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Direct Visualization of Single Nuclear Pore Complex Proteins Using Genetically‐Encoded Probes for DNA‐PAINT
Author(s) -
Schlichthaerle Thomas,
Strauss Maximilian T.,
Schueder Florian,
Auer Alexander,
Nijmeijer Bianca,
Kueblbeck Moritz,
Jimenez Sabinina Vilma,
Thevathasan Jervis V.,
Ries Jonas,
Ellenberg Jan,
Jungmann Ralf
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201905685
Subject(s) - nuclear pore , nucleoporin , dna , fluorescence microscope , microbiology and biotechnology , nuclear transport , microscopy , computational biology , biophysics , resolution (logic) , chemistry , nanotechnology , electron microscope , biology , fluorescence , cell nucleus , cytoplasm , materials science , biochemistry , physics , computer science , optics , artificial intelligence
Abstract The nuclear pore complex (NPC) is one of the largest and most complex protein assemblies in the cell and, among other functions, serves as the gatekeeper of nucleocytoplasmic transport. Unraveling its molecular architecture and functioning has been an active research topic for decades with recent cryogenic electron microscopy and super‐resolution studies advancing our understanding of the architecture of the NPC complex. However, the specific and direct visualization of single copies of NPC proteins is thus far elusive. Herein, we combine genetically‐encoded self‐labeling enzymes such as SNAP‐tag and HaloTag with DNA‐PAINT microscopy. We resolve single copies of nucleoporins in the human Y‐complex in three dimensions with a precision of circa 3 nm, enabling studies of multicomponent complexes on the level of single proteins in cells using optical fluorescence microscopy.