Bioorthogonal Click Chemistry Enables Site-specific Fluorescence Labeling of Functional NMDA Receptors for Super-Resolution Imaging

2018 ◽  
Vol 57 (50) ◽  
pp. 16364-16369 ◽  
Author(s):  
Franziska Neubert ◽  
Gerti Beliu ◽  
Ulrich Terpitz ◽  
Christian Werner ◽  
Christian Geis ◽  
...  
Keyword(s):  
Nmda Receptors ◽  
Click Chemistry ◽  
Super Resolution ◽  
Fluorescence Labeling ◽  
Site Specific ◽  
Specific Fluorescence ◽  
Resolution Imaging

Bioorthogonal Click Chemistry Enables Site-specific Fluorescence Labeling of Functional NMDA Receptors for Super-Resolution Imaging

2018 ◽  
Vol 130 (50) ◽  
pp. 16602-16607 ◽  
Author(s):  
Franziska Neubert ◽  
Gerti Beliu ◽  
Ulrich Terpitz ◽  
Christian Werner ◽  
Christian Geis ◽  
...  
Keyword(s):  
Nmda Receptors ◽  
Click Chemistry ◽  
Super Resolution ◽  
Fluorescence Labeling ◽  
Site Specific ◽  
Specific Fluorescence ◽  
Resolution Imaging

scholarly journals Genetic Code Expansion and Click-Chemistry Labeling to Visualize GABA-A Receptors by Super-Resolution Microscopy

2021 ◽  
Vol 13 ◽  
Author(s):  
Alexander Kuhlemann ◽  
Gerti Beliu ◽  
Dieter Janzen ◽  
Enrica Maria Petrini ◽  
Danush Taban ◽  
...  
Keyword(s):  
Genetic Code ◽  
Click Chemistry ◽  
Unnatural Amino Acids ◽  
Super Resolution ◽  
Receptor Expression ◽  
Fluorescence Labeling ◽  
Site Specific ◽  
Gaba A ◽  
Genetic Code Expansion ◽  
Super Resolution Microscopy

Fluorescence labeling of difficult to access protein sites, e.g., in confined compartments, requires small fluorescent labels that can be covalently tethered at well-defined positions with high efficiency. Here, we report site-specific labeling of the extracellular domain of γ-aminobutyric acid type A (GABA-A) receptor subunits by genetic code expansion (GCE) with unnatural amino acids (ncAA) combined with bioorthogonal click-chemistry labeling with tetrazine dyes in HEK-293-T cells and primary cultured neurons. After optimization of GABA-A receptor expression and labeling efficiency, most effective variants were selected for super-resolution microscopy and functionality testing by whole-cell patch clamp. Our results show that GCE with ncAA and bioorthogonal click labeling with small tetrazine dyes represents a versatile method for highly efficient site-specific fluorescence labeling of proteins in a crowded environment, e.g., extracellular protein domains in confined compartments such as the synaptic cleft.

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

ACS Nano ◽  
2018 ◽  
Vol 12 (12) ◽  
pp. 12247-12254 ◽  
Author(s):  
Kim-A. Saal ◽  
Frank Richter ◽  
Peter Rehling ◽  
Silvio O. Rizzoli
Keyword(s):  
Amino Acids ◽  
Click Chemistry ◽  
Unnatural Amino Acids ◽  
Super Resolution ◽  
Dual Color ◽  
Combined Use ◽  
Resolution Imaging

scholarly journals Super-Resolution Microscopy of Chromatin

Genes ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 493 ◽  
Author(s):  
Birk
Keyword(s):  
Gene Regulation ◽  
Data Analysis ◽  
Super Resolution ◽  
Fluorescence Labeling ◽  
Cellular Processes ◽  
Direct Assessment ◽  
Chromatin Interactions ◽  
Resolution Imaging ◽  
Super Resolution Microscopy ◽  
Insight Into

Since the advent of super-resolution microscopy, countless approaches and studies have been published contributing significantly to our understanding of cellular processes. With the aid of chromatin-specific fluorescence labeling techniques, we are gaining increasing insight into gene regulation and chromatin organization. Combined with super-resolution imaging and data analysis, these labeling techniques enable direct assessment not only of chromatin interactions but also of the function of specific chromatin conformational states.

scholarly journals Bioorthogonal double-fluorogenic siliconrhodamine probes for intracellular super-resolution microscopy

2017 ◽  
Vol 53 (50) ◽  
pp. 6696-6699 ◽  
Author(s):  
E. Kozma ◽  
G. Estrada Girona ◽  
G. Paci ◽  
E. A. Lemke ◽  
P. Kele
Keyword(s):  
Super Resolution ◽  
Site Specific ◽  
Intracellular Proteins ◽  
Resolution Imaging ◽  
Super Resolution Microscopy

A series of double-fluorogenic siliconrhodamine-tetrazines were synthesized. One of these tetrazines is a membrane-permeant label allowing site-specific bioorthogonal tagging of intracellular proteins and super-resolution imaging.

Mapping Calcium-Sensitive Regions in the Neuronal Calcium Sensor GCAP2 by Site-Specific Fluorescence Labeling

Biochemistry ◽  
2016 ◽  
Vol 55 (18) ◽  
pp. 2567-2577 ◽  
Author(s):  
Stefan Sulmann ◽  
Melanie Wallisch ◽  
Alexander Scholten ◽  
Jens Christoffers ◽  
Karl-Wilhelm Koch
Keyword(s):  
Fluorescence Labeling ◽  
Calcium Sensor ◽  
Site Specific ◽  
Specific Fluorescence ◽  
Neuronal Calcium Sensor

scholarly journals Correction: Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Tino Pleiner ◽  
Mark Bates ◽  
Sergei Trakhanov ◽  
Chung-Tien Lee ◽  
Jan Erik Schliep ◽  
...  
Keyword(s):  
Protein Complex ◽  
Epitope Mapping ◽  
Super Resolution ◽  
Native Protein ◽  
Site Specific ◽  
Resolution Imaging

scholarly journals Diffusive Dynamics of NMDA Receptors in Live Neurons using Super-Resolution Imaging and Tracking

2018 ◽  
Vol 114 (3) ◽  
pp. 668a
Author(s):  
Chaoyi Jin ◽  
Sang Hak Lee ◽  
Phuong Le ◽  
Yeoan Youn ◽  
Pinghua Ge ◽  
...  
Keyword(s):  
Nmda Receptors ◽  
Super Resolution ◽  
Diffusive Dynamics ◽  
Resolution Imaging

Site-Specific Fluorescence Labeling of the β2Adrenergic Receptor Amino Terminus

1997 ◽  
Vol 254 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Anthony L. Parola ◽  
Sansan Lin ◽  
Brian K. Kobilka
Keyword(s):  
Fluorescence Labeling ◽  
Amino Terminus ◽  
Site Specific ◽  
Specific Fluorescence
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