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Biomedical subjects

D Shugar

Publications and source records attributed to D Shugar.

At least 109 records · Page 6Linked to original sources
PubMed1981

Comparison of solid state and solution conformations of R and S epimers of 8,5'-cycloadenosine and their relevance to some enzymatic reactions.

The C(5')-R epimer of 8,5'-cycloadenosine crystallizes in the monoclonic space group P2(1) (Z = 2) with unit cell dimensions a = 5.755 (1), b = 16.895 (1), and c = 5.511 (1) A and beta = 104.16 (1) degree. X-ray intensity data were measured on a diffractometer, and the crystal structure was determined by direct methods. Least-squares refinement converged at R = 0.037 for 1008 reflections. The conformation about the glycosyl bond is anti, as imposed by the 8,5' cyclization, with XCN = 29.8 degrees. The ribose ring adopts the unusual C(1')endo-O(4')exp (o1T) conformation with pseudo-rotation parameters P = 289.0 degrees and tau m = 490 degrees. The six-membered ring formed by the 8,5' linkage is approximately a half-chair with C(4') and O(4'), respectively, above and below a plane defined by the other four atoms. These results are compared with those previously reported for the corresponding S epimer [Haromy, T. P., Raleigh, J., & Sundaralingam, M. (1980) Biochemistry 19, 1718-1722]. The conformations of the sugar rings and the exocyclic groups of both epimers in the solid state are compared to the conformations in solution, as determined by analysis of the systems of proton-proton vicinal coupling constants from the 1H NMR spectra. The foregoing findings are employed to examine the role of the conformational parameters of adenosine and 5'-AMP in reactions catalyzed by the appropriate enzymes.

AMP Deaminase↗
PubMed1981

Phi W-14 DNA inhibits transfection of Bacillus subtilis by SPP1 DNA.

The DNA of bacteriophage phi W-14 is unusual in that half of the thymine residues are replaced with the hypermodified pyrimidine alpha-putrescinylthymine (Kropinski et al., Biochemistry 12:151-157, 1973). Bacteriophage phi W-14 DNA and Bacillus subtilis DNA exhibited comparable competing abilities for the uptake of transfecting bacteriophage SPP1 DNA by competent cells of B. subtilis. B. subtilis DNA decreased transfection and uptake to the same extent, indicating that it merely competed with SPP1 DNA for uptake. Phi W-14 DNA, however, decreased transfection up to 30 times more effectively than it inhibited uptake. Phi W-14 DNA did not alter the kinetics of transfection. The degree of inhibition of transfection was dependent upon the time of addition of Phi W-14 DNA relative to the time of addition of SPP1 DNA. If failed to inhibit when added 30 min after SPP1 DNA. It had a fourfold-greater effect when added 10 min before, rather than simultaneously with, SPP1, but this enhancement was abolished by high concentrations of SPP1 DNA. The nature of the transfection process was not altered in those cells escaping inhibition by Phi W-14 DNA: two molecules of transfecting SPP1 DNA were required to form a transfectant with or without Phi W-14 DNA. Free putrescine did not affect transfection by SPP1 DNA. It was concluded that the putrescine groups covalently attached to phi W-14 DNA allowed this DNA to interfere with the transfection process at the intracellular level.

Bacillus subtilis↗
PubMed1981

Pyrimidine arabinofuranosyl nucleosides with 5-substituted long, branched and unsaturated chains: synthesis and antiherpes properties.

Several procedures have been applied to the preparation of 5-alkyl analogues of araU and araC via condensation of the appropriate 2,4-bis-O-(trimethylsilyl)-5-alkylpyrimidine with 2,3,5-tri-O-benzyl-alpha-D-arabinofuranosyl chloride. The resulting O'-benzylated nucleosides were deblocked with the aid of BF3 . Et2O in C2H5SH. The araC analogues were also prepared by conversions of the corresponding 5-alkyl-araU derivatives. The chloromercuri derivatives of araU and araC, were reacted with allyl chloride in the presence of Li2PdCl4. The resulting 5-allyl derivatives were catalytically reduced to the corresponding 5-propyl analogues. Catalysed condensation of 2,4-bis-O-(trimethylsilyl)-5-vinyluracil with 2,3,5-tri-O-benzyl-alpha-D-arabinosyl chloride, as well as with 1-O-acetyl-2,3,5-tri-O-benzoylarabinofuranose, was carried out under a variety of different conditions. Deblocking of the benzylated nucleosides with various reagents led invariably to addition to the vinyl substituent. In the case of benzoylated nucleosides, deblocking yielded largely the alpha-anomers. The antiherpes activities of the 5-alkyl compounds have been evaluated, as well as the susceptibility of the araC analogues to enzymatic deamination.

Antiviral Agents↗
PubMed1980

Comparison of theoretical and experimental approaches to determination of conformation of nucleosides about the glycosidic bond.

