Studies on the gamma-glutamyl Cu-binding peptide from Schizosaccharomyces pombe.
The gamma-glutamyl peptide induced in Schizosaccharomyces pombe in response to metal stress has been purified following exposure of the organism to cadmium and copper salts. Induction of the peptide enables S. pombe to proliferate in media containing high concentrations of cadmium and copper. Two Cd-gamma-Glu peptide complexes are produced which differ in the content of acid-labile sulfur. One Cu-gamma-Glu peptide complex is induced, and it lacks acid-labile sulfur in the metal-binding cluster. The peptides are composed of repeating dipeptide units of gamma-Glu-Cys with a carboxyl-terminal glycine with heterogeneity observed in the repeat unit n. The number of repeats averages 3.2 and 3.8 for the Cd-peptides I and II and 3.6 for the Cu-peptide, in the case of the Cu-complex peptides with n values from 2 to 4 were separated by reverse phase high pressure liquid chromatography. The Cu-gamma-Glu peptide complex is oligomeric, but the exact number of peptide units per complex is not known. The copper binding stoichiometry averages 2.3 g atoms of Cu/mol of peptide, whereas Cd-peptides I and II average 1.8 and 2.7 mol eq of Cd(II)/peptide unit. The pH of half-dissociation of Cu ions from the gamma-Glu peptide is near 1.3, whereas pH values of 4 and 5.4 are sufficient for half-displacement of Cd ions from the sulfide-containing and -lacking peptides II and I, respectively. In the Cu-peptide complex copper is bound as Cu(I) as the complex exhibits luminescence characteristic of Cu(I)-S chelation. The luminescence emission peaks at 619 nm with a corrected excitation peak centered at 290 nm. The luminescence of the Cu-complex indicates the clustering of Cu(I) ions within a solvent-inaccessible complex. The complex is air-labile as the luminescence emission is gradually lost upon air exposure.
Reese
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Department of Medicine, University of Utah Medical Center, Salt Lake City 84132.
Mehra
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Tarbet
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Winge
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D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003300:Copper / Q000378:metabolism*; D008163:Luminescent Measurements; D009994:Osmolar Concentration; D010455:Peptides / Q000378:metabolism*; D004718:Saccharomycetales / Q000032:analysis*; D012568:Schizosaccharomyces / Q000032:analysis*; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D010455:Peptides; C053690:poly(gamma-glutamylcysteinyl)glycine; D003300:Copper