A5044919510- Computational Drug Discovery Methods
- SARS-CoV-2 and COVID-19 Research
- Bacterial Genetics and Biotechnology
- Protein Structure and Dynamics
- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- Cancer-related Molecular Pathways
- Ion Channels and Receptors
- Lipid Membrane Structure and Behavior
- Drug Transport and Resistance Mechanisms
- Calcium signaling and nucleotide metabolism
- Biochemical and Molecular Research
- RNA Research and Splicing
- Legume Nitrogen Fixing Symbiosis
- RNA Interference and Gene Delivery
- ATP Synthase and ATPases Research
- Peptidase Inhibition and Analysis
- interferon and immune responses
- COVID-19 Clinical Research Studies
- Photosynthetic Processes and Mechanisms
- Diverse Scientific Research Studies
- Synthesis and biological activity
- Adenosine and Purinergic Signaling
- Protein Kinase Regulation and GTPase Signaling
Universität Hamburg
2013-2024
Max Planck Institute for the Structure and Dynamics of Matter
2018-2021
Hamburg Centre for Ultrafast Imaging
2019
Institute of Biochemistry
2019
Center for Free-Electron Laser Science
2018
Aarhus University
2010-2013
National Research Foundation
2012
Medical Research Council
2006-2010
Hutchison/MRC Research Centre
2008-2009
University of Cambridge
2006
Proteins with intrinsically disordered domains are implicated in a vast range of biological processes, especially cell signaling and regulation. Having solved the quaternary structure folded tumor suppressor p53 by multidisciplinary approach, we have now determined average ensemble N-terminal transactivation domain (TAD) using residual dipolar couplings (RDCs) from NMR spectroscopy small-angle x-ray scattering (SAXS). Remarkably, not only were able to measure RDCs isolated TAD, but also do...
The coronavirus disease (COVID-19) caused by SARS-CoV-2 is creating tremendous human suffering. To date, no effective drug available to directly treat the disease. In a search for against COVID-19, we have performed high-throughput x-ray crystallographic screen of two repurposing libraries main protease (M
The homotetrameric tumor suppressor p53 consists of folded core and tetramerization domains, linked flanked by intrinsically disordered segments that impede structure analysis x-ray crystallography NMR. Here, we solved the quaternary human in solution a combination small-angle scattering, which defined its shape, NMR, identified domain interfaces showed domains had same intact protein as fragments. We combined data with electron microscopy on immobilized samples provided medium resolution 3D...
Human CA150, a transcriptional activator, binds to and is co-deposited with huntingtin during Huntington's disease. The second WW domain of CA150 three-stranded beta-sheet that folds in vitro microseconds forms amyloid fibers under physiological conditions. We found from exhaustive alanine scanning studies fibrillation this begins its denatured conformations, we identified subset residues critical for fibril formation. used high-resolution magic-angle-spinning NMR on site-specific...
The 25-kDa core domain of the tumor suppressor p53 is inherently unstable and melts at just above body temperature, which makes it susceptible to oncogenic mutations that inactivate by lowering its stability. We determined structure in solution using state-of-the-art isotopic labeling techniques NMR spectroscopy complement crystal structure. was very similar but far more mobile than expected. Importantly, we were able analyze structural environment several buried polar groups, indicated...
Abstract Protein stability in detergent or membrane-like environments is the bottleneck for structural studies on integral membrane proteins (IMP). Irrespective of method to study structure an IMP, solubilization from usually first step workflow. Here, we establish a simple, high-throughput screening identify optimal conditions protein stabilization. We apply differential scanning fluorimetry combination with scattering upon thermal denaturation unfolding proteins. Nine different prokaryotic...
Many microbes and fungi acquire the essential ion Fe3+ through synthesis secretion of high-affinity chelators termed siderophores. In Gram-negative bacteria, these ferric-siderophore complexes are actively taken up using highly specific TonB-dependent transporters (TBDTs) located in outer bacterial membrane (OM). However, detailed mechanism how inner-membrane protein TonB connects to OM as well interplay between siderophore- TonB-binding transporter is still poorly understood. Here, we...
Abstract Differential scanning fluorimetry (DSF) using the inherent fluorescence of proteins (nDSF) is a popular technique to evaluate thermal protein stability in different conditions (e.g. buffer, pH). In many cases, ligand binding increases and often this can be detected as clear shift nDSF experiments. Here, we affinity quantification based on shifts. We present four systems with ligands, ranging from nM high μM. Our study suggests that affinities determined by isothermal analysis are...
