PMID: 108101Feb 15, 1979

The enzyme-bound copper of dopamine beta-monooxygenase. Reaction with copper chelators, preparation of the apoprotein, and kinetics of the reconstitution by added copper

European Journal of Biochemistry
T Skotland, T Ljones

Abstract

The enzyme-bound copper of dopamine beta-monooxygenase reacted rapidly with the chelator bathocuproine disulfonate; the reaction in the presence of ascorbate was completed in 2 min at 25 degrees C with 1mM chelator. This reaction and also the reaction with EDTA could be used to prepare the apoenzyme, which in both cases was completely reactivated in less than 10 s. The reactivation data gave apparent Michaelis constants for copper 0.03 -- 0.2 micron. Trace amounts of copper in buffers and assay mixtures gave significant reactivation without added copper, unless they had been treated with a chelating resin. Titrations using the different chelation rates of free and enzyme-bound copper indicated that four copper atoms are bound per enzyme molecule of four subunits. The native enzyme was more stable against thermal inactivation than the apoenzyme, but this stability was only partially restored by addition of copper to the apoenzyme.

References

Jan 1, 1976·Advances in Experimental Medicine and Biology·H S Mason
Jan 16, 1978·Clinica Chimica Acta; International Journal of Clinical Chemistry·Y EshelS Gitter
Feb 21, 1977·Biochemical and Biophysical Research Communications·T SkotlandK Sletten
Jan 15, 1976·European Journal of Biochemistry·T LjonesT Flatmark
Aug 1, 1967·Analytical Biochemistry·M Van de Bogart, H Beinert
Jan 1, 1969·European Journal of Biochemistry·R MalkinT Vänngård
Dec 4, 1972·Biochemical and Biophysical Research Communications·D KerteszE Antonini
Dec 1, 1974·FEBS Letters·T Ljones, T Flatmark
Aug 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·E F WallaceW Lovenberg
Jun 1, 1974·The Biochemical Journal·R Eisenthal, A Cornish-Bowden
Feb 1, 1966·Biochemistry·N E GoodR M Singh

Citations

Jul 15, 1984·Experientia·J L Dreyer
Mar 22, 2001·Journal of Neurochemistry·P MegawM Boelen
Jan 1, 1982·Scandinavian Journal of Immunology. Supplement·H Winkler

Related Concepts

Adrenal Medulla
Apoenzymes
Bos indicus
Metal Antagonists
Copper
Dopamine-beta-monooxygenase
Edetic Acid, Sodium Salt
Oxydol
Spectrometry
Structure-Activity Relationship

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