共查询到20条相似文献,搜索用时 281 毫秒
1.
K S Sundaram T A Venkitasubramanian 《Biochemical and biophysical research communications》1977,78(2):839-845
The effect of various sub-inhibitory concentrations of isoniazid on tryptophan uptake by H37Rv grown and was studied. Uptake, measured after 3 minutes of drug exposure was inhibited mildly by 0.1 μg/ml and 0.2 μg/ml concentration and completely by 0.3 μg/ml. However, with the minimal inhibitory concentration (MIC)7 of 0.5 μg/ml, not only inhibition but also a strong efflux of the preformed tryptophan pool were observed. The results are discussed in the light of the theory that isoniazid interferes with the cell wall mycolate synthesis. 相似文献
2.
17β-Hyd.roxyandrost-4-ene-3,11-dione was linked via its 3-(O-carboxymethyl) oxime to bovine serum albumin to give a conjugate which was used to generate antiserum in rabbits. The antiserum, at an overall dilution of 1 in 16,000, together with [1,2-3H] 17β-hydroxyandrost-4-ene-3,11-dione synthesized from [1,2-3H] cortisone have been used to develop a radioimmunoassay for the parent steroid. The assay incorporates a purification step in which serum or plasma extracts are chromatographed on silica gel layers bound to plastic or aluminium sheets and the steroid, containing zones cut out and eluted directly with assay buffer. The cross-reactivities of several steroids with the antiserum and the specificity, sensitivity, accuracy and precision of the assay are described. Blood sera from Immature male rainbow trout contain 0.2–0.4 μg/100 ml of 17β-hydroxyandrost-4-ene-3,11-dione. As male fish mature, serum levels rise sharply to reach values of 2 to >9 μg/100 ml. Levels in immature females rarely exceeded the assay sensitivity but serum from three ripe females showed low but detectable levels ( 0.2 μg/100 ml) of steroid. The assay has found application in sexing live fish for experimental purposes. 相似文献
3.
The mithramycin fluorescence procedure described by B. T. Hill and S. Whatley (1975, FEBS Lett., 56, 20–23) for DNA measurement tends to underestimate DNA concentrations in biological samples as compared to the results obtained by the diphenylamine reaction. This discrepancy disappears when DNA is first solubilized, by buffer containing heparin, from either cell homogenates or nuclear preparations. The optimal conditions for maximal fluorescence are 8 mm Mg2+, 10 μg/ml mithramycin, and heparin to DNA ratios ≥0.15 (). Background fluorescence is reduced 90% by dextran-coated charcoal adsorption of unbound mithramycin. The limit of sensitivity of the assay is 0.3 μg/ml and fluorescence is linear up to 30 μg DNA/ml. 相似文献
4.
G F Gerard C Gurgo D P Grandgenett M Green 《Biochemical and biophysical research communications》1973,53(1):194-201
Rifampicin and three rifamycin SV derivatives with different lipophilic side chains were tested as inhibitors of a number of purified enzymes including the α and αβ forms of RNA-directed DNA polymerase of avian myeloblastosis virus (AMV). (2,5-dimethyl-4-N-benzyl demethyl rifampicin), (O-n-octyloxime of 3-formyl rifamycin SV) and C-27 (rifamycin SV with a dicyclohexylalkyl substituted piperidyl ring at the 3-position) at concentrations less than 20 to 40 μg/ml completely inhibited the RNA- and DNA-directed DNA polymerase and RNase H activities of both AMV enzymes. Rifampicin was inactive at 100 μg/ml. When used against a variety of non-polymerizing enzymes such as alkaline phosphatase, glutamate-oxaloacetate transaminase, DNase I, and RNase A, these derivatives were inactive at drug concentrations between 100 and 200 μg/ml. Polynucleotide phosphorylase was inhibited slightly by all three derivatives. These results support the idea that rifamycin SV derivatives with appropriate 3-substituted side-chains are specific inhibitors of nucleic acid polymerizing enzymes. 相似文献
5.
Replication of the nuclear genome in yeast does not require concomitant protein synthesis 总被引:9,自引:0,他引:9
D H Williamson 《Biochemical and biophysical research communications》1973,52(3):731-740
Incorporation of 3H-adenine into nuclear DNA in a short pulse in mid-S in a synchronised culture of was unaffected by the presence of 100 μg/ml cycloheximide. However, colorimetric DNA analyses showed that entry into S was completely blocked by adding the drug at times earlier than about 10 min before initiation of replication. Cell autoradiography of cultures labelled in various regimes showed that at this time there is a cycloheximide-transition point at which the cell acquires the capacity to both initiate and complete a whole round of replication in the presence of 100 μg/ml of cycloheximide. Thus, all the proteins required for passage through one S period are made in advance of initiation. 相似文献
6.
