A5053498446- Enzyme Structure and Function
- Advanced Electron Microscopy Techniques and Applications
- Protein Structure and Dynamics
- Ion Channels and Receptors
- Force Microscopy Techniques and Applications
- Photosynthetic Processes and Mechanisms
- RNA and protein synthesis mechanisms
- Microtubule and mitosis dynamics
- Cellular Mechanics and Interactions
- Ion channel regulation and function
- Bacterial Genetics and Biotechnology
- Advanced Fluorescence Microscopy Techniques
- Pancreatic function and diabetes
- Electron and X-Ray Spectroscopy Techniques
- Plant Stress Responses and Tolerance
- Monoclonal and Polyclonal Antibodies Research
- Metabolism, Diabetes, and Cancer
- Bacteriophages and microbial interactions
- Cardiomyopathy and Myosin Studies
- Heat shock proteins research
- Protein Kinase Regulation and GTPase Signaling
- PARP inhibition in cancer therapy
- Skin and Cellular Biology Research
- Lipid Membrane Structure and Behavior
- Endodontics and Root Canal Treatments
Max Planck Institute for Biology
2022-2024
Max Planck Institute for Developmental Biology
2020-2024
UCB Pharma (Germany)
2023
Max Planck Institute of Molecular Physiology
2016-2022
University of Chile
2008-2018
Max Planck Institute for Molecular Biomedicine
2013-2015
University of Münster
2014-2015
Abstract Selecting particles from digital micrographs is an essential step in single-particle electron cryomicroscopy (cryo-EM). As manual selection of complete datasets—typically comprising thousands particles—is a tedious and time-consuming process, numerous automatic particle pickers have been developed. However, non-ideal datasets pose challenge to picking. Here we present the picking software crYOLO which based on deep-learning object detection system You Only Look Once (YOLO). After...
SPHIRE (SPARX for High-Resolution Electron Microscopy) is a novel open-source, user-friendly software suite the semi-automated processing of single particle electron cryo-microscopy (cryo-EM) data. The protocol presented here describes in detail how to obtain near-atomic resolution structure starting from cryo-EM micrograph movies by guiding users through all steps determination pipeline. These are controlled new graphical user interface and require minimum intervention. Using this protocol,...
Canonical transient receptor channels (TRPC) are non-selective cation channels. They involved in receptor-operated Ca2+ signaling and have been proposed to act as store-operated (SOC). Their malfunction is related cardiomyopathies their modulation by small molecules has shown be effective against renal cancer cells. The molecular mechanism underlying the complex activation regulation poorly understood. Here, we report electron cryo-microscopy structure of zebrafish TRPC4 its unliganded...
The actin cytoskeleton drives many essential biological processes, from cell morphogenesis to motility. Assembly of functional networks requires control over the speed at which filaments grow. How this can be achieved high and variable levels soluble subunits found in cells is unclear. Here we reconstitute assembly mammalian, non-muscle physiological concentrations profilin-actin. We discover that under these conditions, filament growth limited by profilin dissociating end elongation becomes...
Abstract Heterodimeric capping protein (CP/CapZ) is an essential factor for the assembly of branched actin networks, which push against cellular membranes to drive a large variety processes. Aside from terminating filament growth, CP potentiates nucleation filaments by Arp2/3 complex in networks through unclear mechanism. Here, we combine structural biology with vitro reconstitution demonstrate that not only terminates elongation, but indirectly stimulates activity activating promoting...
Proteins are the essential agents of all living systems. Even though they synthesized as linear chains amino acids, must assume specific three-dimensional structures in order to manifest their biological activity. These fully specified acid sequences - and therefore nucleotide genes. However, relationship between sequence structure, known protein folding problem, has remained elusive for half a century, despite sustained efforts. To measure progress on this series doubly blind, biennial...
Lifeact is a short actin-binding peptide that used to visualize filamentous actin (F-actin) structures in live eukaryotic cells using fluorescence microscopy. However, this popular probe has been shown alter cellular morphology by affecting the structure of cytoskeleton. The molecular basis for such artefacts poorly understood. Here, we determined high-resolution Lifeact–F-actin complex electron cryo-microscopy (cryo-EM). reveals interacts with hydrophobic binding pocket on F-actin and...
Canonical transient receptor potential channels (TRPC) are involved in receptor-operated and/or store-operated Ca2+ signaling. Inhibition of TRPCs by small molecules was shown to be promising treating renal diseases. In cells, the regulated calmodulin (CaM). Molecular details both CaM and drug binding have remained elusive so far. Here, we report structures TRPC4 complex with three pyridazinone-based inhibitors CaM. The reveal that all bind same cavity voltage-sensing-like domain allow us...
In metazoans, a ≈1 megadalton (MDa) multiprotein complex comprising the dynein-dynactin adaptor Spindly and ROD-Zwilch-ZW10 (RZZ) is building block of fibrous biopolymer, kinetochore corona. The corona assembles on mitotic kinetochores to promote microtubule capture spindle assembly checkpoint (SAC) signaling. We report here high-resolution cryo-EM structure that captures essential features RZZ complex, including farnesyl-binding site required for binding. Using highly predictive in vitro...
SPHIRE (SPARX for High-Resolution Electron Microscopy) is a novel open-source, user-friendly software suite the semi-automated processing of single particle electron cryo-microscopy (cryo-EM) data. The protocol presented here describes in detail how to obtain near-atomic resolution structure starting from cryo-EM micrograph movies by guiding users through all steps determination pipeline. These are controlled new graphical user interface and require minimum intervention. Using this protocol,...
Highly specific transcriptional regulation depends on the cooperative association of transcription factors into enhanceosomes. Usually, their DNA-binding cooperativity originates from either direct interactions or DNA-mediated allostery. Here, we performed unbiased molecular simulations followed by protein-DNA unbinding and free energy profiling to study DNA recognition OCT4 SOX2, key components enhanceosomes in pluripotent cells. We found that SOX2 influences orientation dynamics DNA-bound...
Abstract Bacterial human pathogens secrete initially inactive nucleotidyl cyclases that become potent enzymes by binding to actin inside eukaryotic host cells. The underlying molecular mechanism of this activation is, however, unclear. Here, we report structures ExoY from Pseudomonas aeruginosa and Vibrio vulnificus bound their corresponding activators F-actin profilin-G-actin. reveal in contrast the apo-state, two flexible regions ordered interact strongly with actin. specific stabilization...
Some hyperthermophilic archaea use a modified glycolytic pathway that employs an ADP-dependent glucokinase (ADP-GK) and phosphofructokinase (ADP-PFK) or, in the case of Methanococcus jannaschii, bifunctional glucophosphofructokinase (ADP-GK/PFK). The crystal structures three ADP-GKs have been determined. However, there is no structural information available for ADP-PFKs or ADP-GK/PFK. Here, we present first structure ADP-PFK from Pyrococcus horikoshii OT3 (PhPFK) both apo- AMP-bound forms...
One central goal in molecular evolution is to pinpoint the mechanisms and evolutionary forces that cause an enzyme change its substrate specificity; however, these processes remain largely unexplored. Using glycolytic ADP-dependent kinases of archaea, including orders Thermococcales, Methanosarcinales, Methanococcales, as a model employing approach involving paleoenzymology, statistics, protein structural analysis, we could track changes specificity during kinase along with determinants...