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91.
NMR studies of multiple conformations in complexes of Lactobacillus casei dihydrofolate reductase with analogues of pyrimethamine 总被引:2,自引:0,他引:2
B Birdsall S J Tendler J R Arnold J Feeney R J Griffin M D Carr J A Thomas G C Roberts M F Stevens 《Biochemistry》1990,29(41):9660-9667
1H and 19F NMR signals from bound ligands have been assigned in one- and two-dimensional NMR spectra of complexes of Lactobacillus casei dihydrofolate reductase with various pyrimethamine analogues (including pyrimethamine [1, 2,4-diamino-5-(4'-chlorophenyl)-6-ethylpyrimidine], fluoropyrimethamine [2, 2,4-diamino-5-(4'-fluorophenyl)-6-ethylpyrimidine], fluoronitropyrimethamine [3, 2,4-diamino-5-(4'-fluoro-3'-nitrophenyl) -6-ethylpyrimidine], and methylbenzoprim [4, 2,4-diamino-5-[4'- (methylbenzylamino)-3'-nitrophenyl]-6-ethylpyrimidine]). The signals were identified mainly by correlating signals from bound and free ligands by using 2D exchange experiments. Analogues (such as 1 and 2) with symmetrically substituted phenyl rings give rise to 1H signals from four nonequivalent aromatic protons, clearly indicating the presence of hindered rotation about the pyrimidine-phenyl bond. Analogues containing asymmetrically substituted aromatic rings (such as 3 and 4) exist as mixtures of two rotational isomers (an enantiomeric pair) because of this hindered rotation and the NMR spectra revealed that both isomers (forms A and B) bind to the enzyme with comparable, though unequal, binding energies. In this case two complete sets of bound proton signals were observed. The phenyl ring protons in each of the two forms experience essentially the same protein environment (same shielding) as that experienced by the corresponding protons in bound pyrimethamine: this confirms that forms A and B correspond to two rotational isomers resulting from approximately 180 degrees rotation about the pyrimidine-phenyl bond, with the 2,4-diaminopyrimidine ring being bound similarly in both forms. The relative orientations of the two forms have been determined from NOE through-space connections between protons on the ligand and protein.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
92.
93.
Host modification of Sindbis virus sialic acid content influences alternative complement pathway activation and virus clearance 总被引:9,自引:0,他引:9
R L Hirsch D E Griffin J A Winkelstein 《Journal of immunology (Baltimore, Md. : 1950)》1981,127(5):1740-1743
Previous studies have shown that Sindbis virus, an enveloped alphavirus of the togavirus group, activates the alternative complement pathway in the absence of detectable antiviral immunoglobulin. The present studies examined the role of the host-determined sialic acid content of Sindbis virus on activation of the alternative complement pathway. Purified Sindbis virus grown in baby hamster kidney (BHK-SV) and in mosquito (MOSQ-SV) cells yielded virus with 10.2 and less than 2.0 nmol sialic acid/mg viral protein, respectively. Sindbis virus deficient in sialic acid (2.0 nmol sialic/mg) was also produced by treating the BHK-SV with neuraminidase (NANase-SV). When MOSQ-SV or NANase-SV was incubated in either C4DGPS or C2DHS, each consumed significantly more C3 than did BHK-SV, indicating that the ability of Sindbis virus to activate the alternative pathway is inversely related to its sialic acid content. Studies in vivo showed that virus deficient in sialic acid (MOSQ-SV) was cleared from the blood of mice much more efficiently than was virus rich in sialic acid (BHK-SV), after i.v. inoculation. Furthermore, when animals were depleted of C3 through C9 by cobra venom factor (CoVF) treatment, no differences in the clearance of high and low sialic acid-containing viruses were observed. Thus both the activation in vitro and complement-dependent clearance in vivo are significantly affected by the host-determined sialic acid content of Sindbis virus. 相似文献
94.
G. D. Griffin 《Journal of nematology》1980,12(4):287-293
Simultaneous inoculations of alfalfa with Meloidogyne hapla larvae and Ditylenchus dipsaci at 16, 20, 24, and 28 C did not depress penetration of either nematode in ''Nev Syn XX'' -a selection resistant to M. hapla and D. dipsaci, ''Vernal 298'' -a selection resistant to M. hapla and susceptible to D. dipsaci, ''Lahontan'' -a cultivar resistant to D. dipsaci and susceptible to M. hapla, and ''Ranger'' -a cultivar susceptible to both M. hapla and D, dipsaci. Infection with D. dipsaci depressed growth of susceptible ''Vernal 298'' and ''Ranger'' at all soil temperatures, except for ''Vernal 298'' at 16 C. Infection with M. hapla alone did not depress growth of any of the alfalfas. A combination of M. hapla and D. dipsaci resulted in a synergistic weight depression on ''Ranger'' at all soil temperatures. Inoculation of the four alfalfas with D. dipsaci 2, 4, 6, and 8 wk before inoculation with M. hapla at 16, 20, 24, and 28 C did not influence the resistance or susceptibility of ''Nev Syn XX,'' ''Lahontan,'' or ''Ranger.'' However, galling of ''Vernal 298'' by M. hapla was affected by soil temperature, plant age, and inoculation with D. dipsaci. 相似文献
95.
