Development of Strategies for the Site-Specific In Vivo Incorporation of Photoreactive Amino Acids: p-Azidophenylalanine, p-Acetylphenylalanine and Benzofuranylalanine

Behrens, Carsten; Nielsen, Joachim; Fan, Xuejun; Doisy, Xavier; Kim, Ki‐Hyun; Prætorius-Ibba, Mette; Nielsen, Peter E.; Ibba, Michael

doi:10.1016/s0040-4020(00)00828-0

Cited by 18 publications

(14 citation statements)

References 27 publications

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“…Synthesis of Bicyclic Amino Acids-Benzofuranylalanine was synthesized as previously reported (13), and the novel benzotriazolylalanine was prepared as shown in Scheme 1 starting from commercially available 5-(hydroxymethyl)triazole using standard chemistry. Enantiomeric resolution was achieved using Carlsberg subtilisin (13) for specific hydrolysis of the L-form of an ester intermediate.…”

Section: Resultsmentioning

confidence: 99%

“…The organic phase was evaporated in vacuo to an oil, which was dissolved in water (100 ml) and freeze-dried.5XZ was obtained as a white powder (840 mg, 40%). FAB-MS m/z 339 (MϩH) data: 1 H NMR (CD 3 OD): ␦ 7.9 -7.2 (m, 16H), 5.9 (s, 4H), 4.7 (m, 2H), 3.4-3.0 (m, 4H), 1.9 and 1.7 (m, 6H); 13 …”

Section: N-acetyl-3-(n-benzyl-benzotriazole-5-yl)-dl-alanine Ethyl Esmentioning

confidence: 99%

“…The residue was triturated with diethyl ether, and the product was obtained as a white powder (400 mg, 96%). FAB-MS m/z 207 (MϩH) data: 1 H NMR (CD 3 OD): ␦ 8.05 (m, 2H), 7.74 (d, 1H, J ϭ 9.1 Hz), 4.47 (t, 1H, J ϭ 6.5 Hz), 3.7-3.4 (ABX-system, J AB ϭ 17.5 Hz, J AX ϭ 6.0 Hz, J BX ϭ 7.2 Hz); 13 UV Irradiation-Aqueous solutions of benzofuranylalanine and benzotriazolylalanine at the indicated concentrations in a quartz cuvette sealed with Parafilm were placed immediately adjacent to the desired UV light source with the following characteristics: Struers UVG-11, 254 nm; Philips TL20W/12RS, 320 nm; Phillips 20 W/09N, 365 nm. Samples were irradiated at room temperature for the desired length of time.…”

Section: L-3-(benzotriazole-5-yl)-l-alanine Hydrochloride (7)-compoundmentioning

confidence: 99%

“…Based upon previous studies of proteinogenic photoreactive amino acid analogues with bicyclic structures containing a benzene ring "scaffold" (13), we set out to test such compounds as substrates for incorporation into a protein via PheRS-␣A294G. Other unnatural amino acids shown to be substrates for this aminoacyl tRNA synthetase variant display distinct yet limited structural divergence from phenylalanine.…”

mentioning

confidence: 99%

“…Although PheRS-␣A294G (and PheRS-␣T251G/A294G) appear rather flexible with respect to modifications at the para position of the benzene side chain, it is far from obvious that this mutant would accommodate amino acids carrying double ring systems involving joint substitutions at the meta and para position of the phenylalanine benzene side chain. Nevertheless, based on growth inhibition, we have previously proposed that benzofuranylalanine is indeed a substrate for PheRS-␣A294G, opening up the possibility that such bicyclic amino acid analogs could be developed as substrates for protein synthesis (13).…”

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confidence: 99%

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Photoreactive Bicyclic Amino Acids as Substrates for Mutant Escherichia coli Phenylalanyl-tRNA Synthetases

