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T T Sakai

Publications and source records attributed to T T Sakai.

36 records · Page 2Linked to original sources
PubMed1981

Bleomycin-induced life prolongation of mice infected with Trypanosoma brucei brucei EATRO 110.

The antitumour antibiotic bleomycin, supplied as commercial BlenoxaneR (a mixture of bleomycinic acids), at 7 or 14 mg/kg prolonged life greater than 30 days beyond death of controls without relapse or sign of trypanosomes in the peripheral blood of mice infected with Trypanosoma b. brucei EATRO 110. Control mice died in three to four days. The purified A2 and B2 bleomycin congeners were also active at this dose. Spermidine, spermine and hirudonine (1,8-diamidinospermidine) but not putrescine, co-administered with drug, annulled the curative effects of these compounds, as signalled by appearance of trypanosomes in the blood and death of the animals.

Animals↗
PubMed1981

Synthesis and DNA binding of [3-[2'-(2-acetamidoethyl)-2,4'-bithiazole-4-carboxamido]propyl[dimethylsulfonium chloride, a fragment of bleomycin A2.

[3-[2'-(2-Acetamidoethyl)-2,4'-bithiazole-4-carboxamido]propyl]dimethylsulfonium chloride (1), the acetyl derivative of the cationic terminal dipeptide of bleomycin A2, has been synthesized and its binding to DNA and poly(dA-dT) has been studied by proton NMR and fluorescence spectroscopy. The spectral perturbations which occur upon binding of the compound to either nucleic acid indicate that that portion of bleomycin which binds to the nucleic acid can, for the most part, be mimicked by the fragment. The data are discussed in terms of the structure of the drug and the drug-nucleic acid complex.

Binding Sites↗
PubMed1981

Proton NMR studies of the Zn(II)--bleomycin-A2--poly(dA-dT) ternary complex.

Proton NMR spectra demonstrate the formation of a ternary complex, Zn(II)--bleomycin-A2--poly(dA-dT), which serves as an analog of the putative active complex, Fe(II)--bleomycin-A2--DNA. Specific sites of metal--drug and drug--nucleic acid interaction have been delineated on the basis of chemical shift perturbations. On the basis of this criterion there appear to be three distinct regions of the drug: (i) the NH2 terminus up to and including the disaccharide and hydroxyhistidine residues, whose resonances are perturbed only by metal interactions; (ii) the COOH-terminal dipeptide, whose resonances are displaced only by nucleic acid interactions; and (iii) the methylvaleric acid-threonine dipeptide, which links these domains and whose resonances are sensitive to both types of interactions. The spectral perturbations in the first and second domains are very similar to changes observed in the corresponding binary complexes--i.e., Zn(II)--bleomycin-A2 and bleomycin-A2--poly(dA-dT), respectively.

Bleomycin↗
PubMed1980

Carbon-13 N.M.R. study of bleomycin-A2 protonation.

The acid-base titration of bleomycin-A2 in D2O solution at 35 +/- 5 degrees has been monitored by 13C n.m.r. spectroscopy at 67.89 MHz. The following pKDa values were obtained: 3.68 +/- 0.05 (secondary amine), 5.29 +/- 0.03 (imidazole), and 8.23 +/- 0.19 (primary amine), where KDa is the dissociation constant in D2O solution. The equilibrium isotope effects (pKDa--pKa in H2O) are: 0.70 +/- 0.06 (secondary amine), 0.28 +/- 0.04 (imidazole), and 0.85 +/- 0.19 (primary amine). Titration of the imidazole group of Bleo-A2 occurs at Npi, i.e. only Ntau is protonated in basic solution. Significant protonation shifts are almost completely limited to carbons of the N-terminal tetrapeptide, suggesting that the C-terminal tripeptide extends into the solvent and interacts to a minimal extent with the rest of the molecule. Long range protonation shifts associated with titration of the imidazole and secondary amine groups indicate that protonation of one or both of these sites is probably accompanied by significant conformational changes. The observed protonation shifts generally fail to correlate with Zn(II) complexation shifts reported by Dabrowiak et al. (1973, Biochemistry 17., 4090) indicating that ligation sites cannot unambiguously be determined from these complexation shifts. The complexation shifts previously attributed to coordination of the imidazole and carbamoyl groups probably result from conformational changes.

Amines↗
PubMed1976

Regulation of ornithine decarboxylase activity by guanine nucleotides: in vivo test in potassium-depleted Escherichia coli.

