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Your search keyword '"Penzel, Susanne"' showing total 12 results
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12 results on '"Penzel, Susanne"'

4. Dimer Organization of Membrane-Associated NS5A of Hepatitis C Virus as Determined by Highly Sensitive H-1-Detected Solid-State NMR

Author

Jirasko, Vlastimil, Lends, Alons, Lakomek, Nils-Alexander, Fogeron, Marie-Laure, Weber, Marco E., Malär, Alexander A., Penzel, Susanne, Bartenschlager, Ralf, Meier, Beat H., and Böckmann, Anja

Subjects
cell-free synthesis, hepatitis C virus, NS5A, paramagnetic relaxation enhancement, solid-state NMR, viruses, virus diseases, biochemical phenomena, metabolism, and nutrition, digestive system diseases
Abstract

The Hepatitis C virus nonstructural protein 5A (NS5A) is a membrane-associated protein involved in multiple steps of the viral life cycle. Direct-acting antivirals (DAAs) targeting NS5A are a cornerstone of antiviral therapy, but the mode-of-action of these drugs is poorly understood. This is due to the lack of information on the membrane-bound NS5A structure. Herein, we present the structural model of an NS5A AH-linker-D1 protein reconstituted as proteoliposomes. We use highly sensitive proton-detected solid-state NMR methods suitable to study samples generated through synthetic biology approaches. Spectra analyses disclose that both the AH membrane anchor and the linker are highly flexible. Paramagnetic relaxation enhancements (PRE) reveal that the dimer organization in lipids requires a new type of NS5A self-interaction not reflected in previous crystal structures. In conclusion, we provide the first characterization of NS5A AH-linker-D1 in a lipidic environment shedding light onto the mode-of-action of clinically used NS5A inhibitors. ISSN:1433-7851 ISSN:1521-3773 ISSN:0570-0833

Published
2021

5. 100 kHz MAS Proton-Detected NMR Spectroscopy of Hepatitis B Virus Capsids

Author

Lecoq, Lauriane, Schledorn, Maarten, Wang, Shishan, Smith-Penzel, Susanne, Malär, Alexander, Callon, Morgane, Nassal, Michael, Meier, Beat, Böckmann, Anja, Institut de biologie structurale (IBS - UMR 5075 ), Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry (MMSB), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), University Hospital Freiburg, Physical Chemistry [ETH Zürich], Department of Chemistry and Applied Biosciences [ETH Zürich] (D-CHAB), Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich)- Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
deuteration, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], Brief Research Report, fast MAS, Solid-state NMR, Fast MAS, Proton detection, Carbon detection, Deuteration, Hepatitis B virus, Capsid, Core protein, proton detection, lcsh:Biology (General), [CHIM.ANAL]Chemical Sciences/Analytical chemistry, carbon detection, core protein, capsid, solid-state NMR, Molecular Biosciences, hepatitis B virus, lcsh:QH301-705.5
Abstract

International audience; We sequentially assigned the fully-protonated capsids made from core proteins of the Hepatitis B virus using proton detection at 100 kHz magic-angle spinning (MAS) in 0.7 mm rotors and compare sensitivity and assignment completeness to previously obtained assignments using carbon-detection techniques in 3.2 mm rotors and 17.5 kHz MAS. We show that proton detection shows a global gain of a factor ∼50 in mass sensitivity, but that signal-to-noise ratios and completeness of the assignment was somewhat higher for carbon-detected experiments for comparable experimental times. We also show that deuteration and H N back protonation improves the proton linewidth at 100 kHz MAS by a factor of 1.5, from an average of 170-110 Hz, and by a factor of 1.3 compared to deuterated capsids at 60 kHz MAS in a 1.3 mm rotor. Yet, several H N protons cannot be back-exchanged due to solvent inaccessibility, which results in a total of 15% of the amides missing in the spectra.

Published
2019
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6. Dimer Organization of Membrane‐Associated NS5A of Hepatitis C Virus as Determined by Highly Sensitive 1H‐Detected Solid‐State NMR.