Identification and initial characterization of a new low-molecular-weight virus-encoded T antigen in a line of simian virus 40-transformed cells. 总被引:15,自引:11,他引:4 下载免费PDF全文
SV80 cells, a simian virus 40 (SV40)-transformed derivative of a strain of human fibroblasts, synthesize an 8-kilodalton anti-T reactive polypeptide in addition to large T and small t antigens. Although not observed during lytic infection carried out under a variety of conditions, an anti-T reactive molecule which comigrated with the SV80 8-kilodalton protein during sodium dodecyl sulfate-polyacrylamide gel electrophoresis was synthesized by one of five other SV40-transformed cell lines studied. The SV40 8-kilodalton protein was present in lysates of cells exposed to a brief pulse of radioactive methionine and did not accumulate during an extended chase period. This polypeptide could not by generated by mixing an unlabeled extract of SV80 cells with a labeled extract of infected monkey cells. The 8-kilodalton molecule reacts with antibody raised against homogeneous large T antigen, is present only in the cytoplasm, is not complexed with T, lacks DNA-binding properties, and is not phosphorylated. This protein could be translated in a cell-free system programmed by SV40-specific mRNA. At least two messenger species (approximately 19S and approximately 22S) directed its synthesis. Tryptic peptide analysis of [35S]methionine-labeled proteins demonstrated that the 8-kilodalton protein contains all eight of the common T/t peptides and one additional peptide not present in the maps of t or T. It lacks both of the t-unique peptides. The organization of the integrated viral sequences which encode this molecule was determined by restriction endonuclease analysis. In particular, SV80 cells contain at least two integrated SV40 genomes which are oriented in tandem, with an intervening cellular sequence.. 相似文献
96.
Martin A. Wasserman R.L. Griffin F.B. Marsalisi 《Prostaglandins & other lipid mediators》1980,20(4):703-715
Previously, we demonstrated that prostaglandin D2 (PGD2), a natural product of the endoperoxide PGH2, evoked bronchoconstriction when given I.V. to dogs (PROSTAGLANDINS :255–269, 1977). The present investigation in anesthetized dogs demonstrated that aerosols of PGD2 (0.001–0.1%) produced concentration-dependent increases in pulmonary resistance (RL) and decreases in dynamic lung compliance (CDYN) which were short-lived and equipotent to PGF2α. These alterations in pulmonary mechanics were partially, yet significantly, inhibited by atropine, thereby suggesting that at least a portion of the bronchoconstriction may be cholinergically mediated. Concomitant cardiovascular depressant effects of both PGD2 and PGF2α aerosols were much less and more variable than their bronchopulmonary effects.These results demonstrate a potent bronchoconstrictor effect of aerosolized PGD2 in dogs. PGD2 warrants further attention as a possible mediator of the bronchospasm seen in acute, reversible airways obstruction. 相似文献
97.
Olivia Hanson-Painton Martin J. Griffin Jordan Tang 《Biochemical and biophysical research communications》1981,101(4):1364-1371
Solutions of cytosolic proteins from rat liver contain benzo(a)pyrene solubilizing activity capable of serving as a carrier between solid state benzo(a)pyrene and microsomal cytochrome P450. Fractionation of benzo(a)pyrene-saturated cytosolic proteins on a Sephadex G-100 column or by sucrose density gradients produced benzo(a)pyrene peaks of about 46,000 daltons and a very high molecular weight material. The protein-bound benzo(a)pyrene obtained in both peaks was oxidized rapidly by microsomes in the presence of NADPH, indicating that the benzo(a)pyrene carrier activity is capable of presenting the substrate to the cytochrome P450. Liver cytosolic proteins from rats receiving intraperitoneal injection of [14C] benzo(a)pyrene was chromatographed on a column of Sephadex G-75. Radioactivity eluted at the same positions of the chromatogram as did the carrier activities described above. These results indicate that these benzo(a)pyrene carrier proteins may have an role in the metabolism of benzo(a)pyrene. 相似文献
98.
B W Griffin 《Archives of biochemistry and biophysics》1978,190(2):850-853
Horseradish peroxidase and metmyoglobin catalyze the H2O2-supported N-demethylation of N,N-dimethylaniline and N,N-dimethyl-p-toluidine. The catalytic activities of horseradish peroxidase are more than 100-fold larger than those of metmyoglobin or those previously reported for liver microsomal cytochrome P-450. Distinct free radical species of these N-methyl substrates were detected with both catalysts. These findings establish the general validity of a recently proposed free radical mechanism of oxidative N-demethylation (Griffin, B. W., and Ting, P. L., Biochemistry (1978), 2206–2211), which is quite different from that previously suggested for the analogous cytochrome P-450-dependent reactions. 相似文献
99.
Donald R. Griffin 《Animal behaviour》1976,24(2):421-427
Breeding choruses of Hyla crucifer and H. versicolor are loud enough to be audible to migrating birds up to at least 1 km from their source, both vertically and horizontally, provided that no large obstacles intervene. During May in south-eastern New York State sound pressure levels (A weighting) at altitudes of 200 to 965 m and slant ranges from the frogs of 225 to 1020 m varied from 28 to 52 dB SPL. 相似文献
100.
Colonization of rye green manure and peanut fruit debris by Aspergillus falvus and Aspergillus niger group in field soils. 总被引:1,自引:1,他引:0 下载免费PDF全文
Aspergillus flavus and Aspergillus niger group colonization of deep-plowed, decomposing rye green manure cover crops in peanut field soils was studied in four fields during 1972 and 1973; colonization of decomposing peanut fruits was studied in 1972 in two fields. A. flavus colonization of rye and peanut fruits was greater in soils of heavy texture, and an A. flavus population as high as 165 propagules per g of soil was observed in soil adjacent to rye, whereas A. flavus populations in soils not associated with rye were 18 propagules per g of soil or lower. Highest A. flavus populations in soil adjacent to decomposing peanut fruits were usually comparable to populations associated with rye. Little decomposing rye or peanut fruit colonization was generally observed by the A. flavus competitor, A. niger group. A. flavus may maintain or increase its inoculum potential by colonization of these and other moribund plant tissues. 相似文献