Bentin

Hamzavi

Salomonsson

et al. 2004

Journal of Biological Chemistry

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Unnatural amino acids carrying reactive groups that can be selectively activated under non-invasive biologically benign conditions are of interest in protein engineering as biological tools for the analysis of proteinprotein and protein-nucleic acids interactions. The double ring system phenylalanine analogues benzofuranylalanine and benzotriazolylalanine were synthesized, and their photolability was tested by UV irradiation at 254, 320, and 365 nm. Although both showed photo reactivity, benzofuranylalanine appeared as the most promising compound because this amino acid was activated by UVA (long wavelength) irradiation. These amino acids were also tested for in vitro charging of tRNA Phe and for protein mutagenesis via the phenylalanyl-tRNA synthetase variant ␣A294G that is able to facilitate in vivo protein synthesis using a range of para-substituted phenylalanine analogues. The results demonstrate that benzofuranylalanine, but not benzotriazolylalanine, is a substrate for phenylalanine tRNA synthetase ␣A294G, and matrix-assisted laser desorption ionization time-offlight analysis showed it to be incorporated into a model protein with high efficiency. The in vivo incorporation into a target protein of a bicyclic phenylalanine analogue, as described here, demonstrates the applicability of phenylalanine tRNA synthetase variants in expanding the scope of protein engineering.

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Section: Resultsmentioning

confidence: 99%

Section: N-acetyl-3-(n-benzyl-benzotriazole-5-yl)-dl-alanine Ethyl Esmentioning

confidence: 99%

Section: L-3-(benzotriazole-5-yl)-l-alanine Hydrochloride (7)-compoundmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Photoreactive Bicyclic Amino Acids as Substrates for Mutant Escherichia coli Phenylalanyl-tRNA Synthetases

Bentin

Hamzavi

Salomonsson

et al. 2004

Journal of Biological Chemistry

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The Krapcho Dealkoxycarbonylation Reaction of Esters with α‐Electron‐Withdrawing Substituents

Krapcho

Ciganek

2013

Organic Reactions

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The Krapcho reaction involves esters with α‐electron‐withdrawing substituents such as malonates, β‐keto esters, and α‐cyano esters, which undergo dealkoxycarbonylation on being heated in polar aprotic solvents (such as DMSO, DMF, or HMPT) in the presence of water, or in polar aprotic solvents with water in the presence of added salts (such as NaCN, NaCl, LiCl, LiI, or MgCl 2 ). This procedure avoids the use of strongly aqueous acidic and alkaline conditions and tolerates many functional and protecting groups. The chapter presents the mechanistic aspects of this procedure, which include reaction parameters such as solvents, salts, other additives, and the use of microwave irradiation. The diastereoselectivity of protonation of the enolate intermediate leading to the dealkoxycarbonylated products is discussed. Trapping of the intermediate enolate by electrophiles other than a proton is illustrated. Potential side reactions and same‐pot subsequent reactions of substrates are discussed along with functional‐group compatibility for halogens and nitrogen, oxygen, sulfur, selenium, and carbon functional groups. Several examples of applications of this methodology for the synthesis of natural products along with experimental conditions and procedures are presented. Closely related methods are discussed in the Comparison with Other Methods section. The tables, which consist of 371 pages, are grouped according to substrate structure. Selective entries for other methods (excluding classical aqueous acidic or basic methods) are included in the tabular survey for comparative purposes.

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ChemInform Abstract: Development of Strategies for the Site‐Specific in vivo Incorporation of Photoreactive Amino Acids: p‐Azidophenylalanine, p‐Acetylphenylalanine and Benzofuranylalanine.

Behrens¹,

Nielsen²,

Fan³

et al. 2001

ChemInform

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Development of Strategies for the Site-Specific In Vivo Incorporation of Photoreactive Amino Acids: p-Azidophenylalanine, p-Acetylphenylalanine and Benzofuranylalanine

Cited by 18 publications

References 27 publications

Photoreactive Bicyclic Amino Acids as Substrates for Mutant Escherichia coli Phenylalanyl-tRNA Synthetases

Photoreactive Bicyclic Amino Acids as Substrates for Mutant Escherichia coli Phenylalanyl-tRNA Synthetases

The Krapcho Dealkoxycarbonylation Reaction of Esters with α‐Electron‐Withdrawing Substituents

ChemInform Abstract: Development of Strategies for the Site‐Specific in vivo Incorporation of Photoreactive Amino Acids: p‐Azidophenylalanine, p‐Acetylphenylalanine and Benzofuranylalanine.

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