110 K, we investigate the effects of AMUPol various deuterated TOTAPOL isotopologues on sensitivity these temperatures, using proline reproducibly prepared SH3 domain samples....
Abstract Solution NMR is typically applied to biological systems with molecular weights < 40 kDa whereas magic-angle-spinning (MAS) solid-state traditionally targets very large, oligomeric proteins and complexes exceeding 500 in mass, including fibrils crystalline protein preparations. Here, we propose that the gap between these size regimes can be filled by approach presented enables investigation of soluble fully protonated range 40–140 kDa. As a key step, ultracentrifugation produces...
In this communication, we demonstrate the feasibility of obtaining long-range 1H−1H distance information by MAS solid-state NMR for a microcrystalline, uniformly 2H,15N-labeled sample SH3 domain chicken α-spectrin. The experiments yield NOESY-type spectra and rely on favorable dispersion 15N chemical shifts protein backbone. Perdeuteration nonexchangeable sites is employed to simplify proton spin systems obtain multiple structural information. Two mixing schemes, double quantum filtered...
We show in this communication that dynamic information for uniformly 2H,13C,15N isotopically enriched, crystalline proteins can be obtained by MAS solid-state NMR spectroscopy. The experiments make use of the deuterium quadrupolar tensor, which is dominant interaction mechanism. Dynamic properties are accessed measurement size coupling constant, Cq, and value asymmetry parameter, η, via evolution chemical shift, as well T1 relaxation times. Three-dimensional performed order to obtain...
Rapid advances in solid-state MAS NMR made it possible to probe protein dynamics on a per-residue basis, similar solution experiments. In this work we compare methyl 2H relaxation rates measured the solid and liquid samples of alpha-spectrin SH3 domain. The data are treated using model-free approach separate contributions from overall molecular tumbling fast internal motion. latter part forms basis for comparison with data. Although accuracy measurements is limited by deuterium spin...
Abstract Shank is the central scaffolding protein of postsynaptic density (PSD) complex found in cells nervous system. Cellular studies indicate a prominent role organization PSD, development neuronal morphology, signaling, and synaptic plasticity, thus linking functions to molecular basis learning memory. Mutations gene have been several disorders including mental retardation, typical autism, Asperger syndrome. linked PSD via its PDZ domain that binds C‐terminus guanylate‐kinase‐associated...