Author

Jirasko, Vlastimil, Lends, Alons, Lakomek, Nils‐Alexander, Fogeron, Marie‐Laure, Weber, Marco E., Malär, Alexander A., Penzel, Susanne, Bartenschlager, Ralf, Meier, Beat H., and Böckmann, Anja

Subjects
HEPATITIS C virus, VIRAL nonstructural proteins, VIRAL proteins, ANTIVIRAL agents, SYNTHETIC biology, STRUCTURAL models, SPECTRUM analysis
Abstract

The Hepatitis C virus nonstructural protein 5A (NS5A) is a membrane‐associated protein involved in multiple steps of the viral life cycle. Direct‐acting antivirals (DAAs) targeting NS5A are a cornerstone of antiviral therapy, but the mode‐of‐action of these drugs is poorly understood. This is due to the lack of information on the membrane‐bound NS5A structure. Herein, we present the structural model of an NS5A AH‐linker‐D1 protein reconstituted as proteoliposomes. We use highly sensitive proton‐detected solid‐state NMR methods suitable to study samples generated through synthetic biology approaches. Spectra analyses disclose that both the AH membrane anchor and the linker are highly flexible. Paramagnetic relaxation enhancements (PRE) reveal that the dimer organization in lipids requires a new type of NS5A self‐interaction not reflected in previous crystal structures. In conclusion, we provide the first characterization of NS5A AH‐linker‐D1 in a lipidic environment shedding light onto the mode‐of‐action of clinically used NS5A inhibitors. [ABSTRACT FROM AUTHOR]

Published
2021
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7. Setting the magic angle for fast magic-angle spinning probes.

Author

Penzel, Susanne, Smith, Albert A., Ernst, Matthias, and Meier, Beat H.

Subjects
*MOLECULAR probes, *MAGIC angle spinning, *CALIBRATION, *GYROMAGNETIC ratio, *MATHEMATICAL optimization
Abstract

Fast magic-angle spinning, coupled with 1 H detection is a powerful method to improve spectral resolution and signal to noise in solid-state NMR spectra. Commercial probes now provide spinning frequencies in excess of 100 kHz. Then, one has sufficient resolution in the 1 H dimension to directly detect protons, which have a gyromagnetic ratio approximately four times larger than 13 C spins. However, the gains in sensitivity can quickly be lost if the rotation angle is not set precisely. The most common method of magic-angle calibration is to optimize the number of rotary echoes, or sideband intensity, observed on a sample of KBr. However, this typically uses relatively low spinning frequencies, where the spinning of fast-MAS probes is often unstable, and detection on the 13 C channel, for which fast-MAS probes are typically not optimized. Therefore, we compare the KBr-based optimization of the magic angle with two alternative approaches: optimization of the splitting observed in 13 C-labeled glycine-ethylester on the carbonyl due to the Cα–C′ J -coupling, or optimization of the H–N J -coupling spin echo in the protein sample itself. The latter method has the particular advantage that no separate sample is necessary for the magic-angle optimization. [ABSTRACT FROM AUTHOR]

Published
2018
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8. 100 kHz MAS Proton-Detected NMR Spectroscopy of Hepatitis B Virus Capsids

Author

Lecoq, Lauriane, Schledorn, Maarten, Wang, Shishan, Smith-Penzel, Susanne, Malär, Alexander A., Callon, Morgane, Nassal, Michael, Meier, Beat H., and Böckmann, Anja

Subjects
Hepatitis B virus, Capsid, Core protein, Deuteration, Fast MAS, Solid-state NMR, 3. Good health, Proton detection, Carbon detection
Abstract

We sequentially assigned the fully-protonated capsids made from core proteins of the Hepatitis B virus using proton detection at 100 kHz magic-angle spinning (MAS) in 0.7 mm rotors and compare sensitivity and assignment completeness to previously obtained assignments using carbon-detection techniques in 3.2 mm rotors and 17.5 kHz MAS. We show that proton detection shows a global gain of a factor ~50 in mass sensitivity, but that signal-to-noise ratios and completeness of the assignment was somewhat higher for carbon-detected experiments for comparable experimental times. We also show that deuteration and HN back protonation improves the proton linewidth at 100 kHz MAS by a factor of 1.5, from an average of 170–110 Hz, and by a factor of 1.3 compared to deuterated capsids at 60 kHz MAS in a 1.3 mm rotor. Yet, several HN protons cannot be back-exchanged due to solvent inaccessibility, which results in a total of 15% of the amides missing in the spectra., Frontiers in Molecular Biosciences, 6, ISSN:2296-889X

