Edward Lemke: Decoding Molecular Plasticity in the Dark Proteome
Speaker: Edward Lemke, Departments of Biology and Chemistry, Biocentre Johannes Gutenberg-University Mainz
Chair: Claus Seidel, Institut für Physikalische Chemie, Heinrich-Heine-Universität Düsseldorf
Organizers: Study Group Biophysical Chemistry
The mechanisms by which intrinsically disordered proteins (IDPs) engage in rapid and highly selective binding is a subject of considerable interest and represents a central paradigm to nuclear pore complex (NPC) function, where nuclear transport receptors (NTRs) move through the NPC by binding disordered phenylalanine-glycine-rich nucleoporins (FG-Nups). In the first part of my talk, I will present a combined fluorescence and microfluidics approach that provides a simple model system to study how selective transport in the living cell might be orchestrated. Since site-specific labeling of proteins with small but highly photostable fluorescent dyes inside cells remains the major bottleneck for directly study protein dynamics in the interior of the cell, I will demonstrate an approach how to overcome this limitation in the second part of my talk. We have now developed a semi-synthetic strategy based on novel artificial amino acids that are easily and site-specifically introduced into any protein by the natural machinery of the living cell via a newly developed synthetic organelle which equips the living cell with two genetic codes.. This allows rapid, specific “click” labeling and even multi-color studies of living cells and subsequent super resolution microscopy.
Celetti G, Paci G, Caria J, VanDelinder V, Bachand G, Lemke EA. The liquid state of FG-nucleoporins mimics permeability barrier properties of nuclear pore complexes. J Cell Bio. (2020) Jan 6;219(1). pii: e201907157
Reinkemeier CD, Estrada Girona G, Lemke EA, 2019 Designer membraneless organelles en-able codon reassignment of selected mRNAs in eukaryotes. Science, Mar 29;363(6434)
Nikić I, Estrada Girona G, Kang JH, Paci G, Mikhaleva S, Koehler C, Shymanska NV, Ventura Santos C, Spitz D, Lemke EA*. 2016. Debugging eukaryotic genetic code expansion for site-specific click-PAINT super-resolution microscopy. Angew Chem Int Ed Engl. 55(52):16172-16176