Abstract
The reaction of yeast Cu-MT with nitric oxide (NO) was examined. A release of copper from the Cu(I)-thiolate clusters of the protein by this remarkably important reagent was observed in vitro. The characteristic spectroscopic signals of the Cu(I)-thiolate chromophores levelled off in the presence of a two-fold molar excess of NO expressed per equivalent of thionein-copper as monitored by UV-electronic absorption, circular dichroism and luminescence emission. At the same time all of the copper became EPR detectable. The oxidized metal ions could easily be removed from the protein moiety by gelfiltration. The reversibility of the copper releasing process is of special interest. The specific fluorescence and dichroic properties of the previously demetallized protein could be recovered up to 85% under reductive conditions. Moreover, no difference in the electrophoretic behaviour was seen compared to the untreated Cu-MT. Thus, NO may act as a potent metabolic source for the transient copper release from Cu-MT. In the course of an oxidative burst this highly Fenton active copper is able to improve the efficacy of biological defence mechanisms.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aravindakumar CD, Ceulemans J, De Ley M. 1999 Nitric oxide induces Zn2+release from metallothionein by destroying zincsulphur clusters without concomitant formation of S-nitrosothiol. Biochem J344, 253–258.
Barnes PJ. 1995 Nitric oxide and airway disease. Ann Med27, 289–293.
Culotta VC, Lin SJ, Schmidt P, Klomp LW, Casareno RL, Gitlin J. 1999 Intracellular pathways of copper trafficking in yeast and humans. Adv Exp Med Biol448, 247–254.
Deters D, Hartmann H-J, Weser U. 1994 Transient thiyl radicals in yeast Cu(I)thionein. Biochim Biophys Acta1208, 344–347.
Felix K, Weser U. 1988 Release of copper from yeast copperthionein after S-alkylation of copper-thiolate clusters. Biochem J252, 577–581.
Felix K, Lengfelder E, Hartmann H-J, Weser U. 1993 A pulse radiolytic study on the reaction of hydroxyl and superoxide radicals with yeast Cu(I)-thionein. Biochim Biophys Acta1203, 104–108.
Hartmann H-J, Morpurgo L, Desideri A, Rotilio G, Weser U. 1983 Reconstitution of stellacyanin as a case of direct Cu(I) transfer between yeast copper-thionein and ‘blue’ copper apoproteins. FEBS Lett152, 94–96.
Hartmann H-J, Gärtner A, Weser U. 1984 Oxidation of Cu(I)-thionein by enzymatically generated H2O2. Hoppe-Seyler's Z Physiol Chem365, 1355–1359.
Hartmann H-J, Gärtner A, Weser U. 1985 Copper dependent control of the enzymic and phagocyte induced degradation of some biopolymers; a possible link to systemic inflammation. Clin Chim Acta152, 95–103.
Hartmann H-J, Schechinger T, Weser U. 1987 Cu(II) transfer into apo-Cu2Zn2_superoxide dismutase from Cu-thionein oxidized by activated leukocytes. Inorg Chim Acta136, 107–110.
Hartmann H-J, Schechinger T, Weser U. 1989 Copper-thionein in leukocytes. Biol Met2, 40–44.
Hartmann H-J, Li Y-J, Weser U. 1992 Analogous copper(I) coordination in metallothionein from yeast and the separate domains of the mammalian protein. BioMetals5, 187–191.
Hartmann H-J, Deters D, Weser U. 1996 Copper-release from yeast Cu(I)-thionein by hypothiocyanite (OSCN_). BioMetals9, 345–349.
Kelm M, Schrader J. 1988 Nitric oxide release from the isolated guinea-pig heart. Eur J Pharmacol155, 313–316.
Misra RR, Hochadel JF, Smith GT, Cook JC, Waalkes MP, Wink DA. 1996 Evidence that nitric oxide enhances cadmium toxicity by displacing the metal from metallothionein. Chem Res Toxicol9, 326–332.
Moncada S, Higgs A. 1993 The L-arginine-nitric oxide pathway. N Engl J Med329, 2002–2012.
Rae TD, Smith PJ Pufahl RA, Culotta VC, O'Halloran TV. Undetectable intracellular free copper: The requirement of a copper chaperone for superoxide dismutase. 1999 Science284, 805–808.
Richter A, Weser U. 1988 Kinetics of the H2O2 dependent cleavage of Cu-thiolate centres in yeast Cu8-thionein. Inorg Chim Acta151, 145–148.
Robbins RA, Grisham MB. 1997 Nitric oxide. Int J Biochem Cell Biol29, 857–860.
Schechinger T, Hartmann H-J, Weser U. Copper transport from Cu(I)-thionein into apo-caeruloplasmin mediated by activated leucocytes. Biochem J240, 281–283.
Snyder SH, Bredt DS. Biological roles of nitric oxide. Sci Am266, 68–77.
Traylor TG, Sharma VS. 1992 Why NO? Biochemistry31, 2847–2849.
Valentine JS, Gralla EB. 1997 Delivering copper inside yeast and human cells. Science278, 817–818.
Weser U, Hartmann H-J. 1984 Copper-thiolate proteins (metallothioneins). In: Lontie R. ed. Copper Proteins and Copper Enzymes, Vol. III Boca Raton, FL, CRC Press. 151–173.
Weser U, Mutter W, Hartmann H-J. 1986 The role of Cu(I)-thiolate clusters during the proteolysis of Cu-thionein. FEBS Lett197, 258–262.
Weser U, Hartmann H-J. Differently bound copper(I) in yeast Cu8-thionein. Biochim Biophys Acta953, 1–5.
Weser U, Hartmann H-J. 1991 Purification of yeast Cu-thionein. Methods Enzymol205, 274–278.
Zhang J, Snyder SH. 1995 Nitric oxide in the nervous system. Ann Rev Pharmacol Toxicol35, 213–233.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hartmann, HJ., Weser, U. Copper-release from yeast Cu(I)-metallothionein by nitric oxide (NO). Biometals 13, 153–156 (2000). https://doi.org/10.1023/A:1009275122084
Issue Date:
DOI: https://doi.org/10.1023/A:1009275122084