Mechanism of the tyrosine ammonia lyase reaction-tandem nucleophilic and electrophilic enhancement by a proton transfer

Sarolta Pilbák, Ödön Farkas, László Poppe

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25 Citations (Scopus)

Abstract

Quantum mechanics/molecular mechanics calculations in tyrosine ammonia lyase (TAL) ruled out the hypothetical Friedel-Crafts (FC) route for ammonia elimination from l-tyrosine due to the high energy of FC intermediates. The calculated pathway from the zwitterionic l-tyrosine-binding state (0.0 kcalmol -1) to the productbinding state ((E)-coumarate+H 2N-MIO;-24.0 kcalmol -1; MIO=3,5-dihydro-5-methylidene-4H- imidazol-4-one) involves an intermediate (IS,-19.9 kcalmol -1), which has a covalent bond between the N atom of the substrate and MIO, as well as two transition states (TS1 and TS2). TS1 (14.4 kcalmol-1) corresponds to a proton transfer from the substrate to the N1 atom of MIO by Tyr300-OH. Thus, a tandem nucleophilic activation of the substrate and electrophilic activation of MIO happens. TS2 (5.2 kcal mol-1) indicates a concerted C-N bond breaking of the N-MIO intermediate and deprotonation of the pro-S b position by Tyr60. Calculations elucidate the role of enzymic bases (Tyr60 and Tyr300) and other catalytically relevant residues (Asn203, Arg303, and Asn333, Asn435), which are fully conserved in the amino acid sequences and in 3D structures of all known MIO-containing ammonia lyases and 2,3-aminomutases.

Original languageEnglish
Pages (from-to)7793-7802
Number of pages10
JournalChemistry - A European Journal
Volume18
Issue number25
DOIs
Publication statusPublished - Jun 18 2012

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Keywords

  • Conformation analysis
  • Density functional calculations
  • Enzyme catalysis
  • Lyases
  • Reaction mechanisms

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

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