Effects of purine riboside on nucleic acid synthesis in ascites cells.

Purine riboside (nebularine, 9-beta-ribofuranosylpurine) is a naturally occurring base analog which closely resembles adenosine. It inhibits carcinogenic growth. Purine riboside strongly inhibits RNA and DNA synthesis in different cancer ascites cells. Gel electrophoretic analysis of RNA synthesis in vivo in the presence of purine riboside shows the ribosomal components to be inhibited the most. A method for assaying purine riboside or its phosphates intracellularly has been devised, and by using this it has been shown that purine riboside is extensively phosphorylated in the cells. The triphosphate derivative of purine riboside has been isolated and tested in the Escherichia coli RNA polymerase assay. It appears not to be incorporated into this type of RNA and to competitively inhibit this reaction with regard to ATP.     

Crystal structure of human nicotinamide riboside kinase

Nicotinamide riboside kinase (NRK) has an important role in the biosynthesis of NAD(+) as well as the activation of tiazofurin and other NR analogs for anticancer therapy. NRK belongs to the deoxynucleoside kinase and nucleoside monophosphate (NMP) kinase superfamily, although the degree of sequence conservation is very low. We report here the crystal structures of human NRK1 in a binary complex with the reaction product nicotinamide mononucleotide (NMN) at 1.5 A resolution and in a ternary complex with ADP and tiazofurin at 2.7 A resolution. The active site is located in a groove between the central parallel beta sheet core and the LID and NMP-binding domains. The hydroxyl groups on the ribose of NR are recognized by Asp56 and Arg129, and Asp36 is the general base of the enzyme. Mutation of residues in the active site can abolish the catalytic activity of the enzyme, confirming the structural observations.     

Confirmation of the anomeric structure of galacturonic acid in the galacturonosyl-ceramide of Sphingomonas yanoikuyae

The anomeric structure of glycosphingolipids significantly influences their activity to stimulate natural killer T cells. In this study the chemical structure of the galacturonosyl-ceramide in Sphingomonas yanoikuyae, designated GSL-1'sy, was re-examined to prove the anomeric structure of the Dgalacturonic acid (GalA) in the lipid, which was reported as beta-configuration by Naka et al., but was suggested as alpha-configuration in our preliminary study. GSL-1'sy was purified from the bacterial cells with the same procedure as Naka et al. The 1H-NMR analysis of GSL-1'sy revealed that the coupling constant of the anomeric proton of GalA was 3.0 Hz, indicating that GalA in GSL-1'sy is alpha-anomer, the configuration active for the stimulation of natural killer T cells.     

Confirmation of the structures of the phytoalexins lathodoratin and methyl-lathodoratin by synthesis

The two chromone phytoalexins were synthesized by condensing 2,4,6-trihydroxybutyrophenone with dimethylformamide.     

An efficient and scalable synthesis of 2,6-diaminopurine riboside

An efficient process to synthesize 2,6-diaminopurine riboside in high yield and quality is described. This inexpensive approach was scaled up to multi-hundred kilogram quantities for use in oligonucleotide therapeutics.     

Efficient synthesis of benzamide riboside, a potential anticancer agent

An efficient five step synthesis of benzamide riboside (BR) amenable for a large scale synthesis has been developed. It allows for extensive pre-clinical studies of BR as a potential anticancer agent.     

Effects of imidazoleacetic acid on brain amino acids and body temperature in mice

Free amino acids have been studied in the brains of fasted mice (18 h) injected intraperitoneally with a 3 mmol/kg dose of imidazole-4-acetic acid (IMA). Groups of mice were killed by cervical dislocation and their brains were removed before injection or at 5, 15, 30, 60, 90 and 120 min after injection and treated immediately with perchloric acid. Amino acid analyses were performed on the perchlorate extracts. Of the 16 amino acids evaluated quantitatively, only glutamic acid and glutamine showed progressive changes during the period of observation, the glutamic acid falling and glutamine levels rising. Serine and threonine levels were increased significantly above the control values from 30 min after the injection to the end of the experiment. Rectal temperatures (measured with a thermistor probe) after injection of IMA showed a progressive reduction from the control levels throughout the period of observation. An essentially linear correlation was noted between the decreases in body temperature and the differences between the glutamic acid and glutamine values for the first 90 min post-injection. Our data suggest that IMA affects mechanisms of temperature regulation, possibly in the hypothalamus, and that, among other processes, the activities of glutaminase and of serine and threonine dehydratases in brain might be reduced when brain temperatures fall.     

