Open AccessDetermination of Membrane Protein Distribution on the Nuclear Envelope by Single‐Point Single‐Molecule FRAP
Author(s) -
Mudumbi Krishna C.,
Yang Weidong
Publication year - 2018
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/cpcb.27
Subject(s) - fluorescence recovery after photobleaching , endoplasmic reticulum , photobleaching , biophysics , membrane , inner membrane , envelope (radar) , transmembrane protein , chemistry , chromosomal translocation , distribution (mathematics) , green fluorescent protein , membrane protein , microbiology and biotechnology , fluorescence , biology , biochemistry , physics , computer science , gene , telecommunications , mathematical analysis , radar , receptor , mathematics , quantum mechanics
Nuclear envelope transmembrane proteins (NETs) are synthesized on the endoplasmic reticulum and then transported from the outer nuclear membrane (ONM) to the inner nuclear membrane (INM) in eukaryotic cells. The abnormal distribution of NETs has been associated with many human diseases. However, quantitative determination of the spatial distribution and translocation dynamics of NETs on the ONM and INM is still very limited in currently existing approaches. Here we demonstrate a single‐point single‐molecule fluorescence recovery after photobleaching (FRAP) microscopy technique that enables quick determination of distribution and translocation rates for NETs in vivo . © 2017 by John Wiley & Sons, Inc.