A5014424497- Advanced NMR Techniques and Applications
- Protein Structure and Dynamics
- Lipid Membrane Structure and Behavior
- NMR spectroscopy and applications
- Enzyme Structure and Function
- Solid-state spectroscopy and crystallography
- Advanced MRI Techniques and Applications
- Influenza Virus Research Studies
- Parkinson's Disease Mechanisms and Treatments
- Electron Spin Resonance Studies
- Muon and positron interactions and applications
- Mass Spectrometry Techniques and Applications
- Alzheimer's disease research and treatments
- Atomic and Subatomic Physics Research
- Physics of Superconductivity and Magnetism
- Genomics and Chromatin Dynamics
- Neurological disorders and treatments
- Bacteriophages and microbial interactions
- Molecular Sensors and Ion Detection
- Ion channel regulation and function
- RNA Research and Splicing
- Nuclear Physics and Applications
- Galectins and Cancer Biology
- Protein Tyrosine Phosphatases
- Radio Astronomy Observations and Technology
Max Planck Institute for the Study of Religious and Ethnic Diversity
2025
Max Planck Institute for Multidisciplinary Sciences
2022-2025
University of Delaware
2022-2024
University of Pittsburgh
2024
Max Planck Institute for Biophysical Chemistry
2018-2022
Institut de Pharmacologie et de Biologie Structurale
2018
Ludwig-Maximilians-Universität München
2017
The available magnetic field strength for high resolution NMR in persistent superconducting magnets has recently improved from 23.5 to 28 Tesla, increasing the proton resonance frequency 1 1.2 GHz. For magic-angle spinning (MAS) NMR, this is expected improve resolution, provided sample preparation results homogeneous broadening. We compare two-dimensional (2D) detected MAS spectra of four membrane proteins at 950 and 1200 MHz. find a consistent improvement that scales superlinearly with...
Significance Here we show how AlkL, a minimalistic outer-membrane protein from oil-consuming bacteria, exploits dynamics of extracellular loops to channel substrate across the polysaccharide barrier and into hydrophobic interior outer membrane. This work represents unique example side-by-side atomic-level structure determination by solution NMR in detergents solid-state lipid bilayers, which critically demonstrates importance environment investigate function. Corroborating our experimental...
Aggregation intermediates of Parkinson’s disease–related α-synuclein on liposomes are structurally characterized by NMR.
The nucleocapsid (N) protein is one of the four structural proteins SARS-CoV-2 virus and plays a crucial role in viral genome organization and, hence, replication pathogenicity. N-terminal domain (NNTD) binds to genomic RNA thus comprises potential target for inhibitor vaccine development. We determined atomic-resolution structure crystalline NNTD by integrating solid-state magic angle spinning (MAS) NMR X-ray diffraction. Our combined approach provides atomic details packing interfaces as...
The arrangement of histidine side chains in influenza A M2 tetramer determines their pKa values, which define pH-controlled proton conduction critical to the virus lifecycle. Both water-associated and hydrogen-bonded imidazole–imidazolium quaternary structures have been proposed, based on crystal NMR chemical shifts, respectively. Here we show, using domain construct lipid bilayers, that imidazole rings are hydrogen bonded even at a pH 7.8 neutral charge state. An intermolecular 8.9 ± 0.3 Hz...
The voltage-dependent anion channel (VDAC), the most abundant protein in outer mitochondrial membrane, is responsible for transport of all ions and metabolites into out mitochondria. Larger than any β-barrel structures determined to date by magic-angle spinning (MAS) NMR, but smaller size limit cryo-electron microscopy (cryo-EM), VDAC1's 31 kDa has long been a bottleneck determining its structure near-native lipid bilayer environment. Using single two-dimensional (2D) crystalline sample...
Aggregation intermediates play a pivotal role in the assembly of amyloid fibrils, which are central to pathogenesis neurodegenerative diseases. The structures filamentous and mature fibrils now efficiently determined by single-particle cryo-electron microscopy. By contrast, smaller pre-fibrillar α-Synuclein (αS) oligomers, crucial for initiating amyloidogenesis, remain largely uncharacterized. We report an atomic-resolution structural characterization toxic aggregation intermediate (I1) on...
Abstract We describe a new labeling method that allows for full protonation at the backbone Hα position, maintaining protein side chains with high level of deuteration. refer to as alpha proton exchange by transamination (α-PET) since it relies on transaminase activity demonstrated here using Escherichia coli expression. show α-PET is particularly useful in improving structural characterization solid proteins introduction an additional reporter, while eliminating many strong dipolar...
