Full, reversible copper removal from ascorbate oxidase

Anaerobic treatment with cyanide of reduced ascorbate oxidase causes total depletion of copper. No significant amount of the metal is reincorporated when the apo-enzyme is incubated with cupric ions, but it is upon incubation with a stoichiometric amount (eight mol per mol of native enzyme) of a Cu(...

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Bibliographic Details
Published in:	Biochemical and biophysical research communications Vol. 131; no. 3; p. 1251
Main Authors:	Savini, I, Morpurgo, L, Avigliano, L
Format:	Journal Article
Language:	English
Published:	United States 30-09-1985
Subjects:	Apoenzymes Apoproteins Ascorbate Oxidase Chemical Phenomena Chemistry Copper - isolation & purification Cyanides Electron Spin Resonance Spectroscopy Organometallic Compounds Oxidoreductases Oxygen Spectrophotometry Thiourea - analogs & derivatives
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Summary:	Anaerobic treatment with cyanide of reduced ascorbate oxidase causes total depletion of copper. No significant amount of the metal is reincorporated when the apo-enzyme is incubated with cupric ions, but it is upon incubation with a stoichiometric amount (eight mol per mol of native enzyme) of a Cu(I) complex stable in air [Cu(I)(thiourea)3]Cl. The yield in reconstituted protein is higher under anaerobic conditions (85-90%) than in air (70-75%). By treatment with less than stoichiometric amounts of [Cu(I)(thiourea)3]Cl the apo-protein binds copper preferentially at the blue copper site. As a consequence the recovery of enzymatic activity is percentually lower than copper reincorporation.
ISSN:	0006-291X
DOI:	10.1016/0006-291X(85)90225-6