SKM 2023 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 26: Focus Session mRNA Physics

BP 26.1: Hauptvortrag

Donnerstag, 30. März 2023, 15:00–15:30, TOE 317

Decoding Molecular Plasticity in the Dark Proteome of the Nuclear Transport Machinery — •Edward Lemke — JGU & IMB Mainz, GE

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. Nuclear transport receptors (NTRs) can move small proteins and mRNA through the central channel of the NPC which is filled with hundreds of phenylalanine-glycine-rich nucleoporins (FG-Nups) reaching millimolar concentrations with elusive conformational plasticity. We have now developed a semi-synthetic strategy to equip the living cell with up to three genetic codes and label FG-Nups inside functional NPCs site-specifically with small FRET probes. This allowed us to develop an experimental approach and use fluorescent lifetime imaging microscopy (FLIM) to directly decipher the plasticity of FG-Nups. Our study enabled a conformational look on the densely packed IDPs in the sub-resolution (roughly (50 nm)^3 small cavity) cavity of the NPC. We extracted the scaling exponent, which directly describes the conformations of FG-Nups at their functional status as well as the solvent quality in the cellular and even inner NPC environment. Pairing our data with coarse grained simulations enabled us to complement the missing half of protein mass that due to its dynamics are not present in even the most recent electron tomograms of the NPC.