Synthesis and Evaluation of Photoreactive Tetrazole Amino Acids

Wang, Yizhong; Lin, Qing

doi:10.1021/ol901300h

“…To this end, a two-step procedure is employed: a bioorthogonal tag is first introduced into the protein of interest site-specifically using amber codon suppression technique [22]; and then a biophysical probe carrying the cognate functionality is allowed to react selectively with the pre-tagged protein via the corresponding bioorthogonal reaction. For the application of photoclick chemistry to protein labeling, one can either genetically encode a tetrazole moiety [23] or an alkene dipolarophile. Although p -(2-tetrazole)phenylalanine has been incorporated site-selectively into the proteins [24] using an engineered M. jannaschii tyrosyl-tRNA synthetase/ Mj tRNA CUA pair, the photoclick reaction with a fumarate-modified fluorescein was slow ( k 2 = 0.082 ± 0.011 M −1 s −1 in 1:1 ACN/PBS) because of the reduced reactivity of the monoaryltetrazole.…”

Section: Site-specific Protein Labelingmentioning

confidence: 99%

Photoclick chemistry: a fluorogenic light-triggered in vivo ligation reaction

Ramil

¹

,

Lin

²

2014

Current Opinion in Chemical Biology

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The ability to use chemical reactivity to monitor and control biomolecular processes with a spatial and temporal precision motivated the development of light-triggered in vivo chemistries. To this end, the photoinduced tetrazole-alkene cycloaddition, also termed “photoclick chemistry” offers a very rapid chemical ligation platform for the manipulation of biomolecules and matrices in vivo. Here we outline the recent developments in the optimization of this chemistry, ranging from the search for substrates that offer two-photon photoactivatability, superior reaction kinetics, and/or genetic encodability, to the study of the reaction mechanism. The applications of the photoclick chemistry in protein labeling in vitro and in vivo as well as in preparing “smart” hydrogels for 3D cell culture are highlighted.

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“…One approach was to incorporate the tetrazole moiety into the amino acid side chain and evolve orthogonal aminoacyl-tRNA synthetase/tRNA pairs to charge the resulting tetrazole amino acids into proteins site-specifically. To this end, we synthesized a series of tetrazole modified unnatural amino acids (Scheme 8b), and tested their reactivity in the photo-triggered cycloaddition reaction with methyl methacrylate in PBS/MeCN (1:1) [43]. While four out of six tetrazole amino acids gave excellent reaction yields, only p -tetrazole-phenylalanine ( p -Tpa ) was incorporated into myoglobin site-specifically using an evolved synthetase via the amber codon suppression [44].…”

Section: Photoinduced 13-dipolar Cycloaddition Reaction Between Tementioning

confidence: 99%

Photo-Triggered Click Chemistry for Biological Applications

Herner

¹

,

Lin

²

2015

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In the last decade and a half, numerous bioorthogonal reactions have been developed with a goal to study biological processes in their native environment, i.e., in living cells and animals. Among them, the photo-triggered reactions offer several unique advantages including operational simplicity with the use of light rather than toxic metal catalysts and ligands, and exceptional spatiotemporal control through the application of an appropriate light source with pre-selected wavelength, light intensity and exposure time. While the photoinduced reactions have been studied extensively in materials research, e.g., on macromolecular surface, the adaptation of these reactions for chemical biology applications is still in its infancy. In this chapter, we review the recent efforts in the discovery and optimization the photo-triggered bioorthogonal reactions, with a focus on those that have shown broad utility in biological systems. We discuss in each cases the chemical and mechanistic background, the kinetics of the reactions and the biological applicability together with the limiting factors.

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“…One approach was to incorporate the tetrazole moiety into the amino acid side chain and evolve orthogonal aminoacyl-tRNA synthetase/tRNA pairs to charge the resulting tetrazole amino acids into proteins site-specifically. To this end, we synthesized a series of tetrazole modified unnatural amino acids (Scheme 8b), and tested their reactivity in the photo-triggered cycloaddition reaction with methyl methacrylate in PBS/MeCN (1:1) [43]. While four out of six tetrazole amino acids gave excellent reaction yields, only p-tetrazole-phenylalanine (p-Tpa) was incorporated into myoglobin sitespecifically using an evolved synthetase via the amber codon suppression [44].…”

Section: Site-specific Labeling Of Proteins Via the Photoinduced Tetrmentioning

confidence: 99%

Photo-Triggered Click Chemistry for Biological Applications

Herner¹,

Qi²

2015

Cycloadditions in Bioorthogonal Chemistry

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In the last decade and a half, numerous bioorthogonal reactions have been developed with a goal to study biological processes in their native environment, i.e., in living cells and animals. Among them, the photo-triggered reactions offer several unique advantages including operational simplicity with the use of light rather than toxic metal catalysts and ligands, and exceptional spatiotemporal control through the application of an appropriate light source with pre-selected wavelength, light intensity and exposure time. While the photoinduced reactions have been studied extensively in materials research, e.g., on macromolecular surface, the adaptation of these reactions for chemical biology applications is still in its infancy. In this chapter, we review the recent efforts in the discovery and optimization the photo-triggered bioorthogonal reactions, with a focus on those that have shown broad utility in biological systems. We discuss in each cases the chemical and mechanistic background, the kinetics of the reactions and the biological applicability together with the limiting factors.

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Synthesis and Evaluation of Photoreactive Tetrazole Amino Acids

Cited by 50 publications

References 19 publications

Photoclick chemistry: a fluorogenic light-triggered in vivo ligation reaction

Photoclick chemistry: a fluorogenic light-triggered in vivo ligation reaction

Photo-Triggered Click Chemistry for Biological Applications

Photo-Triggered Click Chemistry for Biological Applications

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