2015
DOI: 10.1021/acs.bioconjchem.5b00424
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Synthesis and Incorporation of Unnatural Amino Acids To Probe and Optimize Protein Bioconjugations

Abstract: The utilization of unnatural amino acids (UAAs) in bioconjugations is ideal due to their ability to confer a degree of bioorthogonality and specificity. In order to elucidate optimal conditions for the preparation of bioconjugates with UAAs, we synthesized 9 UAAs with variable methylene tethers (2–4) and either an azide, alkyne, or halide functional group. All 9 UAAs were then incorporated into green fluorescent protein (GFP) using a promiscuous aminoacyl-tRNA synthetase. The different bioconjugations were the… Show more

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Cited by 19 publications
(24 citation statements)
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“…The most common bioorthogonal ncAAs that have been encoded have keto, 200 azido 59 and acetylenic side chains (Figure 8). 42, 63, 201, 202 Azide and alkynyl ncAAs undergo highly selective copper-catalyzed 1,3-dipolar cycloaddition reactions and have been generally used for in vitro applications. 203, 204 However, strained alkynes have recently been genetically encoded to afford copper-free “click” reactions that can be used in living cells.…”
Section: Applications Of Non-canonical Amino Acidsmentioning
confidence: 99%
“…The most common bioorthogonal ncAAs that have been encoded have keto, 200 azido 59 and acetylenic side chains (Figure 8). 42, 63, 201, 202 Azide and alkynyl ncAAs undergo highly selective copper-catalyzed 1,3-dipolar cycloaddition reactions and have been generally used for in vitro applications. 203, 204 However, strained alkynes have recently been genetically encoded to afford copper-free “click” reactions that can be used in living cells.…”
Section: Applications Of Non-canonical Amino Acidsmentioning
confidence: 99%
“…For example (Scheme 2), alkylation with bromide 29 gave access to compound 7 by deprotection, compound 8 by Sonogashira coupling, compound 9 by phthalimide deprotection, Gln residue coupling, and Boc deprotection, and compound 10 by Sonagashira coupling using alkyne 31 , prepared from Boc-Tyr methyl ester in two steps. 27 For the full structures of compounds 7, 9 , and 10 , please see Figure 4.…”
Section: Resultsmentioning
confidence: 99%
“…NBS, CCl 4 , 62%; b. NaH, KI, DMF, 80%; c. TBAF, THF, 88%; d. N-phthaloyl 1-amino-4-pentyne, 29 Pd(PPh 3 ) 4 , pyrrolidine, 42%; e. H 2 NNH 2 -H 2 O, EtOH,; then N-Boc-Gln, HATU, DIPEA, DMF, 21%; then 4M HCl in dioxane, 97%; f) Pd(PPh 3 ) 4 , pyrrolidine, 80°C, 42%; then 4N HCl in dioxane, 51%; g) 5-chloro-1-pentyne, K 2 CO 3 , DMF, 97%; then LiOH-H 2 O, MeOH-THF, 99%.…”
Section: Figurementioning
confidence: 99%
“…1821 More recently, our group has developed a bioorthogonal variant of the Glaser-Hay reaction, 22,23 which brings together two terminal alkynes to form a diyne in the presence of copper(I) under physiological conditions. 2426 The resulting stable diyne linkage has a well-defined linear geometry, and due to the abundance of commercially available terminal alkynes, a variety of chemical moieties can be reacted onto a protein using this technique. As such, we sought to utilize the power of this new chemistry to generate new and different protein function dependent upon the alkyne reaction partner and not the UAA alone.…”
mentioning
confidence: 99%