Structural Enzymology (Schacherl lab)
The Schacherl lab investigates molecular mechanisms and three-dimensional structures of eukaryotic and prokaryotic proteins and RNA complexes using biophysical methods on different scales.
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Research Group Structural Enzymology
The focus of the Schacherl group is the investigation of structure-function relationships and the dynamics of mainly eukaryotic enzymes and macromolecular machines which perform various functions in the cell and play crucial roles in diverse human diseases.
For this purpose, the group uses various biochemical, biophysical and structural biological methods, such as three-dimensional cryo-electron microscopy and tomography. In addition, the group selects and uses nanobodies to study protein structures and function.
The main protein targets are proteases, kinases and guanylate cyclases. Another focus is on the analysis of macromolecular machines, such as the ribosome. For in vitro studies the group uses single particle analysis cryo-EM. To study the function and regulation of macromolecular machines in situ (inside the cell) as well as their cellular organization, the correlative cryo-light and electron microscopic workflow is used, coupled with sub-tomogram averaging to obtain near-atomic resolution structural information. This also involves working with primary cells and established laboratory cell lines. Here, the group mainly studies neuronal cells.
The Schacherl group is open to collaborations with non-structural biologists who would like to study their target proteins individually at atomic resolution or in a cellular context at near-atomic to nanometer resolution.
A variety of biochemical and biophysical methods are used within the research group. These include the following:
- Cryo-electron microscopy (SPA/single particle analysis)
- Correlative cryo-light and electron microscopic workflow (cryo-CLEM/ET)
- Sub-tomogram averaging (STA)
- Development of specific nanobodies using in vitro selection methods
- Heterologous protein expression in bacteria, insect and human cell lines and their purification
- Isolation of proteins from native tissues
- Site-directed mutagenesis
- Chemical crosslinking
- Surface Plasmon Resonance (SPR)
- Fluorescence spectroscopy and fluorescence anisotropy