A study has been made by means of 1H-NMR spectroscopy of the syn in equilibrium anti dynamic equilibrium about the glycosidic bond for 5'-deoxyadenosine and some 8-substituted analogues, in different solvents. The results are compared with those previously obtained for the parent adenosine and its 8-substituted analogues. Quantum chemical calculations, with the aid of the Classical Potential and PCILO procedures, were applied to obtain the energies for different conformations of the base in adenosine and 5'-deoxyadenosine, and their 8-methyl and 8-halogeno derivatives. Good agreement was found between experimentally determined conformations in solution and those corresponding theoretically to the energy minima, particularly those calculated by the PCILO method. Comparison of the quantitative experimental data with the theoretical results was used to evaluate the validity of the latter and their applicability to studies of nucleoside conformation. The experimental and theoretical findings pointed to the existence of a marked flexibility about the glycosidic bond of the parent nucleosides and their 8-substituted analogues, when the 8-substituents were not too bulky, such as methyl or bromine. Considerations is given to possible correlations between conformational parameters in nucleosides and their 5'-deoxy analogues. It is shown that the proposed stabilization of the conformation syn by intramolecular hydrogen bonding, 5'-OH...N(3), is not in accord with the results of the present study.

Adenosine↗
PubMed1980

Properties of purified salivary ribonuclease, and salivary ribonuclease levels in children with cystic fibrosis and in heterozygous carriers.

Alkaline acid- and thermo-stable ribonuclease was isolated from human saliva of healthy subjects, children with cystic fibrosis, and from parents of the latter. The enzyme was purified about 180-200 times, and its ionic requirements, and specificity towards various substrates, characterized and compared with those for human pancreatic ribonuclease. The level of acid- and thermo-stable ribonuclease activity was five times higher in terms of enzyme level per ml saliva, and three times higher in terms of salivary protein, for 62% of children with cystic fibrosis, and for 73% of carrier subjects, relative to co-nrol groups. Assay of the level of this ribonuclease activity appears to be a promising diagnostic tool for detection of heterozygote carriers.

Adult↗
PubMed1980

The effects of tetraalkylammonium salts on helix-coil transition parameters in natural and synthetic ribo- and deoxyribo-polynucleotides.

Thermal transition profiles were recorded for a variety of natural and synthetic DNA and double-stranded RNA preparations in the presence of tetramethylammonium (TMA+) and tetraethylammonium (TEA+) cations. Double-stranded RNAs of natural origin, with GC contents of 50% exhibited the same profiles and Tm values as native DNA containing normal bases. Hence the tetraalkylammonium cations liquidate not only the effects of base composition, and the difference in stability between A-T and A-U base pairs (further confirmed by measurements with uracil-containing DNA from phage PBS-2), but also that of the 2'OH. In the presence of TMA+ cations, there is very marked enhancement of the stability of U-U base pairs in poly(rU) and poly(Um). In 2.4 M TEA, the 1:1 complex of poly(G) with poly (C) formed readily and melted reversibly with a Tm as low as 87 degrees C. At concentrations of TMA and TEA for which dTm/dXGC = 0, the Tm values for various phage DNA preparations containing atypical bases (phages T2, T4, phi e, phi W-14, PBS-2) differ appreciably from those with 'normal bases'. Analysis of these findings indicates that the selective interaction of TMA and TEA cations with A-T base pairs occurs in the minor groove of the DNA helix. The overall results show that the action of these quaternary ammonium cations is not due exclusively to preferential binding to A-T base pairs, but must involve other factors, including modifications of solvent structure. They also underline the utility of TMA and TEA solvent systems for placing in evidence transition profiles not accessible in other solvent systems.

DNA↗
PubMed1980

Comparative efficacy of antiherpes drugs against different strains of herpes simplex virus.

A large variety of antiherpes compounds was compared for their inhibitory activity against laboratory strains and clinical isolates of herpes simplex virus (HSV) type 1 and type 2. From studies performed in primary rabbit kidney cell cultures, six, E-5-(2-bromovinyl)-2'-deoxyuridine, E-5-(2-iodovinyl)-2'-deoxyuridine, 5-vinyl-2'-deoxyuridine, 2'-fluoro-5-iodoaracytosine, acycloguanosine, and 5-iodo-2'-deoxycytidine, emerged as the most potent and selective antiherpes agents. For HSV type 1, the 50% inhibitory doses (ID50) were 0.008, 0.012, 0.018, 0.017, 0.04, and 0.06 micrograms/ml, respectively; those for HSV type 2 were 1, 2, 0.1, 0.05, 0.04, and 0.3 microgram/ml, respectively. These compounds did not inhibit host-cell metabolism or replication of vaccinia virus except at concentrations 100--10,000 times greater than the ID50 for any HSV. All were significantly less inhibitory for a thymidine kinase (TK)-deficient mutant of HSV type 1 than for normal strains, suggesting that phosphorylation by virus-induced TK was required to produce specific inhibition of HSV replication.

Antiviral Agents↗
PubMed1980

NMR studies in the syn-anti dynamic equilibrium in purine nucleosides and nucleotides.