Critical assessment of structure prediction (CASP) conducts community experiments to determine the state art in computing protein from amino acid sequence. The process relies on experimental providing information about not yet public or be solved structures, for use as targets. For some targets, is time CASP. Calculated accuracy improved dramatically this round, implying that models should now much more useful resolving many sorts difficulties. To test this, selected seven unsolved targets...
The tumor suppressor p53 consists of four 393-residue chains, each which has two natively unfolded (N- and C-terminal) folded (core tetramerization) domains. Their structural organization is poorly characterized as the protein tends to aggregate, defied crystallization, at limits NMR studies. We first stabilized by mutation make it more suitable for extended study then acquired spectra on full-length various combinations shorter domain constructs. spectrum ( 15 N, 1 H transverse relaxation...
Lysine glutarylation (Kglu) of mitochondrial proteins is associated with glutaryl-CoA dehydrogenase (GCDH) deficiency, which impairs lysine/tryptophan degradation and causes destruction striatal neurons during catabolic crisis subsequent movement disability. By investigating the role Kglu modifications in this disease, we compared brain liver glutarylomes Gcdh-deficient mice. In brain, identified 73 sites on 37 involved various metabolic pathways. Ultrastructural immunogold studies indicated...
Recent structures of full-length ATP-binding cassette (ABC) transporter MsbA in different states indicate large conformational changes during the reaction cycle that involve transient dimerization its nucleotide-binding domains (NBDs). However, a detailed molecular understanding structural and associated kinetics upon ATP binding hydrolysis is still missing. Here, we employed time-resolved small-angle X-ray scattering, initiated by stopped-flow mixing, to investigate accompanying NBD (upon...
Gram-negative bacteria take up the essential ion Fe3+ as ferric-siderophore complexes through their outer membrane using TonB-dependent transporters. However, subsequent route inner differs across many bacterial species and siderophore chemistries is not understood in detail. Here, we report crystal structure of protein FoxB (from Pseudomonas aeruginosa) that involved Fe-siderophore uptake. The revealed a fold with two tightly bound heme molecules. In combination vitro reduction assays vivo...
Plasma-membrane Ca2+-ATPases expel Ca2+ from the cytoplasm and are key regulators of homeostasis in eukaryotes. They autoinhibited under low concentrations. Calmodulin (CaM)-binding to a unique regulatory domain releases autoinhibition activates pump. However, structural basis for this activation, including overall structure calcium pump its complex with calmodulin, is unknown. We previously determined high-resolution calmodulin plasma-membrane Ca2+-ATPase ACA8 revealed bimodular mechanism...
Iron is a key nutrient for almost all living organisms. Paradoxically, it poorly soluble and consequently bioavailable. Bacteria have thus developed multiple strategies to access this metal. One of the most common consists use siderophores, small compounds that chelate ferric iron with very high affinity. Many bacteria are able produce their own siderophores or those produced by other microorganisms (exosiderophores) in piracy strategy. Pseudomonas aeruginosa produces two pyoverdine...
The outer membrane of gram-negative bacteria prevents many antibiotics from reaching intracellular targets. However, some antimicrobials can take advantage iron import transporters to cross this barrier. We showed previously that the thiopeptide antibiotic thiocillin exploits nocardamine xenosiderophore transporter, FoxA, opportunistic pathogen Pseudomonas aeruginosa for uptake. Here, we show FoxA also transports bisucaberin and describe at 2.5 Å resolution crystal structure bound FoxA....
The ATP‐binding cassette transporter MsbA is a lipid flippase, translocating A, glycolipids, and lipopolysaccharides from the inner to outer leaflet of membrane Gram‐negative bacteria. It has been used as model system for time‐resolved structural studies several structures in different states reconstitution systems (detergent/nanodiscs/peptidiscs) are available. However, due limited resolution available structures, detailed information on bound nucleotides remained elusive. Here, we have...
TRPM2 is a Ca 2+ permeable, non-selective cation channel in the plasma membrane that involved innate immune response regulating, for example, chemotaxis neutrophils and cytokine secretion monocytes macrophages. The intracellular adenine nucleotides ADP-ribose (ADPR) 2′-deoxy-ADPR (2dADPR) activate channel, combination with their co-agonist . Interestingly, activation of human (hsTRPM2) by 2dADPR much more effective than ADPR. However, underlying mechanism nucleotides’ differential effect on...
The tumor suppressor p53 is frequently mutated in human cancers. Upon activation it can induce cell cycle arrest or apoptosis. ASPP2 specifically stimulate the apoptotic function of but not arrest, mechanism enhancing pro-apoptotic genes over remains unknown. In this study, we analyzed binding 53BP2 (p53-binding protein 2, C-terminal domain ASPP2) to core and various mutants using biophysical techniques. We found that several mutations (R181E, G245S, R249S, R273H) have different effects on...