Direct Visualization of Single Nuclear Pore Complex Proteins Using Genetically‐Encoded Probes for DNA‐PAINT. (21st August 2019)

• Record Type: Journal Article
• Authors: 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
• Is Part Of: Angewandte Chemie. Volume 131:Number 37(2019)
• Page Start: 13138
• Page End: 13142
• Publication Date: 2019-08-21
• Subjects: DNA-PAINT -- Einzelmolekül-Bildgebung -- Genetisch kodierte Marker -- Kernporenkomplex -- Superauflösende Mikroskopie; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201905685
• Languages: English
• ISSNs: 0044-8249

Abstract

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. Abstract : Tiefer Einblick in die Zelle : Genetisch kodierte selbstmarkierende Enzyme wie z. B. SNAP‐tag und HaloTag werden mit DNA‐PAINT‐Mikroskopie kombiniert. Dies ermöglicht die Untersuchung von Multikomponentenkomplexen in Zellen auf dem Niveau einzelner Proteine.