Release of copper from yeast copper-thionein after S-alkylation of copper-thiolate clusters.
Our knowledge on the release of copper from Cu-thionein in biological systems is limited. Other than oxidative cleavage or direct transfer, the possibility of an alkylation mechanism seemed attractive. Iodoacetamide and methyl methanesulphonate were successfully employed to alkylate the Cu-thiolate sulphur atom of homogeneous Cu(I)-thionein from yeast. The alkylation caused a weakening of the Cu-S bonding, which led to the release of copper. After equilibrium dialysis a proportion of the released copper was found in the dialysis buffer. When iodoacetamide was used carboxymethylcysteine was detected in the protein hydrolysate. A 10-fold molar excess over cysteine was sufficient for complete alkylation, which could be conveniently monitored by c.d. at 328 and 359 nm. The reaction proceeded under both aerobic and anaerobic conditions. E.p.r. measurements of Cu2+ revealed unequivocally the complete cleavage of the Cu-thiolate bonding in less than 5 h. It is possible that this mode of copper release might be of relevance to the molecular transport of this biochemically important transition metal.
D000478:Alkylation; D002352:Carrier Proteins; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D003300:Copper / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007460:Iodoacetamide / Q000494:pharmacology; D008668:Metallothionein / Q000378:metabolism*; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009942:Organometallic Compounds / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013696:Temperature
C500769:CUP1-1 protein, S cerevisiae; D002352:Carrier Proteins; D009942:Organometallic Compounds; C007121:copper thionein; C048402:copper(I)-thiolate; D003300:Copper; D008668:Metallothionein; D008741:Methyl Methanesulfonate; D007460:Iodoacetamide