Examination of the new α-(2′Z-fluoro)vinyl trigger with lysine decarboxylase

The absolute stereochemistry dictates the reaction course

Kannan R. Karukurichi, Roberta De La Salud-Bea, Wan Jin Jahng, David B Berkowitz

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The first examination of a terminal α-fluorovinyl trigger in an amino acid decarboxylase active site is reported. To investigate the enantiospecificity of inactivation with this new AADC trigger, an enantioselective synthesis of l-α-(2′Z-fluoro)vinyllysine and its d-antipode has been developed. Control of stereochemistry is achieved through introduction of the amino acid side chain via alkylation of a chiral vinylglycine-derived dienolate. Facial selectivity is conferred by a trans-2′(β-naphthyl)-2′-propylcyclohexyl ester auxiliary, available in both antipodal forms (Comins protocol). The alkylation employs a new electrophile, N-p-methoxybenzyl-N-(2′-trimethylsilylethanesulfonyl)-4-iodobutylamine, for convergent installation of the lysine side chain. Vinyl to 2′-fluorovinyl interconversion then provides l- and d-α-(2′Z-fluoro)vinyllysine in 97-99% ee, as demonstrated by chiral HPLC. Both time-dependent enzyme kinetics and 19F NMR reveal striking differences in the behavior of these two antipodes in the lysine decarboxylase active site. The l-antipode displays time dependent inactivation (t1/2 = 3 ± 1 min; KI = 86 ± 22 μM), whereas the d-antipode behaves as a substrate, being completely turned over to α-(2′Z-fluoro)vinylcadaverine. Titration of LDC with varying amounts of l-α-(2′Z-fluoro)vinyllysine provides an estimate of 20 ± 3 for the partition ratio for this antipode. 19F NMR provides a more detailed account of the inactivation with the l-antipode, revealing that 1 in 3.4 turnovers of this mechanism-based inhibitor results in errant protonation (required by design), with 1 in 5 errant protonation events leading to LDC inactivation. This gives an overall partition ratio of 16 ± 2. Fluoride-selective electrode measurements are in agreement with 19F NMR estimates of [fluoride] released.

Original languageEnglish (US)
Pages (from-to)258-259
Number of pages2
JournalJournal of the American Chemical Society
Volume129
Issue number2
DOIs
StatePublished - Jan 17 2007

Fingerprint

lysine decarboxylase
Stereochemistry
Alkylation
Fluorides
Catalytic Domain
Protonation
Nuclear magnetic resonance
Amino Acids
Carboxy-Lyases
Amino acids
Lysine
Enzyme kinetics
Electrodes
Esters
High Pressure Liquid Chromatography
Titration
Enzymes
Substrates

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Examination of the new α-(2′Z-fluoro)vinyl trigger with lysine decarboxylase : The absolute stereochemistry dictates the reaction course. / Karukurichi, Kannan R.; De La Salud-Bea, Roberta; Jahng, Wan Jin; Berkowitz, David B.

In: Journal of the American Chemical Society, Vol. 129, No. 2, 17.01.2007, p. 258-259.

Research output: Contribution to journalArticle

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abstract = "The first examination of a terminal α-fluorovinyl trigger in an amino acid decarboxylase active site is reported. To investigate the enantiospecificity of inactivation with this new AADC trigger, an enantioselective synthesis of l-α-(2′Z-fluoro)vinyllysine and its d-antipode has been developed. Control of stereochemistry is achieved through introduction of the amino acid side chain via alkylation of a chiral vinylglycine-derived dienolate. Facial selectivity is conferred by a trans-2′(β-naphthyl)-2′-propylcyclohexyl ester auxiliary, available in both antipodal forms (Comins protocol). The alkylation employs a new electrophile, N-p-methoxybenzyl-N-(2′-trimethylsilylethanesulfonyl)-4-iodobutylamine, for convergent installation of the lysine side chain. Vinyl to 2′-fluorovinyl interconversion then provides l- and d-α-(2′Z-fluoro)vinyllysine in 97-99{\%} ee, as demonstrated by chiral HPLC. Both time-dependent enzyme kinetics and 19F NMR reveal striking differences in the behavior of these two antipodes in the lysine decarboxylase active site. The l-antipode displays time dependent inactivation (t1/2 = 3 ± 1 min; KI = 86 ± 22 μM), whereas the d-antipode behaves as a substrate, being completely turned over to α-(2′Z-fluoro)vinylcadaverine. Titration of LDC with varying amounts of l-α-(2′Z-fluoro)vinyllysine provides an estimate of 20 ± 3 for the partition ratio for this antipode. 19F NMR provides a more detailed account of the inactivation with the l-antipode, revealing that 1 in 3.4 turnovers of this mechanism-based inhibitor results in errant protonation (required by design), with 1 in 5 errant protonation events leading to LDC inactivation. This gives an overall partition ratio of 16 ± 2. Fluoride-selective electrode measurements are in agreement with 19F NMR estimates of [fluoride] released.",
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N2 - The first examination of a terminal α-fluorovinyl trigger in an amino acid decarboxylase active site is reported. To investigate the enantiospecificity of inactivation with this new AADC trigger, an enantioselective synthesis of l-α-(2′Z-fluoro)vinyllysine and its d-antipode has been developed. Control of stereochemistry is achieved through introduction of the amino acid side chain via alkylation of a chiral vinylglycine-derived dienolate. Facial selectivity is conferred by a trans-2′(β-naphthyl)-2′-propylcyclohexyl ester auxiliary, available in both antipodal forms (Comins protocol). The alkylation employs a new electrophile, N-p-methoxybenzyl-N-(2′-trimethylsilylethanesulfonyl)-4-iodobutylamine, for convergent installation of the lysine side chain. Vinyl to 2′-fluorovinyl interconversion then provides l- and d-α-(2′Z-fluoro)vinyllysine in 97-99% ee, as demonstrated by chiral HPLC. Both time-dependent enzyme kinetics and 19F NMR reveal striking differences in the behavior of these two antipodes in the lysine decarboxylase active site. The l-antipode displays time dependent inactivation (t1/2 = 3 ± 1 min; KI = 86 ± 22 μM), whereas the d-antipode behaves as a substrate, being completely turned over to α-(2′Z-fluoro)vinylcadaverine. Titration of LDC with varying amounts of l-α-(2′Z-fluoro)vinyllysine provides an estimate of 20 ± 3 for the partition ratio for this antipode. 19F NMR provides a more detailed account of the inactivation with the l-antipode, revealing that 1 in 3.4 turnovers of this mechanism-based inhibitor results in errant protonation (required by design), with 1 in 5 errant protonation events leading to LDC inactivation. This gives an overall partition ratio of 16 ± 2. Fluoride-selective electrode measurements are in agreement with 19F NMR estimates of [fluoride] released.

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