Homonuclear dipolar recoupling is routinely used for magic-angle spinning NMR-based structure determination. In fully protonated samples, only short proton-proton distances are accessible to broadband approaches because of high proton density. Selective methods allow detection longer by directing polarization a subset spins. Here we introduce the selective pulse sequence MODIST, which recouples spins that have modest chemical shift offset difference, and demonstrate it selectively record...
Structural characterization of protein–ligand binding interfaces at atomic resolution is essential for improving the design specific and potent inhibitors. Herein, we explored fast 19F- 1H-detected magic angle spinning NMR spectroscopy to investigate interaction between two fluorinated ligand diastereomers with microcrystalline galectin-3 carbohydrate recognition domain. The detailed environment around fluorine atoms was mapped by 2D 13C–19F 1H–19F dipolar correlation experiments permitted...
Abstract Determination of the environment surrounding a protein is often key to understanding its function and can also be used infer structural properties protein. By using proton‐detected solid‐state NMR, we show that reduced spin diffusion within under conditions fast magic‐angle spinning, high magnetic field, sample deuteration allows efficient measurement site‐specific exposure mobile water lipids. We demonstrate this site specificity on two membrane proteins, human voltage dependent...
The drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and pore that is primary of efficacy. It remained enigmatic binding did not occur certain detergent micelles, particular incomplete was observed mixture lipids selected match viral membrane. Here we show effects are responsible, namely changes upon prevented solubilization, slow kinetics lipid samples. Using 55-100 kHz magic-angle spinning NMR, characterize three environments: DPhPC, DPhPC...
The flow of ions across cell membranes facilitated by ion channels is an important function for all living cells. Despite the huge amount structural data provided crystallography, elucidating exact interactions between selectivity filter atoms and bound challenging. Here, we detect
Internuclear distance determination is the foundation for NMR-based structure calculation. However, high-precision measurement a laborious process requiring lengthy data acquisitions due to large set of multidimensional spectra needed at different mixing times. This prevents application or challenging molecular systems. Here, we present new approach, transferred-rotational-echo double resonance (TREDOR), heteronuclear transfer method in which simultaneously detect both starting and...
Atomic details of structured water molecules are indispensable to understand the thermodynamics important biological processes including proton conduction mechanism M2 protein. Despite expectation based on crystal structures Influenza A M2, only two populations have been observed by NMR in reconstituted lipid bilayer samples. These bulk- and lipid-associated typically seen membrane Here, we detect a bound molecule at chemical shift 11 ppm, located near functional histidine 37 residue...
Obtaining atomic-level information on components in the cell is a major focus structural biology. Elucidating specific and dynamic features of proteins their interactions cellular context crucial for understanding processes. We introduce 19 F nuclear polarization (DNP) combined with fast magic-angle-spinning (MAS) magnetic resonance (NMR) spectroscopy as powerful technique to study mammalian cells. demonstrate our approach severe acute respiratory syndrome coronavirus 2 5F-Trp-N NTD protein,...
Abstract The drug Rimantadine binds to two different sites in the M2 protein from influenza A, a peripheral site and pore that is primary of efficacy. It remained enigmatic binding did not occur certain detergent micelles, particular incomplete was observed mixture lipids selected match viral membrane. Here we show effects are responsible, namely changes upon prevented solubilization, slow kinetics lipid samples. Using 55-100 kHz magic-angle spinning NMR, characterize three environments:...
The radio-frequency-driven recoupling (RFDR) pulse sequence is used in magic-angle spinning (MAS) NMR to recouple homonuclear dipolar interactions. Here we show simultaneous of both the heteronuclear and interactions by applying RFDR pulses on two channels. We demonstrate method, called HETeronuclear (HET-RFDR), microcrystalline SH3 samples at 10 55.555 kHz MAS. Numerical simulations HET-RFDR standard sequences allow for better understanding influence offsets paths magnetization transfers...
Here we report the structure of Opa60 in lipid bilayers using proton-detected magic-angle spinning nuclear magnetic resonance (MAS NMR). Preparations including near-native oligosaccharide lipids reveal a consistent picture stable transmembrane beta barrel with minor increase structured region as compared previously reported detergent structure. The large variable loops known to interact host proteins could not be detected, confirming their dynamic nature even bilayer environment. provides...
ABSTRACT The arrangement of histidine sidechains in influenza A M2 tetramer determines their pKa values, which define pH controlled proton conduction critical to the virus lifecycle. Both water associated and hydrogen bonded Imidazole–Imidazolium quaternary structures have been proposed, based on crystal structures, NMR chemical shifts, respectively. Here we show, using domain construct lipid bilayers, that imidazole rings are even at a 7.8 neutral charge state. An intermolecular 8.9 ± 0.3...