Masaaki Hirobe

Base liberation from nucleotides by superoxide and intramolecular enhancement effect of phosphate group

The reaction of nucleotides with superoxide gave the corresponding nucleobases in good yield. The base-release reaction of nucleotides due to superoxide was examined and compared to that of nucleosides. Hence, the phosphate moiety greatly enhances the yields, in particular those of adenine from adenosine monophosphates. Superoxide in combination with the phosphate moiety has been revealed to be more active than superoxide alone. The phosphate peroxy radical, generated in situ from superoxide and phosphate, seems to be the species responsible for the formation of the free bases.

Novel Iron Complexes Behave Like Superoxide Dismutase In Vivo

Novel iron and copper complexes having tris[N-(5-methyl-2-pyridylmethyl)-2-aminoethyl]amine (5MeT-PAA), tris[N-(3-methyl-2-pyridylmethyl)-2-aminoethyl]amine (3MeTPAA), tris[N-(5-methoxycarbonyl-2-pyridylmethyl)-2-aminoethyl]amine (TNAA), tris[(2-thienylmethyl)-2-aminoethyl]amine (TTAA), tris[(2-furylmethyl)-2-aminoethyl]amine (TFAA) or tris[(2-imidazoyl)-2-aminoethyl]amine (TIAA) as ligand, were synthesized to examine the superoxide dismutase (SOD) activity. The concentrations of Fe-3MeTPAA and Fe-TIAA equivalent to 1 unit of SOD (IC50) were 0.5 microM and 1.0 microM, respectively. Fe-3MeTPAA and Fe-TIAA had higher SOD activity than other Fe and Cu complexes and protected Escherichia coli cells from paraquat toxicity. In case of using tris[N-(6-methyl-2-pyridylmethyl)-2-aminoethyl]amine (6MeTPAA) as ligand, the Fe complex could not be obtained, which may be due to the steric hindrance of 6-methyl substituent. Generally, Cu complexes had low SOD activity, compared with Fe complexes, and could not suppress paraquat toxicity.

Toxicity of 1-methyl-4-phenylpyridinium derivatives in Escherichia coli☆

Several derivatives of 1-methyl-4-phenylpyridinium (MPP+), i.e., 1-methyl-4-(4-nitrophenyl)pyridinium (1), 1-methyl-4-(4-cyanophenyl)pyridinium (2), 1-methyl-4-(3-nitrophenyl)pyridinium (3), 1-methyl-4-(4-chlorophenyl)pyridinium (4), 1-methyl-4-(4-acetamidophenyl)pyridinium (5), and 1-methyl-4-(4-aminophenyl)pyridinium (6), were synthesized in order to compare their toxicity with that of paraquat (PQ2+) in Escherichia coli. Addition of compounds 1, 2, and 3 to aerobic E. coli cell suspensions caused extracellular ferricytochrome c reduction, which was inhibited by superoxide dismutase in the same manner as that in the case of PQ2+. The rate of the ferricytochrome c (cyt. c) reduction was in the order of PQ2+ greater than 1 greater than 2 greater than 3, which is the same as that of the redox potentials of these compounds. On the other hand, MPP+, 4, 5, and 6, which have more negative potentials, had no effect on the cyt. c reduction. Compound 1 inhibited the growth of E. coli under aerobic conditions, but not under anaerobic conditions. The results show that compound 1 can act as a mediator for production of superoxide (O2-.), which seriously injures E. coli cells. However, though compounds 2 and 3 catalyzed the production of O2-. in E. coli cells, their activity of O2-. production was much lower than that of compound 1 or PQ2+. Thus, compound 3 had no effect on growth or survival of E. coli at 1 mM, while compounds 2 and 4 had both bacteriostatic and bacteriocidal effects which were independent of dioxygen (O2). The results show that the toxic mechanism is different from that of compound 1. MPP+, 5, and 6 had no effect on growth of E. coli. This paper shows that compound 1 is a novel enhancer of intracellular superoxide production, though the mechanism of toxicity of compounds 2 and 4 is not clear yet. The results suggest that the redox potential is a crucial factor for manifestation of the activity.

Dioxathiadiaza-heteropentalenes mediate superoxide and hydrogen peroxide production in Escherichia coli☆

The dioxathiadiaza-heteropentalenes, HEP-I (4,4-dimethyl-1,7-dioxa-2,6-diaza- 7 alpha lambda 4-thia-3H,5H-benzo[cd]pentalene), HEP-II (1,7-dioxa-2, 6-diaza-4, 7 alpha lambda 4-dithia-3H, 5H-benzo[cd]pentalene), HEP-III (1,7-dioxa-2,6-diaza-4, 7 alpha lambda 4-dithia-3H, 5H-benzo[cd]pentalene-4-oxide), and HEP-IV (1,7-dioxa-2,6-diaza-4,7 alpha lambda 4-dithia-3H, 5H-benzo[cd]pentalene-4,4-dioxide), inhibited growth of Escherichia coli in a simple glucose-salt medium, with their toxicities following the order of HEP-IV greater than HEP-III greater than HEP-II greater than HEP-I. These toxicities could be suppressed by yeast extract added to the glucose-salt medium. Yeast extract also facilitated maximal induction of superoxide dismutase (SOD) and catalase. The redox potentials of HEP-I-HEP-IV and the rates of oxygen uptake dependent on heteropentalenes in cyanide-resistant respiration of E. coli were correlated with the induction of SOD and catalase. Thus, the higher the redox potential of the compounds, the more potent they were for induction of enzyme production. Under anaerobic conditions, HEP-IV did not inhibit E. coli growth. These results indicate that HEP-I-HEP-IV can be reduced within the cell of E. coli and then reoxidized by molecular oxygen, generating O2- and H2O2. The toxicities of the heteropentalenes depend largely upon superoxide and/or hydrogen peroxide toxicity, and SOD and catalase provide a defense against the potential cytotoxicity of these species.