title: Single-point single-molecule FRAP distinguishes inner and outer nuclear membrane protein distribution.
abstract: The normal distribution of nuclear envelope transmembrane proteins (NETs) is disrupted in several human diseases. NETs are synthesized on the endoplasmic reticulum and then transported from the outer nuclear membrane (ONM) to the inner nuclear membrane (INM). Quantitative determination of the distribution of NETs on the ONM and INM is limited in available approaches, which moreover provide no information about translocation rates in the two membranes. Here we demonstrate a single-point single-molecule FRAP microscopy technique that enables determination of distribution and translocation rates for NETs in vivo.
authors: Department of Biology, Temple University, Philadelphia, Pennsylvania 19122, USA.
authors: Krishna C
authors: http://orcid.org/0000-0002-5991-9375
authors: KC
authors: Mudumbi
authors: The Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3BF, UK.
authors: Eric C
authors: EC
authors: Schirmer
authors: Department of Biology, Temple University, Philadelphia, Pennsylvania 19122, USA.
authors: Weidong
authors: W
authors: Yang
mesh_terms: D001692:Biological Transport; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005240:Feasibility Studies; D036681:Fluorescence Recovery After Photobleaching / Q000295:instrumentation / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D009685:Nuclear Envelope / Q000378:metabolism*; D000072760:Single Molecule Imaging / Q000295:instrumentation / Q000379:methods*
chemical_list: D008565:Membrane Proteins