ascorbic-acid and diglycyl-histidine

Free divalent ions of copper (Cu) are capable of generating radical species such as hydroxyl radicals in the presence of hydrogen peroxide or ascorbic acid through Harbor-Weiss-like reactions under physiological conditions. It has been reported that radical-mediated damage to DNA molecules in animal cells leads to programmed cell death. Hence it is important to seek for methods to prevent Cu-mediated DNA damage. In this study we identified on effect of Cu binding of short peptides (chiefly Gly-Gly-His tripeptide) in the prevention of DNA degradation caused by Cu-mediated reactions in the presence of hydrogen peroxide and of ascorbic acid.

Topics: Ascorbic Acid; Carrier Proteins; DNA; DNA Damage; Hydrogen Peroxide; Hydroxyl Radical; Oligopeptides; Oxidation-Reduction; Reactive Oxygen Species

Glycylglycylhistidine (GGH) was synthesized using mixed anhydride method, and Cu(II): GGH = 1:1 complex was identified using UV, ESR, CD, ORD and FAB/MS technique. The oxidation of ascorbate catalyzed by cupric ion in RPMI 1640 medium (pH 7.40) was studied. The activation energy E = 39.2 kJ/mol. The addition of GGH inhibited the oxidation of ascorbate catalyzed by cupric ion. The investigations with ESR and fluorimetry indicate that there are Cu(I), ascorbate radical (AH), O2-., H2O2 and .OH during the oxidation.

Topics: Animals; Ascitic Fluid; Ascorbic Acid; Carcinoma, Ehrlich Tumor; Copper; Mice; Oligopeptides; Oxidation-Reduction

The DNA-cleavage specificity of ascorbate in the presence of copper ion is analyzed with end-labeled pBR322 DNA fragments. The nonenzymatic reaction of Cu(II)/ascorbate and DNA shows certain degrees of cleavage preference toward purine-containing short segments in the labeled DNA under mild conditions (at 0 degrees C and 10 min). The segments of pyrimidine clusters are least susceptible to cleavage. The DNA scission cannot be detected in the absence of metal ions, and is greatly diminished in the presence of EDTA and metal-chelating peptide. It is more specific than the nuclease-like scission activity induced by cuprous-phenanthroline complex. This scission activity in relation to the antiviral and antitumor activities of vitamin C reported in the literature deserves a crucial consideration.

Topics: Ascorbic Acid; Base Sequence; Chelating Agents; Copper; DNA, Bacterial; Edetic Acid; Oligopeptides; Oxidation-Reduction; Phenanthrolines; Plasmids

Ascorbate in an aqueous solution is easily oxidized by molecular oxygen in the presence of cupric ion, thus producing reactive oxygen species and exhibiting cytotoxicity. In order to increase the antitumor activity of ascorbate, we used the innocuous form of cupric ion complexed with glycylglycylhistidine, a tripeptide designed to mimic the specific Cu(II) transport site of albumin molecule. Although this square planar copper:glycylglycylhistidine complex did not significantly oxidize ascorbate at pH 7.4, it killed Ehrlich ascites tumor cells in vitro in a high concentration of ascorbate. The injections of large doses of ascorbate together with copper: glycylglycylhistidine prolonged the life span of mice inoculated i.p. with Ehrlich tumor cells. The target specificity against tumor cells was primarily attributable to their high peptide-cleaving activity.

Topics: Animals; Ascorbic Acid; Carcinoma, Ehrlich Tumor; Cell Survival; Copper; Drug Synergism; Electron Spin Resonance Spectroscopy; Kinetics; Mice; Oligopeptides; Oxidation-Reduction

L-Ascorbic acid, when combined with either copper(II) ion or a copper(II)-tripeptide complex, extensively cleaved several viral DNAs and proteins under in vitro conditions. Neither ascorbate nor copper tripeptide (Cu2+-diglycyl-L-histidine) alone caused any apparent changes on these molecules. Various transition metal ions and reducing agents were examined under comparable conditions to determine the basic requirements for both DNA degradation and protein scission activities. Copper and iron are the two most effective transition metal ions examined that exhibit these activities in the presence of ascorbate. The addition of catalase, but not superoxide dismutase, can partially inhibit the scission of DNA in vitro, suggesting that H2O2 may be involved in these activities. Among the various reducing agents tested, ascorbate was most effective in causing DNA scission and protein cleavage, corroborating the possible role of H2O2 in the cleavage reactions. One of the products of the reactions of copper/ascorbate is probably the hydroxyl radical generated from H2O2, which can be formed from the oxidation of ascorbate.

Topics: Ascorbic Acid; Chelating Agents; Copper; DNA; DNA, Viral; Humans; Metals; Oligopeptides; Proteins; Serum Albumin; Transferrin