A5024016091- Monoclonal and Polyclonal Antibodies Research
- SARS-CoV-2 and COVID-19 Research
- Drug Transport and Resistance Mechanisms
- Heat and Mass Transfer in Porous Media
- Lattice Boltzmann Simulation Studies
- RNA and protein synthesis mechanisms
- Glycosylation and Glycoproteins Research
- Legionella and Acanthamoeba research
- Amino Acid Enzymes and Metabolism
- Advanced biosensing and bioanalysis techniques
- Fluid Dynamics and Turbulent Flows
- Bacteriophages and microbial interactions
- Antibiotic Resistance in Bacteria
- Biotin and Related Studies
- DNA and Nucleic Acid Chemistry
- Metal complexes synthesis and properties
- Virus-based gene therapy research
- Biochemical and Structural Characterization
- Bacterial Genetics and Biotechnology
- Electron Spin Resonance Studies
- Antimicrobial Resistance in Staphylococcus
- Antibiotics Pharmacokinetics and Efficacy
- Innovative Microfluidic and Catalytic Techniques Innovation
- SARS-CoV-2 detection and testing
- Nanocluster Synthesis and Applications
University of Zurich
2016-2023
Linkster Therapeutics (Switzerland)
2021-2023
Roche (Switzerland)
2014-2017
University of Basel
2014
ETH Zurich
2007-2013
Mechanistic and structural studies of membrane proteins require their stabilization in specific conformations. Single domain antibodies are potent reagents for this purpose, but generation relies on immunizations, which impedes selections the presence ligands typically needed to populate defined conformational states. To overcome key limitation, we developed an vitro selection platform based synthetic single named sybodies. target limited hydrophilic surfaces proteins, designed three sybody...
Abstract SARS-CoV-2, the causative agent of COVID-19 1 , features a receptor-binding domain (RBD) for binding to host cell ACE2 protein 1–6 . Neutralizing antibodies that block RBD-ACE2 interaction are candidates development targeted therapeutics 7–17 Llama-derived single-domain (nanobodies, ~15 kDa) offer advantages in bioavailability, amenability, and production storage owing their small sizes high stability. Here, we report rapid selection 99 synthetic nanobodies (sybodies) against RBD by...
Abstract ABC exporters harness the energy of ATP to pump substrates across membranes. Extracellular gate opening and closure are key steps transport cycle, but underlying mechanism is poorly understood. Here, we generated a synthetic single domain antibody (sybody) that recognizes heterodimeric exporter TM287/288 exclusively in presence ATP, which was essential solve 3.2 Å crystal structure outward-facing transporter. The sybody binds an extracellular wing strongly inhibits ATPase activity...
Article7 March 2022Open Access Transparent process Biparatopic sybodies neutralize SARS-CoV-2 variants of concern and mitigate drug resistance Justin D Walter orcid.org/0000-0002-1492-3055 Institute Medical Microbiology, University Zurich, Switzerland Contribution: Conceptualization, Supervision, Investigation, Visualization, Methodology, Writing - original draft, review & editing Search for more papers by this author Melanie Scherer orcid.org/0000-0001-5554-9664 Division Neurological...
ABC exporters pump substrates across the membrane by coupling ATP-driven movements of nucleotide binding domains (NBDs) to transmembrane (TMDs), which switch between inward- and outward-facing (IF, OF) orientations. DEER measurements on heterodimeric exporter TM287/288 from Thermotoga maritima, contains a non-canonical ATP site, revealed that in presence nucleotides transporter exists an IF/OF equilibrium. While was sufficient partially populate OF state, trapping pre- or post-hydrolytic...
The COVID-19 pandemic, caused by the novel coronavirus SARS-CoV-2, has resulted in a global health and economic crisis of unprecedented scale. high transmissibility combined with lack population immunity prevalence severe clinical outcomes, urges rapid development effective therapeutic countermeasures. Here, we report generation synthetic nanobodies, known as sybodies, against receptor-binding domain (RBD) SARS-CoV-2. In an expeditious process taking only twelve working days, sybodies were...
Abstract Legionella pneumophila can cause Legionnaires’ disease and replicates intracellularly in a distinct -containing vacuole (LCV). LCV formation is complex process that involves plethora of type IV-secreted effector proteins. The RidL binds the Vps29 retromer subunit, blocks retrograde vesicle trafficking, promotes intracellular bacterial replication. Here, we reveal 29-kDa N-terminal domain (RidL 2–281 ) adopts “foot-like” fold comprising protruding β-hairpin at its “heel”. deletion...
Nanobodies are emerging tools in a variety of fields such as structural biology, cell imaging, and drug discovery. Here we pioneer the use their spin-labeled variants reporters conformational dynamics membrane proteins using DEER spectroscopy. At example bacterial ABC transporter TM287/288, show that two gadolinium-labeled nanobodies allow us to quantify, via analysis modulation depth traces, fraction transporters adopting outward-facing state under different experimental conditions....
ATP-binding cassette (ABC) transporters are ATP-driven molecular machines, in which ATP binding and hydrolysis the nucleotide-binding domains (NBDs) is chemomechanically coupled to large-scale, alternating access conformational changes transmembrane (TMDs), ultimately leading translocation of substrates across biological membranes. The precise nature structural dynamics behind large-scale transition as well coupling NBD TMD motions still unresolved. In this work, we combine all-atom (MD)...
Phosphocholine molecules decorating bacterial cell wall teichoic acids and outer-membrane lipopolysaccharide have fundamental roles in adhesion to host cells, immune evasion, persistence. Bacteria carrying the operon that performs phosphocholine decoration synthesize after uptake of choline precursor by LicB, a conserved transporter among divergent species. Streptococcus pneumoniae is prominent pathogen where plays role virulence. Here, we present cryo–electron microscopy crystal structures...
Abstract Mycobacterium tuberculosis is protected from antibiotic therapy by a multi-layered hydrophobic cell envelope. Major facilitator superfamily (MFS) transporter Rv1410 and the periplasmic lipoprotein LprG are involved in transport of triacylglycerides (TAGs) that seal mycomembrane. Here, we report 2.7 Å structure mycobacterial homologue, which adopts an outward-facing conformation exhibits unusual transmembrane helix 11 12 extensions protrude ~20 into periplasm. A small, very cavity...
Abstract Green fluorescent proteins (GFPs) are widely used to monitor membrane protein expression, purification, and stability. An ideal reporter should be stable itself provide high sensitivity yield. Here, we demonstrate that a coral ( Galaxea fascicularis ) thermostable GFP (TGP) is by such reasons an improved tag compared the conventional jellyfish GFPs. TGP faithfully reports stability at temperatures near 90 °C (20-min heating). By contrast, limit for two popular GFPs 64 74 °C....
Members of the LRRC8 family form heteromeric assemblies, which function as volume-regulated anion channels. These modular proteins consist a transmembrane pore and cytoplasmic leucine-rich repeat (LRR) domains. Despite their known molecular architecture, mechanism activation role LRR domains in this process has remained elusive. Here we address question by generating synthetic nanobodies, termed sybodies, target domain obligatory subunit LRRC8A. We use these binders to investigate...
ABSTRACT SARS-CoV-2, the causative agent of COVID-19 1 , recognizes host cells by attaching its receptor-binding domain (RBD) to receptor ACE2 2–7 . Neutralizing antibodies that block RBD-ACE2 interaction have been a major focus for therapeutic development 8–18 Llama-derived single-domain (nanobodies, ∼15 kDa) offer advantages including ease production and possibility direct delivery lungs nebulization 19 which are attractive features bio-drugs against global respiratory disease. Here, we...