Structural analysis of aliphatic versus aromatic substrate specificity in a copper amine oxidase from hansenula polymorpha

Valerie J. Klema, Corinne J. Solheid, Judith P. Klinman, Carrie M. Wilmot

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Copper amine oxidases (CAOs) are responsible for the oxidative deamination of primary amines to their corresponding aldehydes. The CAO catalytic mechanism can be divided into two half-reactions: a reductive half-reaction in which a primary amine substrate is oxidized to its corresponding aldehyde with the concomitant reduction of the organic cofactor 2,4,5-trihydroxyphenylalanine quinone (TPQ) and an oxidative half-reaction in which reduced TPQ is reoxidized with the reduction of molecular oxygen to hydrogen peroxide. The reductive half-reaction proceeds via Schiff base chemistry, in which the primary amine substrate first attacks the C5 carbonyl of TPQ, forming a series of covalent Schiff base intermediates. The X-ray crystal structures of copper amine oxidase-1 from the yeast Hansenula polymorpha (HPAO-1) in complex with ethylamine and benzylamine have been determined to resolutions of 2.18 and 2.25 Å, respectively. These structures reveal the two amine substrates bound at the back of the active site coincident with TPQ in its two-electron-reduced aminoquinol form. Rearrangements of particular amino acid side chains within the substrate channel and specific protein-substrate interactions provide insight into the substrate specificity of HPAO-1. These changes begin to account for this CAO's kinetic preference for small, aliphatic amines over the aromatic amines or whole peptides preferred by some of its homologues.

Original languageEnglish (US)
Pages (from-to)2291-2301
Number of pages11
JournalBiochemistry
Volume52
Issue number13
DOIs
StatePublished - Apr 2 2013
Externally publishedYes

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Amine Oxidase (Copper-Containing)
Pichia
Substrate Specificity
Structural analysis
Amines
Substrates
Schiff Bases
Aldehydes
Deamination
Molecular oxygen
Yeast
Hydrogen Peroxide
Catalytic Domain
Yeasts
Crystal structure
X-Rays
Electrons
Oxygen
Amino Acids
X rays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural analysis of aliphatic versus aromatic substrate specificity in a copper amine oxidase from hansenula polymorpha. / Klema, Valerie J.; Solheid, Corinne J.; Klinman, Judith P.; Wilmot, Carrie M.

In: Biochemistry, Vol. 52, No. 13, 02.04.2013, p. 2291-2301.

Research output: Contribution to journalArticle

Klema, Valerie J. ; Solheid, Corinne J. ; Klinman, Judith P. ; Wilmot, Carrie M. / Structural analysis of aliphatic versus aromatic substrate specificity in a copper amine oxidase from hansenula polymorpha. In: Biochemistry. 2013 ; Vol. 52, No. 13. pp. 2291-2301.
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