Genetisch kodierte kupferfreie Klick‐Chemie

Genetically Encoded Cyclopropene Directs Rapid, Photoclick‐Chemistry‐Mediated Protein Labeling in Mammalian Cells

“…[5] Numerous small organic groups such as ketone, [6] azides, [7] terminal alkynes, [8] and terminal alkenes, [9] as well as larger reactive bioorthogonal groups such as cyclooctyne, [10] norbornene, [11] transcyclooctene, [10b, 11b] tetrazole, [12] and tetrazine [13] have been genetically encoded for site-selective protein labeling in vivo. To track fast protein dynamics in vivo, it is imperative that these genetically encoded bioorthogonal reporters direct fast and selective bioorthogonal labeling with the cognate biophysical probes, preferably with a spatiotemporal control.…”

“…Compared to other genetically encoded, bioorthogonal labeling reactions reported recently, [10–13] the main advantage of the cyclopropene-directed photoclick chemistry lies in its potential in the spatiotemporally controlled protein labeling in mammalian cells, which requires the development of highly reactive laser-activatable tetrazole reagents using either single photon (e.g. 405 nm) or two-photon laser source; work along this line is currently in progress.…”

Protein Spin Labeling with a Photocaged Nitroxide Using Diels-Alder Chemistry

“…TRX was the first protein to be modified by aD A inv reaction in 2008, [16a] and previousE PR studies are available for data comparison. [10e, 30] Both model proteins exhibit native cysteiner esidues;t husp roviding the possibility to test for orthogonal labeling without affecting cysteine.T he first ncAA that could undergo aD A inv reaction, al ysine-derived cyclooctyne, was introduced for the copper-free click reaction in 2011, [31] before at etrazine, [32] norbornenes, [15a, 33] TCOs, [15a] and aspirohexene [34] were also genetically encoded. The rationally designed Methanosarcina mazei mutant tRNA Pyl /PylRS AF (Y306A, Y384F) possesses an enlarged binding pocket that is suitable fori ncorporating these bulky ncAAs in responset ot he amber stop codon.…”

The Power of Fluorogenic Probes

“…Thus, several techniques have been developed very recently in which a specific artificial amino acid is treated with a fluorogenic dye. [5] When it comes to fluorescence labeling inside cells, it is generally desirable to have the bioorthogonally reactive group directly involved in the fluorophore quenching (Figure 1…”