The syn in equilibrium anti equilibrium conformation about the glycosidic bond of purine nucleosides and 5'-nucleotides in different solvent systems has been investigated by means of 1H NMR spectroscopy. Quantitative values for the conformer populations were improved, relative to previous results, by a detailed study of, and a resultant derived correction for, the influence of the sugar exocyclic group conformation on the chemical shifts of the sugar ring protons. This was achieved with the aid of nucleosides and nucleotides fixed in the conformations gauche-trans [derivatives of 8,5'-(R)-cyclo] and trans-gauche [derivatives of 8,5'-(S)-cyclo]. The results of 13C NMR confirmed those obtained by 1H NMR. The measured values of the vicinal coupling constants between H-1' and the C-8 and C-4 carbons were employed to evaluate approximately the glycosidic angles chi of the nucleosides in the conformations syn and anti. A critical examination is made of the applicability of relaxation methods, involving analysis of spin-lattice relaxation time of protons (T1) and the Overhauser effect, to determine the conformation of the base about the glycosidic bond; interpretations are provided for the lack of agreement between these methods and those based on chemical shifts in the present study. The foregoing resuls are also applied to an examination of the effect of the conformation of the base about the glycosidic bond on the enzymatic reactions catalyzed by 5'-nucleotidase and adenosine deaminase.

Adenosine↗
PubMed1980

Influence of bacteriophage PBS1 and phi W-14 deoxyribonucleic acids on homologous deoxyribonucleic acid uptake and transformation in competent Bacillus subtilis.

Both bacteriophage PBS1 deoxyribonucleic acid (DNA) (in which all the thymine residues are replaced by uracil) and phage phiW-14 DNA [in which half the thymine residues are replaced by 5-(aminobutylaminomethyl)uracil or 5-putrescinylthymine] exhibit comparable competing abilities for uptake of homologous DNA in a Bacillus subtilis competent system. But, whereas PBS1 DNA leads to a decrease in transformation frequencies compatible with its competing ability for DNA uptake, phiW-14 DNA decreases transformation frequencies by a factor up to eightfold higher. The effect of phiW-14 DNA on transformation frequencies is visible even at a concentration level that does not decrease transforming DNA uptake. No such effect was observed with heterologous DNA containing presumably ionically bound putrescine. Low concentrations of phiW-14 DNA decreased the number of double (nonlinked) transformants more than single transformants. The influence on transformation was abolished when phiW-14 DNA was added 20 min after addition of transforming DNA, i.e., when the recombination process was terminated. The putrescine-containing DNA also decreased retention of trichloroacetic acid-precipitable radioactivity of homologous DNA taken up. We conclude that phiW-14 DNA inhibits some intracellular process(es) at the level of recombination. In addition, there is evidence that phiW-14 DNA, but not heterologous DNA with ionically bound putrescine, binds also to site(s) on the cell surface other than receptors for homologous DNA.

Bacillus subtilis↗
PubMed1980

Analogues of formycins A and B: synthesis and some properties of methyl derivatives of 7-amino and 7-keto pyrazolo(4,3-d)pyrimidines.

Starting from the synthesis of 3-propyl-7-ketopyrazolo(4,3-d)pyrimidine (a formal analogue of formycin B), this was converted to the 7-thio derivative, which was aminated to give 3-propyl-7-aminopyrazolo(4,3-d(pyrimidine (a formal analogue of formycin A) and its methylamino derivatives. With the aid of different methylating agents, all four possible ring N-mono-methyl derivatives of the foregoing have been synthesized. Dimethyl derivatives were prepared on a small scale and identified by cross-methylation reactions, spectral properties and chromatographic behaviour. The basic physico-chemical properties of all the foregoing are described, including ultraviolet absorption spectra, spectrophotometrically determined pK values, and the 1H chemical shifts in aqueous medium and/or dimthjyl sulphoxide. The spectral properties of the various analogues indicate that the N6-methyl derivatives exist predominantly in the imino form, in contrast to the predominant amino form of the corresponding analogue from the adenine series, N1-methyladenine, but similar to the fixed imino form of N1-methyl-9-substituted adenines.

Amination↗
PubMed1980

Mechanism of hydroxylamine mutagenesis: an infrared study of the association in non-polar solutions of 5-methyl-N4-hydroxycytosines.

An infrared spectroscopic study has been made of the structure and self-association of 1,5-dimethyl-N4-hydroxycytosine and 1,5-dimethyl-N4-methoxycytosine in several non-polar solvents as a function of concentration and temperature. Both compounds are predominantly in the imino form, and the overall results point to the existence of an intramolecular hydrogen bond between the ring N(3) hydrogen and the oxygen on the exocyclic N4, as previously found in the solid state. Furthermore, 1,5-dimethy-N4-hydroxycytosine forms linear associates in solution via hydrogen bonding, of the form N4-OH ... O=C2, with thermodynamic parameters delta H = -2.8 kcal/mol, delta S = -2.7 cal/deg . mol. As anticipated, the N4-methoxy derivatives does not form such associates. Neither of the compounds exhibits detectable association by hydrogen bonding with the potentially complementary 9-ethytl-adenine. Both compounds are also predominantly in the imino form in the gas phase.

5-Methylcytosine↗