Confirmation of structure of the flavonol glucoside tambuletin

The structure of tambuletin, present in seeds of Xanthoxylum species has been confirmed as the 8-glucoside of gossypetin 7,4′-dimethyl ether.     

Metabolism of intracerebrally administered histidine, histamine and imidazoleacetic acid in mice and frogs

Abstract— After intracerebral administration of [¹⁴C]histidine to mice the major labelled substance found in the brain extracts was histidine itself; small amounts of labelled carnosine and homocarnosine were detected. No other labelled substances were detected on radio- autographs of two-dimensional TLC's of the extracts. In the case of the frog, radioactive histidine, N-acetylhistidine, carnosine and homocarnosine were found in the brain extracts at various times after intracerebral injection of the labelled histidine. With time, approximately 90 per cent of the radioactivity in the extracts was found in the N-acetylhistidine. In neither the mouse nor frog could we find unequivocal evidence for the formation either of histamine or imidazoleacetic acid from intracerebrally administered histidine, but our analytical procedures may have lacked sufficient sensitivity to pick up extremely low activities of histamine and imidazoleacetic acid. Experiments with [¹⁴C]histamine administered intracerebrally into mice demonstrated the major pathway of metabolism in brain to be histamine → methylhistamine → methylimidazoleacetic acid. No detectable label appeared in inlidazoleacetic acid. In the frog intracerebral administration of the labelled histamine led to the formation of methylhistamine and imidazoleacetic acid, but at most only traces of methylimidazoleacetic acid were found. The injection of [¹⁴C]imidazoleacetic acid intra- cerebrally into mice and frogs resulted in virtually no loss of the label in the form administered in the frog brain over a period of 4 h and in a slow rate of decrease in the mouse brain. No radioactive metabolites of imidazoleacetic acid were found in either species. The limitations of trying to determine natural functions of substances in brain by following the fate of exogenously administered materials is discussed.     

Ribosyl-diphthamide: Confirmation of structure by fast atom bombardment mass spectrometry

Diphtheria toxin inactivates protein synthesis elongation factor 2 by attaching ADP-ribose to an unusual post-translational amino acid derivative, diphthamide, in the factor. Previously, we prepared ribosyl-diphthamide from the ADP-ribosyl-factor and proposed on the basis of NMR spectral analysis that it is 1-α-d-ribofuranosyl-2-[3-carboxyamido-3-(trimethylammonio)propyl]histidine [N. J. Oppenheimer, and J. W. Bodley, (1981) J. Biol. Chem.256, 8579–8581 and op. cit.]. Now, using fast atom bomardment mass spectrometry, the intact cation of ribosyl-diphthamide has been observed in the gas phase. The theoretical mass of the structure proposed for ribosyl-diphthamide uniquely agrees with the observed mass of the inact cation of the compound to within 2 ppm. Collisional activation decomposition mass spectral analysis provided additional structural confirmation. Thus, although the compound has not been synthesized, all available evidence appears uniquely consistent with the structure of ribosyl-diphthamide previously proposed.     

Confirmation of primitive chromosome structure in the hexaploid wheats

Summary Hybrids between Triticum aestivum cv Chinese Spring (CS) and T. dicoccoides of chromosome type E_1a showed only a few or no trivalents at meiosis, but both trivalents and quadrivalents were shown by hybrids with six other types. Since there is strong evidence that the E_1a type has the primitive chromosome structure of T. dicoccoides, Chinese Spring can be said to have the primitive chromosome structure of the hexaploid wheats in regard to reciprocal translocation.Contribution no. 51 from the Plant Germ-plasm Institute, Faculty of Agriculture, Kyoto University, Kyoto, Japan