Method for producing hypo-metallated redox-active metallothionein protein and pharmaceutical composition containing the same
AssigneesChristoph Melcher, Tony Raoul Henry, Chun-Hung Kuo
InventorsCHRISTOPH MELCHER, TONY RAOUL HENRY
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Abstract
The present invention relates to method for producing hypo-metallated redox-active metallothionein (MT) proteins, pharmaceutical compositions containing the proteins, and uses the pharmaceutical compositions for treatment of conditions originating from elevated intracellular oxidative stress and/or dis-balanced intracellular redox-potential and/or redox-potential-dependent imbalance of metal ions.
Claims16
A method for producing a hypo-metallated redox-active metallothionein protein, the hypo-metallated redox-active metallothionein protein having 20 cysteine sulfhydryl groups and 7 binding pockets, 2 to 16 of the 20 cysteine sulfhydryl groups being free and reduced, and 1 to 6 of the 7 binding pockets being occupied by metal ions, comprising:
- producing a metallothionein protein;
- purifying the metallothionein protein by utilizing cleavage of histidine tag and metal affinity chromatography;
- de-metallating the metallothionein protein;
- chemically reducing the de-metallated metallothionein protein;
- and
- partially metallating the reduced de-metallated metallothionein protein with 4 molar equivalents of zinc ions,
- wherein the purified metallothionein protein having amino acid sequence of SEQ ID NO:
- 6.
Dependencies are inferred from the claim text.