M.S. Kang James P. Spencer Alan D. Elbein 《Biochemical and biophysical research communications》1978,82(2):568-574
Amphomycin inhibits the incorporation of mannose from GDP-[14C]mannose and GlcNac from UDP-[3H]GlcNAc into lipid-linked saccharides by either a particulate or a solubilized enzyme fraction from pig aorta. The solubilized enzyme was much more sensitive to the antibiotic than was the particulate fraction with 50% inhibition being observed at 8–15 μg of amphomycin. Although the antibiotic inhibited mannose transfer from GDP-[14C]mannose into mannosyl-phosphoryl-dolichol, lipid-linked oligosaccharides and glycoprotein, the synthesis of mannosyl-phosphoryl-dolichol was much more sensitive to amphomycin. Amphomycin also inhibited the incorporation of mannose from GDP-[14C]mannose into mannosyl-phosphoryldecaprenol in particulate extracts of . 相似文献
7.
J D Kemp 《Biochemical and biophysical research communications》1976,69(3):816-822
Octopine [N2-(D-1-carboxyethyl)-L-arginine] was detected in all tobacco and sunflower crown gall tumors incited by (E. F. Sm. and Town.) Conn strain B6 at levels between 1 and 2.5 μmoles/20 g fresh weight. Most tissue cultures derived from plant tumors contained octopine at levels between 0.3 and 1 μmole/20 g fresh wt. Normal plant tissues and tissue cultures derived from normal tissues contained no detectable octopine when assayed by a [3H] arginine incorporation technique designed to detect low levels of octopine (less than 0.5 nmole/20 g fresh wt). 相似文献
8.
The transformation efficiency of prepared by the calcium-heat shock procedure has been increased sixfold by growth of cells in 0.5 M sucrose and the addition of 1 μg/ml lysozyme along with DNA. A mutant of , has been selected which has a tenfold increased efficiency of transformation. 相似文献
9.
The effects of endometrium on metabolism of [3H]-arachidonic acid ([3H]-AA) by bovine blastocysts recovered on day 19 postmating were studied . Blastocysts (n = 12) and endometrial slices were assigned to four incubation groups. In group 1, blastocysts were incubated alone; group 2, endometrial slices were incubated alone; group 3, blastocysts were incubated with endometrial slices; group 4, blastocysts were incubated in 7.5 ml fresh incubation medium plus 7.5 ml frozen-thawed medium from endometrial incubations. In all groups, tissues were incubated in 15 ml modified minimum essential medium (MEM) containing 5 μCi of [3H]-AA and 200 μg radioinert arachidonic acid for 24 h at 37°C in an atmosphere of 50% N2:45% O2:5% CO2. For incubation controls, 5 μCi of [3H]-AA were added to 15 ml MEM and incubated at the same time as tissues from each cow. To evaluate metabolism of [3H]-AA, [3H]-AA and its metabolites were extracted from aliquots of MEM and separated on columns of Sephadex LH-20. Most (78.3 ± 3.2%) of the radioactivity (dpm) in the incubation controls was recovered as [3H]-AA, indicating that there was little breakdown of [3H]-AA in the absence of tissue. Blastocysts produced compounds that migrated with [3H]-13,14-dihydro-15-keto-PGF2α ([2H]-PGFM), [3H]-PGE2 and [3H]-PGF2α. Endometrial slices metabolized very little of the [3H]-AA. Data from groups 1 and 4 were combined (group ) for analysis because the distribution of dpm did not differ between the two groups. In group 3, blastocysts and endometrial slices incubated together tended(P<.10) to produced more [3H]-PGE2 than did group , there tended to be less (P<.10)_[3H]-PGF2α, and there was more (P<.05) [3H]-PGFM than in group . Neither endometrial secretions nor endometrial slices altered the proportion of [3H]-AA metabolized by blastocysts. Endometrial slices appear capable of metabolizing [3H]-PGF2α synthesized by blastocysts, and capable of directing blastocyst metabolism of [3H]-AA away from synthesis of [3H]-PGF2α and toward synthesis of [3H]-PGE2. It is postulated that the endometrium has an important role in regulating the amounts and ratios of prostaglandins in th uterine lumen during early prenancy in cows. 相似文献
10.
