Combined Use of Unnatural Amino Acids Enables Dual Color Super-Resolution Imaging of Proteins via Click Chemistry

2018 | journal article; research paper. A publication with affiliation to the University of Göttingen.

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​Saal, Kim-A., Frank Richter, Peter Rehling, and Silvio O. Rizzoli. "Combined Use of Unnatural Amino Acids Enables Dual Color Super-Resolution Imaging of Proteins via Click Chemistry​." ​ACS Nano ​2018, no. 12 (2018): ​12247​-12254​. ​https://doi.org/10.1021/acsnano.8b06047.

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Authors
Saal, Kim-A.; Richter, Frank; Rehling, Peter ; Rizzoli, Silvio O. 
Abstract
Recent advances in optical nanoscopy have brought the imaging resolution to the size of the individual macromolecules, thereby setting stringent requirements for the fluorescent labels. Such requirements are optimally fulfilled by the incorporation of unnatural amino acids (UAAs) in the proteins of interest (POI), followed by fluorophore conjugation via click chemistry. However, this approach has been limited to single POIs in mammalian cells. Here we solve this problem by incorporating different UAAs in different POIs, which are expressed in independent cell sets. The cells are then fused, thereby combining the different proteins and organelles, and are easily imaged by dual-color super-resolution microscopy. This procedure, which we termed Fuse2Click, is simple, requires only the well-established Amber codon, and allows the use of all previously optimized UAAs and tRNA/RS pairs. This should render it a tool of choice for multi-color click-based imaging.
Issue Date
2018
Journal
ACS Nano 
Project
info:eu-repo/grantAgreement/EC/FP7/614765/EU//NEUROMOLANATOMY
info:eu-repo/grantAgreement/EC/FP7/339580/EU//MITRAC
SFB 1190: Transportmaschinen und Kontaktstellen zellulärer Kompartimente 
SFB 1190 | P09: Proteinsortierung in der Synapse: Prinzipien und molekulare Organisation 
SFB 1286: Quantitative Synaptologie 
SFB 1286 | A06: Mitochondrienfunktion und -umsatz in Synapsen 
Organization
Institut für Neuro- und Sinnesphysiologie ; DFG Forschungszentrum Molekularphysiologie des Gehirns und Exzellenzcluster Mikroskopie im Nanometerbereich ; Institut für Zellbiochemie ; Center for Biostructural Imaging of Neurodegeneration 
Working Group
RG Rehling (Mitochondrial Protein Biogenesis) 
RG Rizzoli (Quantitative Synaptology in Space and Time) 
External URL
https://sfb1190.med.uni-goettingen.de/production/literature/publications/47
https://sfb1286.uni-goettingen.de/literature/publications/2
ISSN
1936-086X
Language
English

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