A tyrosine-derived free radical in apogalactose oxidase

Mei M. Whittaker, James Whittaker

Research output: Contribution to journalArticle

184 Citations (Scopus)

Abstract

Oxidation of apogalactose oxidase with ferricyanide leads to the formation of a stable free radical exhibiting distinctive optical absorption and EPR spectral features. The radical is associated with absorption in both near-UV and near-IR spectral regions, and its EPR spectrum is characteristic of an aromatic free radical with gav = 2.005. Reconstitution of both the apoenzyme and the free radical-containing form with copper substantially restores both the absorption spectra and the catalytic activity of the active enzyme, indicating that the preparation of the radical species does not significantly damage the protein. The absence of a free radical EPR signal in reconstituted and activated galactose oxidase containing nearly stoichiometric copper suggests the radical is an active site species relating to the free radical-coupled copper site previously proposed for this enzyme. Isotopic labeling experiments demonstrate that the radical derives from a tyrosine residue. The distinctive spectra associated with this radical indicate an environment which is different from that associated with the tyrosyl phenoxyl sites in other free radical enzymes.

Original languageEnglish (US)
Pages (from-to)9610-9613
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number17
StatePublished - Jun 15 1990
Externally publishedYes

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Free Radicals
Tyrosine
Oxidoreductases
Paramagnetic resonance
Copper
Enzymes
Galactose Oxidase
Apoenzymes
Light absorption
Labeling
Absorption spectra
Catalyst activity
Catalytic Domain
Oxidation
Proteins
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Whittaker, M. M., & Whittaker, J. (1990). A tyrosine-derived free radical in apogalactose oxidase. Journal of Biological Chemistry, 265(17), 9610-9613.

A tyrosine-derived free radical in apogalactose oxidase. / Whittaker, Mei M.; Whittaker, James.

In: Journal of Biological Chemistry, Vol. 265, No. 17, 15.06.1990, p. 9610-9613.

Research output: Contribution to journalArticle

Whittaker, MM & Whittaker, J 1990, 'A tyrosine-derived free radical in apogalactose oxidase', Journal of Biological Chemistry, vol. 265, no. 17, pp. 9610-9613.
Whittaker MM, Whittaker J. A tyrosine-derived free radical in apogalactose oxidase. Journal of Biological Chemistry. 1990 Jun 15;265(17):9610-9613.
Whittaker, Mei M. ; Whittaker, James. / A tyrosine-derived free radical in apogalactose oxidase. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 17. pp. 9610-9613.
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