A. Frisch H. Levkowitz A. Loyter 《Biochemical and biophysical research communications》1976,72(1):138-145
Tunicamycin, a glucosamine-containing antibiotic inhibited the conjugation process of Tetrahymena pyriformis. Sexual pairing was prevented completely when 1.5 μg/ml of tunicamycin was added to a mixture of the two mating types. Tunicamycin caused preferential inhibition of glycoprotein synthesis in Tetrahymena pyriformis. At 1.5 μg/ml and 6 μg/ml tunicamycin inhibited by 40% and 60% respectively [3H]-glucosamine incorporation into material precipitated by ethanol, while it did not affect [14C]-leucine incorporation. Cell division was also inhibited when the drug was added either to the regular growth medium or to the starvation medium. 相似文献
11.
The number of silk fibroin genes per genome in the silkworm Bombyx mori has been determined by hybridization using fibroin [125I]mRNA. The purified [125I]mRNA had an oligonucleotide pattern after RNAase T1 digestion which was characteristic of fibroin mRNA (Suzuki &; Brown, 1972) and it hybridized specifically to DNA with a G + C content expected for a fibroin gene. Thermal denaturations indicated that these hybrids were mismatched by about 3%, which probably indicates some variation among the sequences encoding the internal repetitions of the fibroin protein.The concentration of fibroin gene sequences in B. mori DNA was measured by saturation hybridization of [125I]mRNA to filter bound DNA. The same saturation level of 1.8 × 10?5 μg mRNA per μg DNA was calculated from data obtained with unfractionated DNA and with fibroin gene sequences which had been separated from bulk B. mori DNA by actinomycin centrifugation. Scatchard plots of the subsaturation data extrapolated to an identical saturation value. Internal reiteration of the fibroin mRNA molecule was apparent from the high association constant of hybridization. An exhaustive hybridization experiment showed that such repetitions comprise at least 90% of each mRNA molecule. The saturation value, in conjunction with the genome DNA content and the mRNA size, indicated the presence of only one fibroin gene per haploid B. mori genome.Hybridization of actinomycin fractionated DNA indicated that fibroin mRNA can form hybrids with DNA that bands with bulk B. mori DNA. These hybrids appear to involve DNA which is related to, but distinguishable from, true fibroin gene sequences. The fibroin gene-related sequences form mismatched hybrids with the mRNA, are much shorter than the fibroin gene and are dispersed in B. mori DNA of much lower G + C content, and there are many copies of these sequences per B. mori genome. 相似文献
12.
Ward R. Rice Jeffrey A. Whitsett 《Biochimica et Biophysica Acta (BBA)/Molecular Cell Research》1984,805(3):261-267
Release of [3H]phosphatidylcholine from pulmonary Type II epithelial cells was stimulated by terbutaline, forskolin and cytochalasin D. Compound inhibited both basal and agonist-stimulated release of [3H]PC. The IC50 for inhibition by compound was 1–2 μg/ml, and was similar for inhibition of both basal and stimulated release of [3H]phosphatidylcholine. Inhibitory effects of were noted following a 1 h exposure to compound and persisted up to 3 h. The inhibitory effect of compound was entirely reversed by removing compound from the external milieu. Compound had no effect on cytosolic cyclic AMP levels or lactate dehydrogenase release. Inhibition of surfactant release produced by compound was unaffected by changes in extracellular calcium concentrations. Compound is a non-toxic inhibitor of phosphatidylcholine release from Type II epithelial cells. 相似文献
13.
It is demonstrated by direct measurement of surface radioactivity that the cationic polypeptide antibiotic polymyxin B is specifically adsorbed to negatively charged lipid monolayers. The latter attracted the following amounts of the biologically active derivative (PX): lipid A from Proteus mirabilis, 0.17; phosphatidic acid, 0.12; phosphatidylglycerol and phosphatidylserine, 0.11; dicetylphosphate, 0.107; sulfoquinovosyldiglyceride, 0.104; phosphatidylinositol and cardiolipin, 0.095; and phosphatidylethanolamine, 0.017 μg/cm2. Adsorption of PX to phosphatidylcholine, monogalactosyldiglyceride and stearylamine was almost or completely zero. Total lipids from Escherichia coli adsorbed 0.057 in comparison to 0.051 μg PX/cm2 of an artificial mixture of phosphatidylethanolamine/phosphatidylglycerol/cardiolipin in the proportions 75 : 25 : 5. The concentration of the surface active PX at the air/water interphase was 0.091 μg/cm2. These saturation surface concentrations of PXat lipid monolayers were reached at 1 μg/ml bulk concentrations in 2 mM NaCl/1 mM Tris · HCl, pH 7.2. They decreased with decreasing surface charge density of the adsorbing monolayer. In an experiment with cardiolipin/phosphatidylethanolamine mixtures it was shown that two molecules of cardiolipin induced adsorption of one molecule PX giving a 1 : 1 ratio with regard to positive and negative charges. This could be due to a similar charge density of about one charge per 40–50 Å2 in PX and lipid bilayers composed of phospholipids. The electrostatic PX-lipid interaction was severely inhibited by 10?2 and 10?1 M Ca2+ and Na+, respectively. It is discussed that the specificity of PX against Gram-negative bacteria is caused by the occurrence of lipid A, phosphatidylglycerol and cardiolipin at the cell surface of these microorganisms. 相似文献
14.