The observation that guanosine 5'-triphosphate (GTP) is an activator and guanosine 5'-diphosphate-3'-diphosphate (ppGpp) is an inhibitor of ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) of Escherichia coli [E. Hölttä et al. (1974) Biochem. Biophys. Res. Commun. 59, 1104-1111] has been confirmed. The hypothesis that synthesis of both polyamine and RNA in E. coli is regulated in vivo by these nucleotides was tested in E. coli B-207. On transfer of this K+-requiring, amino-acid-deficient strigent strain from K+-medium to Na+-medium, the organism stops protein synthesis, maintains a high rate of RNA synthesis, and increases putrescine synthesis from ornithine manyfold. Under these conditions, the cells do not markedly change their contents of GTP and ppGpp. The proposed mechanism of regulation of RNA and putrescine synthesis by guanine nucleotides does not appear to explain the metabolic phenomena observed in this organism during K+ deficiency. Nevertheless, amino acid depletion in K+-medium does result in a marked increase in ppGpp.

Adenosine Triphosphate↗
PubMed1975

The binding of polyamines and of ethidium bromide to tRNA.

The binding of spermidine and ethidium bromide to mixed tRNA and phenylalanine tRNA has been studied under equilibrium conditions. The numbers and classes of binding sites obtained have been compared to those found in complexes isolated by gel filtration a low ionic strength. The latter complexes contain 10-11 moles of either spermidine or ethidium per mole of tRNA; either cation is completely displaceable by the other. In ethidium complexes, the first 2-3 moles are bound in fluorescent binding sites; the remaining 7-8 molecules bind in non-fluorescent form. At least one of the binding sites for spermidine appears similar to a binding site for fluorescent ethidium. Similar results are found with E. coli formylmethionine tRNA. Spermine, in excess of 18-20 moles per mole tRNA, causes precipitation of the complex. Putrescine does not form isolable complexes with yeast tRNA and displaces ethidium less readily from preformed ethidium-tRNA complexes. Under equilibrium conditions, in the absence of Mg++, there are 16-17 moles of spermidine bound per mole of tRNA as determined by equilibrium dialysis. Of these, 2-3 bind with a Ksence of 9 mM Mg++, the total number of binding sites is decreased slightly and there appears to be only one class of sites with a Ka = 600 M(-1). Quantitatively similar results are obtained for the binding of spermidine to yeast phenylalanine tRNA. When the interaction between ethidium bromide and mixed tRNA is studied by equilibrium dialysis or spectrophotometric titration, two classes of binding sites are obtained: 2-3 molecules bind with an average Ka = 6.6 x 10(5) M(-1) and 14-15 molecules bind with an average Ka = 4.1 x 10(4) M(-1). Spermidine, spermine, and Mg++ compete effectively for both classes of ethidium sites and have the effect of reducing the apparent binding constants for ethidium. When the binding of ethidium is studied by fluorometry, there are 3-4 highly fluorescent sites per tRNA. These sites are also affected by spermidine, spermine and Mg++. Putrescine has little effect on any of the classes of binding sites. These data are consistent with those found under non-equilibrium conditions. They suggest that polyamines bind to fairly specific regions of tRNA and may be involved in the maintenance of certain structural features of tRNA.

Binding Sites↗
PubMed1975

Interrelation between guanosine tetraphosphate accumulation, ribonucleic acid synthesis, and streptomycin lethality in Escherichia coli CP78.

The effects of streptomycin on the synthesis of ribonucleic acid (RNA) and guanosine-3'-diphosphate-5'-diphosphate have been examined in the isogenic strains CP78 (rel(+)) and CP79 (rel-). In the absence of the required amino acids arginine, leucine, or threonine, streptomycin stimulates RNA synthesis in CP78 and this stimulation coincides with cell death. However, in the absence of histidine, also a required amino acid, streptomycin kills the cells without stimulation of RNA synthesis above that which occurs in the absence of streptomycin. In all of these instances, guanosine-3'-diphosphate-5'-diphosphate levels vary inversely with RNA synthesis, decreasing when a stimulation of RNA synthesis occurs. Streptomycin has little effect on guanosine-3'-diphosphate-5'-diphosphate levels in CP78 in the absence of histidine. Another histidine auxotroph, 15 T(-)H(-)U(-) (rel(+)), does show streptomycin-stimulated synthesis of RNA which is coincident with cell death. CP79 (rel(-)) is 10 times more susceptible to streptomycin than CP78 and streptomycin causes an inhibition of the relaxed synthesis of RNA. The greater susceptibility of CP79 may be due to a greater initial uptake of streptomycin by CP79.

Escherichia coli↗