Tobias Schubeis

Tobias Schubeis
Ecole normale supérieure de Lyon | ENS Lyon · CRMN

PhD

About

29
Publications
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Publications

Publications (29)
Article
A new heteronuclear decoupling pulse sequence is introduced, dubbed ROtor-Synchronized Phase-Alternated Cycles (ROSPAC). It is based on a partial refocusing of the coherences (spin operator products, or cross-terms) 1,2 responsible for transverse spin-polarization dephasing, on the irradiation of a large pattern of radio-frequencies, and on a signi...
Article
Since the first pioneering studies on small deuterated peptides dating more than 20 years ago, 1H detection has evolved into the most efficient approach for investigation of biomolecular structure, dynamics, and interactions by solid-state NMR. The development of faster and faster magic-angle spinning (MAS) rates (up to 150 kHz today) at ultrahigh...
Article
Full-text available
The CorA family of proteins regulates the homeostasis of divalent metal ions in many bacteria, archaea, and eukaryotic mitochondria, making it an important target in the investigation of the mechanisms of transport and its functional regulation. Although numerous structures of open and closed channels are now available for the CorA family, the mech...
Article
Full-text available
The E.coli maltose binding protein (MBP) is a 42.5 kDa molecule widely employed in many biotechnology applications. Because of its molecular size, it has become the main model system for the development of solution NMR methods adapted to large biomolecular targets. Here, we report virtually complete (~ 90%) backbone resonance assignments obtained o...
Preprint
Full-text available
The CorA family of proteins regulates the homeostasis of divalent metal ions in many bacteria, archaea, and eukaryotic mitochondria, making it an important target in the investigation of the mechanisms of transport and its functional regulation. Although numerous structures of open and closed channels are now available for the CorA family, the mech...
Article
Structure determination of adjuvant‐coupled antigens is essential for rational vaccine development but has so far been hampered by the relatively low antigen content in vaccine formulations and by their heterogeneous composition. Here we show that magic‐angle spinning (MAS) solid‐state NMR can be used to assess the structure of the influenza virus...
Article
Structure determination of adjuvant‐coupled antigens is essential for rational vaccine development but has so far been hampered by the relatively low antigen content in vaccine formulations and by their heterogeneous composition. Here we show that magic‐angle spinning (MAS) solid‐state NMR can be used to assess the structure of the influenza virus...
Article
Copper/zinc superoxide dismutase (SOD) is a homodimeric metalloenzyme that has been extensively studied as a benchmark for structure-function relationships in proteins, in particular because of its implication in the familial form of the neurodegenerative disease amyotrophic lateral sclerosis. Here, we investigate microcrystalline preparations of t...
Article
Full-text available
Most commonly small outer membrane proteins, possessing between 8 and 12 β-strands, are not involved in transport but fulfill diverse functions such as cell adhesion or binding of ligands. An intriguing exception are the 8-stranded β-barrel proteins of the OmpW family, which are implicated in the transport of small molecules. A representative examp...
Article
Full-text available
The protein AlkL is known to increase permeability of the outer membrane of bacteria for hydrophobic molecules, yet the mechanism of transport has not been determined. Differing crystal and NMR structures of homologous proteins resulted in a controversy regarding the degree of structure and the role of long extracellular loops. Here we solve this c...
Article
Thanks to magic-angle spinning (MAS) probes with frequencies of 60–100 kHz, the benefit of high sensitivity 1H detection can now be broadly realized in biomolecular solid-state NMR for the analysis of microcrystalline, sedimented, or lipid-embedded preparations. Nonetheless, performing the assignment of all resonances remains a rate-limiting step i...
Article
Protein‐Spins in einer neuen Dimension: Korrelationen von fünf Resonanzfrequenzen von Kernspins innerhalb eines NMR‐Experiments sind jetzt für schnell rotierende Protein‐Festkörper mithilfe der Projektionsspektroskopie möglich. Aufgrund der drastisch verringerten Mehrdeutigkeit kann die Proteinrückgrat‐Resonanzzuordnung automatisch unter Umgehung d...
Article
Narrow proton signals, high sensitivity, and efficient coherence transfers provided by fast magic‐angle spinning at high magnetic fields make automated projection spectroscopy feasible in protein solid‐state NMR. We present the first ultra‐high dimensional implementation of this approach where 5D peak lists are reconstructed from a number of 2D pro...
Article
