Single Channel Recording of the Nuclear Pore Complex

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DESCRIPTION (provided by applicant): The nucleus is the central organelle of eukaryotic cells. The molecular transport between the nucleus and the cytoplasm is mediated only through the nuclear pore complex (NPC), which perforates the double membraned nuclear envelope that defines the nuclear compartment. Our long-term objective is to obtain real-time information about individual channel-gating and protein-translocation events in the single NPC. The information about the nucleocytoplasmic transport through the nanometer-sized protein machine, which does not presently exist, will facilitate our comprehensive understanding of many biological processes and diseases, and will accelerate future pharmaceutical regulation of gene expression. To achieve the goal, we will establish a novel experimental approach for single channel recording of the NPC by improving spatial and time resolutions of scanning electrochemical microscope (SECM) to nanometer and microsecond ranges. While SECM is a well-established method for non-invasive and quantitative measurements of membrane permeability, improvement of the spatial resolution by nanofabricated SECM probes will enable us to measure the single NPC permeability. Real-time information about the individual channel events can be obtained by monitoring their effects on the single NPC permeability with high time resolution. The time-resolved information will allow us to evaluate the transport models whose validity can not be checked by currently-available experimental data, leading to more comprehensive understanding of this important transport process.