The role of Cu(I)-thiolate clusters during the proteolysis of Cu-thionein.
Rat liver Cu,Zn-[35S]thionein and yeast Cu-thionein were subjected to proteolysis in vitro using equilibrium dialysis. The partially copper-loaded vertebrate thionein (2-7 Cu/mol) was affected by different proteases including thermolysin, proteinase K, protease from Streptomyces griseus and lysosomal enzymes. Unlike the 2Cu-thionein the respective 7Cu-thiolate-centred metallothionein was hardly proteolytically digested. In contrast to fully copper-loaded native yeast Cu-thionein both the H2O2-oxidized and the metal-free protein were effectively cleaved in the presence of proteinase K. It is important to realize that the native Cu(I)-thiolate chromophore survives the proteolytic attack. When the copper-sulphur bonding is broken and the same amount of copper is unspecifically bound to the thionein portion, proteolysis proceeds identically with respect to the rate observed in the presence of the apoprotein. The unsuccessful proteolysis of native Cu-thionein is not attributable to a simple copper-dependent inhibition of the proteinases. It is suggested that prior to proteolysis the copper-sulphur clusters must be destroyed.
Weser
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Mutter
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Hartmann
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D000818:Animals; D002352:Carrier Proteins; D003300:Copper / Q000378:metabolism*; D019286:Endopeptidase K; D010450:Endopeptidases / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008668:Metallothionein / Q000378:metabolism*; D009942:Organometallic Compounds / Q000378:metabolism*; D010084:Oxidation-Reduction; D010447:Peptide Hydrolases / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013305:Streptomyces griseus / Q000201:enzymology; D013820:Thermolysin / Q000378:metabolism
C500769:CUP1-1 protein, S cerevisiae; D002352:Carrier Proteins; D009942:Organometallic Compounds; C007121:copper thionein; C048402:copper(I)-thiolate; D003300:Copper; D008668:Metallothionein; D010450:Endopeptidases; D010447:Peptide Hydrolases; D019286:Endopeptidase K; D013820:Thermolysin