Determination of the environment surrounding a protein is often key to understanding function, and can also be used to infer structural properties of the protein itself. Using proton‐detected solid‐state NMR, we show that reduced spin diffusion within the protein under conditions of fast magic‐angle spinning, high magnetic field, and sample deutera...
Article
Curli are functional amyloids that form a major part of the biofilm produced by many enterobacteriaceae. A multi protein system around the outer membrane protein CsgG is in charge of the export and controlled propagation of the main Curli subunits, CsgA and CsgB. CsgF is essential for the linkage of the main amyloid-forming proteins to the cell sur...
Article
Building on a decade of continuous advances of the community, the recent development of very fast (60 kHz and above) magic-angle spinning (MAS) probes has revolutionised the field of solid-state NMR. This new spinning regime reduces the ¹H-¹H dipolar couplings, so that direct detection of the larger magnetic moment available from ¹H is now possible...
Chapter
Solid-state NMR spectroscopy (ssNMR) is uniquely suited for atomic-resolution structural investigations of large protein assemblies, which are notoriously difficult to study due to their insoluble and non- crystalline nature. However, assignment ambiguities because of limited resolution and spectral crowding are currently major hurdles that quickly...
Article
We present the first solid-state NMR study of an iron-sulfur protein. The combined use of very fast (60 kHz) magic-angle spinning and tailored radiofrequency irradiation schemes allows the detection and the assignment of most of the (1)H and (13)C resonances of the oxidized high-potential iron-sulfur protein I from Ectothiorhodospira halophila (EhH...
Article
Very fast magic-angle spinning (MAS>80 kHz) NMR spectroscopy combined with high field magnets has enabled the acquisition of proton-detected spectra in fully protonated solid samples with sufficient resolution and sensitivity. One of the primary challenges in structure determination of protein is to observe long-range 1H-1H proximities. We demonstr...
Article
Protein Structures G. Pintacuda et al. show in their Communication on page 15504 ff. that with 111 kHz magic-angle spinning probes, high-resolution ¹H-detected solid-state NMR spectra of proteins can be acquired without deuteration.
Article
Festkörper-NMR-Spektroskopie Neue MAS-Sonden ermöglichen die hochauflösende NMR-Spektroskopie von unlöslichen, kristallinen oder selbstorganisierten Proteinaggregaten ohne Deuterierung. G. Pintacuda et al. beschreiben die Methode in der Zuschrift auf S. 15730 ff.
Article
Wir beschreiben die empfindliche Messung von α-Protonen in vollständig protonierten Proteinen durch Festkörper-NMR-Spektroskopie mit Rotation um den magischen Winkel (MAS) von 100–111 kHz. Die exzellente Auflösung in der Cα-Hα-Ebene wird für fünf Proteine gezeigt, darunter Mikrokristalle, ein sedimentierter Komplex, ein Kapsid und Amyloidfibrillen....
Article
We demonstrate sensitive detection of alpha protons of fully protonated proteins by solid-state NMR spectroscopy with 100–111 kHz magic-angle spinning (MAS). The excellent resolution in the Cα-Hα plane is demonstrated for 5 proteins, including microcrystals, a sedimented complex, a capsid and amyloid fibrils. A set of 3D spectra based on a Cα–Hα de...
Article
Dynamic nuclear polarization (DNP) NMR can enhance sensitivity but often comes at the price of a substantial loss of resolution. Two major factors affect spectral quality, low temperature heterogeneous line broadening and paramagnetic relaxation enhancement (PRE) effects. Investigations by NMR, isothermal titration calorimetry (ITC) and electron pa...
Article
Curli are functional bacterial amyloids produced by an intricate biogenesis machinery. Insights into their folding and regulation can advance our understanding of amyloidogenesis. However, gaining detailed structural information of amyloids, and their tendency for structural polymorphisms, remains challenging. Herein we compare high-quality solid-s...
Article
Curli sind funktionelle bakterielle Amyloide, die durch eine ausgefeilte Biogenesemaschinerie hergestellt werden. Einblicke in ihre Faltung und Regulierung kann unser Verständnis der Amyloidbildung erweitern. Allerdings ist die Aufklärung hochaufgelöster Strukturen von Amyloiden aufgrund ihrer Tendenz zu strukturellen Polymorphismen weiterhin eine...
Article
Resolution and sensitivity in solid state NMR (SSNMR) can rival the results achieved by solution NMR, and even outperform them in the case of large systems. However, several factors affect the spectral quality in SSNMR samples, and not all systems turn out to be equally amenable for this methodology. In this review we attempt at analyzing the cause...
Article
We present an efficient method for the reduction of spectral complexity in the solid-state NMR spectra of insoluble protein assemblies, without loss of signal intensity. The approach is based on segmental isotope labeling by using the split intein DnaE from Nostoc punctiforme. We show that the segmentally (13) C,(15) N-labeled prion domain of HET-s...