Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR

G. Reid Bishop, Zhenyu Zhu, Tracy L. Whitehead, Rickey Paige Hicks, Victor L. Davidson

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

13C- and 15N-NMR studies of the reaction of aromatic amine dehydrogenase (AADH) with methylamine demonstrated that the products of the reductive half-reaction are an equivalent of formaldehyde hydrate and a reduced aminoquinol form of the tryptophan tryptophylquinone (TTQ) cofactor which contains covalently bound substrate-derived N. These data are consistent with the Ping Pong kinetic mechanism and aminotransferase-type chemical reaction mechanism which have been previously proposed for AADH. Comparison of the 15N-NMR spectra of the aminoquinol TTQ intermediates of AADH and methylamine dehydrogenase (MADH) revealed that the substrate-derived aminoquinol N of AADH and MADH exhibited distinct 15N chemical shifts which are separated by approx. 7 p.p.m. In each case, the signal for the substrate-derived aminoquinol N appears optimally with short pulse delay and exhibits a relaxation time and chemical shift which are consistent with 15N covalently bound to an aromatic ring (i.e. aminoquinol) which is attached to a rigid protein matrix. The aminoquinol of AADH is less stable against reoxidation than that of MADH. These data suggest that differences in the active-site mediated electrostatic environments of the aminoquinol N in the respective enzymes may influence both the observed 15N chemical shift and the relative reactivities of the TTQ aminoquinols towards oxygen. These data also demonstrate the utility of 13C- and 15N-NMR spectroscopy as a tool for monitoring the intermediates and products of enzyme-catalysed transformations.

Original languageEnglish (US)
Pages (from-to)1159-1163
Number of pages5
JournalBiochemical Journal
Volume330
Issue number3
DOIs
StatePublished - Mar 15 1998

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Reaction intermediates
Reaction products
Nuclear magnetic resonance
Chemical shift
Substrates
Enzymes
Transaminases
Hydrates
Static Electricity
Relaxation time
Formaldehyde
Nuclear magnetic resonance spectroscopy
Chemical reactions
Electrostatics
Catalytic Domain
Magnetic Resonance Spectroscopy
aromatic amine dehydrogenase
Carbon-13 Magnetic Resonance Spectroscopy
Oxygen
Kinetics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Bishop, G. R., Zhu, Z., Whitehead, T. L., Hicks, R. P., & Davidson, V. L. (1998). Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR. Biochemical Journal, 330(3), 1159-1163. https://doi.org/10.1042/bj3301159

Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR. / Bishop, G. Reid; Zhu, Zhenyu; Whitehead, Tracy L.; Hicks, Rickey Paige; Davidson, Victor L.

In: Biochemical Journal, Vol. 330, No. 3, 15.03.1998, p. 1159-1163.

Research output: Contribution to journalArticle

Bishop, GR, Zhu, Z, Whitehead, TL, Hicks, RP & Davidson, VL 1998, 'Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR', Biochemical Journal, vol. 330, no. 3, pp. 1159-1163. https://doi.org/10.1042/bj3301159
Bishop, G. Reid ; Zhu, Zhenyu ; Whitehead, Tracy L. ; Hicks, Rickey Paige ; Davidson, Victor L. / Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR. In: Biochemical Journal. 1998 ; Vol. 330, No. 3. pp. 1159-1163.
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