Nuclear pore complexes (NPCs) lined with intrinsically disordered FG-domains act as selective gatekeepers for molecular transport between the nucleus and the cytoplasm in eukaryotic cells. The underlying physical mechanism of the intriguing selectivity is still under debate. Here, we probe the transport of ions and transport receptors through biomimetic NPCs consisting of Nsp1 domains attached to the inner surface of solid-state nanopores. We examine both wildtype FG- domains and hydrophilic SG-mutants. FG-nanopores showed a clear selectivity as transport receptors can translocate across the pore whereas other proteins cannot. SG mutant pores lack such selectivity. To unravel this striking difference, we present coarse-grained molecular dynamics simulations that reveal that FG-pores exhibit a high-density, nonuniform protein distribution, in contrast to a uniform and significantly less-dense protein distribution in the SG-mutant. We conclude that the sequence-dependent density distribution of disordered proteins inside the NPC plays a key role for its conductivity and selective permeability.

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The controlled functionalization of surfaces with proteins is crucial for many analytical methods in life science research and biomedical applications. Here, a coating for silica-based surfaces is established which enables stable and selective immobilization of proteins with controlled orientation and tunable surface density. The coating is reusable, retains functionality upon long-term storage in air, and is applicable to surfaces of complex geometry. The protein anchoring method is validated on planar surfaces, and then a method is developed to measure the anchoring process in real time using silicon nitride solid-state nanopores. For surface attachment, polyhistidine tags that are site specifically introduced into recombinant proteins are exploited, and the yeast nucleoporin Nsp1 is used as model protein. Contrary to the commonly used covalent thiol chemistry, the anchoring of proteins via polyhistidine tag is reversible, permitting to take proteins off and replace them by other ones. Such switching in real time in experiments on individual nanopores is monitored using ion conductivity. Finally, it is demonstrated that silica and gold surfaces can be orthogonally functionalized to accommodate polyhistidine-tagged proteins on silica but prevent protein binding to gold, which extends the applicability of this surface functionalization method to even more complex sensor devices.

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We present a coupled simulation approach for fluid–structure–acoustic interactions (FSAI) as an example for strongly surface coupled multi-physics problems. In addition to the multi-physics character, FSAI feature multi-scale properties as a further challenge. In our partitioned approach, the problem is split into spatially separated subdomains interacting via coupling surfaces. Within each subdomain, scalable, single-physics solvers are used to solve the respective equation systems. The surface coupling between them is realized with the scalable open-source coupling tool preCICE described in the “Partitioned Fluid–Structure– Acoustics Interaction on Distributed Data: Coupling via preCICE”. We show how this approach enables the use of existing solvers and present the overall scaling behavior for a three-dimensional test case with a bending tower generating acoustic waves.We run this simulation with different solvers demonstrating the performance of various solvers and the flexibility of the partitioned approach with the coupling tool preCICE. An efficient and scalable in-situ visualization reducing the amount of data in place at the simulation processors before sending them over the network or to a file system completes the simulation environment.@en