10. Dimer Organization of Membrane-Associated NS5A of Hepatitis C Virus as Determined by Highly Sensitive H-1-Detected Solid-State NMR

Author

Jirasko, Vlastimil, Lends, Alons, Lakomek, Nils-Alexander, Fogeron, Marie-Laure, Weber, Marco E., Malär, Alexander A., Penzel, Susanne, Bartenschlager, Ralf, Meier, Beat H., and Böckmann, Anja

Subjects
hepatitis C virus, cell-free synthesis, paramagnetic relaxation enhancement, viruses, virus diseases, solid-state NMR, biochemical phenomena, metabolism, and nutrition, NS5A, digestive system diseases, 3. Good health
Abstract

The Hepatitis C virus nonstructural protein 5A (NS5A) is a membrane-associated protein involved in multiple steps of the viral life cycle. Direct-acting antivirals (DAAs) targeting NS5A are a cornerstone of antiviral therapy, but the mode-of-action of these drugs is poorly understood. This is due to the lack of information on the membrane-bound NS5A structure. Herein, we present the structural model of an NS5A AH-linker-D1 protein reconstituted as proteoliposomes. We use highly sensitive proton-detected solid-state NMR methods suitable to study samples generated through synthetic biology approaches. Spectra analyses disclose that both the AH membrane anchor and the linker are highly flexible. Paramagnetic relaxation enhancements (PRE) reveal that the dimer organization in lipids requires a new type of NS5A self-interaction not reflected in previous crystal structures. In conclusion, we provide the first characterization of NS5A AH-linker-D1 in a lipidic environment shedding light onto the mode-of-action of clinically used NS5A inhibitors., Angewandte Chemie. International Edition, 60 (10), ISSN:1433-7851, ISSN:1521-3773, ISSN:0570-0833

11. Cell-free expression, purification, and membrane reconstitution for NMR studies of the nonstructural protein 4B from hepatitis C virus

Author

Fogeron, Marie-Laure, Jirasko, Vlastimil, Penzel, Susanne, Paul, David, Montserret, Roland, Danis, Clément, Lacabanne, Denis, Badillo, Aurélie, Gouttenoire, Jérôme, Moradpour, Darius, Bartenschlager, Ralf, Penin, François, Meier, Beat H., and Böckmann, Anja

Subjects
Integral membrane protein, Lipid reconstitution, NS4B, Cell-free protein expression, Solid-state NMR, 3. Good health, Isotope labeling
Abstract

Journal of Biomolecular NMR, 65 (2), ISSN:0925-2738, ISSN:1573-5001

12. Efficient low-power TOBSY sequences for fast MAS.

Author

Tan, Kong Ooi, Agarwal, Vipin, Lakomek, Nils-Alexander, Penzel, Susanne, Meier, Beat H., and Ernst, Matthias

Subjects
*NUCLEAR magnetic resonance, *MAGNETIZATION, *PROTONS, *LARMOR frequency, *ZWITTERIONS
Abstract

Through-bond J -coupling based experiments in solid-state NMR spectroscopy are challenging because the J couplings are typically much smaller than the dipolar couplings. This often leads to a lower transfer efficiency compared to dipolar-coupling based sequences. One of the reasons for the low transfer efficiency are the second-order cross terms involving the strong heteronuclear dipolar couplings leading to fast magnetization decay. Here, we show that by employing a symmetry-based C9 sequence, which was carefully selected to suppress second-order terms, efficient polarization transfers of up to 80% can be achieved without decoupling on fully protonated two-spin model systems at a MAS frequency of 55.5 kHz with rf-field amplitudes of about 25 kHz. In addition, we analyse the effects of rf inhomogeneity and crystallites selection due to the polarization preparation method on the TOBSY transfer efficiency. We demonstrate on small model substances as well as on deuterated and 100% back-exchanged ubiquitin that C 9 39 1 and C 9 48 1 are efficient and practical TOBSY sequences at experimental conditions ranging from proton Larmor frequencies of 400–850 MHz, and MAS frequencies ranging from 55.5 to 111.1 kHz. [ABSTRACT FROM AUTHOR]

Published
2018
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