Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase

Mei M. Whittaker, Philip J. Kersten, Nobuhumi Nakamura, Joann Sanders-Loehr, Elizabeth S. Schweizer, James Whittaker

Research output: Contribution to journalArticle

164 Citations (Scopus)

Abstract

A free radical-coupled copper complex has been identified as the catalytic structure in the active site of glyoxal oxidase from Phanerochaete chrysosporium based on a combination of spectroscopic and biochemical studies. The native (inactive) enzyme is activated by oxidants leading to the elimination of the cupric EPR signal consistent with formation of an antiferromagnetically coupled radical-copper complex. Oxidation also leads to the appearance of a substoichiometric free radical EPR signal with an average g value (gav = 2.0055) characteristic of phenoxyl π-radicals arising from a minority apoenzyme fraction. Optical absorption, CD, and spectroelectrochemical measurements on the active enzyme reveal complex spectra extending into the near IR and define the redox potential for radical formation (E1/2 = 0.64 V versus NHE, pH 7.0). Resonance Raman spectra have identified the signature of a modified (cysteinyl-tyrosine) phenoxyl in the vibrational spectra of the active complex. This radical-copper motif has previously been found only in galactose oxidase, with which glyoxal oxidase shares many properties despite lacking obvious sequence identity, and catalyzing a distinct reaction. The enzymes thus represent members of a growing class of free radical metalloenzymes based on the radical-copper catalytic motif and appear to represent functional variants that have evolved to distinct catalytic roles.

Original languageEnglish (US)
Pages (from-to)681-687
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number2
StatePublished - Jan 12 1996
Externally publishedYes

Fingerprint

Phanerochaete
Copper
Free Radicals
Paramagnetic resonance
Enzymes
Galactose Oxidase
Apoenzymes
Vibrational spectra
Oxidants
Light absorption
Oxidation-Reduction
Tyrosine
Raman scattering
Catalytic Domain
Oxidation
copper oxidase
glyoxal oxidase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Whittaker, M. M., Kersten, P. J., Nakamura, N., Sanders-Loehr, J., Schweizer, E. S., & Whittaker, J. (1996). Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase. Journal of Biological Chemistry, 271(2), 681-687.

Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase. / Whittaker, Mei M.; Kersten, Philip J.; Nakamura, Nobuhumi; Sanders-Loehr, Joann; Schweizer, Elizabeth S.; Whittaker, James.

In: Journal of Biological Chemistry, Vol. 271, No. 2, 12.01.1996, p. 681-687.

Research output: Contribution to journalArticle

Whittaker, MM, Kersten, PJ, Nakamura, N, Sanders-Loehr, J, Schweizer, ES & Whittaker, J 1996, 'Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase', Journal of Biological Chemistry, vol. 271, no. 2, pp. 681-687.
Whittaker MM, Kersten PJ, Nakamura N, Sanders-Loehr J, Schweizer ES, Whittaker J. Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase. Journal of Biological Chemistry. 1996 Jan 12;271(2):681-687.
Whittaker, Mei M. ; Kersten, Philip J. ; Nakamura, Nobuhumi ; Sanders-Loehr, Joann ; Schweizer, Elizabeth S. ; Whittaker, James. / Glyoxal oxidase from Phanerochaete chrysosporium is a new radical-copper oxidase. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 2. pp. 681-687.
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