M.E. Eldefrawi N. Shaker N.A. Mansour J.E. Warnick E.X. Albuquerque 《Life sciences》1981,29(10):1033-1037
Biochemical and electrophysiological studies were conducted on the electric organ of the electric fish of the Nile, , in order to determine if transmission was chemically mediated. There was no binding of [3H] acetylcholine, [3H] quinuclidinyl benzilate or [3H]-perhydrohistrionicotoxin; but low acetylcholinesterase activity was observed, as was binding of [125I] α-bungarotoxin. The latter binding was detectable at 0.85 ± 0.07 pmol/g tissue, and was totally inhibited by 1 μM α-bungarotoxin or 100 μM d-tubocurarine. A tetrodotoxin-sensitive action potential was measured which was Na+- dependent. Depolarization (30–40 mV) was caused by carbamylcholine, and this was blocked by d-tubocurarine or α-bungarotoxin. The data suggest that this electric organ which may be a rich source for electrically excitable channels, is innervated by nicotonic cholinergic motoneurons, but the concentrations of acetylcholine receptors and acetylcholinesterase are very low. 相似文献
15.
A L Flowers D A Hessinger 《Biochemical and biophysical research communications》1981,103(3):1083-1091
Nematocyst venom from Portuguese Man-of War ( sp.) tentacles causes isolated rat peritoneal mast cells to release histamine. Extent of histamine release is dose-dependent (K0.5 = 6.1 μg venom/ml) and attains 100% at high doses of venom. Release is independent of intra- and extracellular calcium levels and does not depend upon a cellular supply of ATP. The rate of histamine release is temperature-dependent and the extent of release is maximized broadly over the range of 10–30°C. The cytoplasmic marker lactate dehydrogenase, is released concomitantly with histamine but is more sensitive to the venom (K0.5 = 2.1 μg/ml). Antimycin A, while it does not significantly affect venom-induced histamine release, increases the sensitivity of lactate dehydrogenase release (K0.5 = 0.2 μg/ml). We conclude that nematocyst venom induces the release of histamine from mast cells by a cytolytic mechanism and that this action is antagonized by an intracellular, energy-requiring process. 相似文献
16.
Brinolase, a thrombolytic fungal protease capable of forming vasoactive kinins, has been shown to hydrolyze kinins after their formation. Using synthetic bradykinin as a substrate, the kinetics and mechanism of hydrolysis have been elucidated, evidently explaining the apparently low kinin formation , Bradykinin hydrolysis proceeded rapidly with a pH optimum of 7.0–7.5, and a half-life of 5.1 min, using 250 ng/ml bradykinin and 50 μg/ml brinolase. The Km was 3.2×10?6 M and the Vmax was 4.6 × 10?8 mol/liter/min, using 5 μg/ml brinolase. Two-dimensional paper fractionation of the brinolase-bradykinin digest revealed the presence of free arginine amongst the five peptide fragment spots. 相似文献
17.
(1) In order to assess the possible role of 3′,5′-(cyclic)adenosine monophosphate (cAMP) in the control of glucose transport, the effect of the nucleotide or agents known to increase its intracellular concentration on sugar transport or 45Ca2+ washout were characterized in epididymal fat pads, free fat cells and soleus muscles of the rat. (2) When added to the incubation medium, cAMP (0.1–2.0 mM) stimulated washout from fat pads. This effect was abolished by cytochalasin B, and additive to that induced by submaximal (10–25 μU/ml), but not by supramaximal (10 mU/ml) concentrations of insulin. (3) cAMP (2 mM) stimulated the conversion of [U-14C]glucose into CO2 and triacylglycerols. This effect was additive to that of insulin (100 μU/ml). (4) ACTH, glucagon, adrenaline, noradrenaline and salbutamol, which are all known to increase the cAMP content of adipose tissue, stimulated the washout of and 45Ca2+ from preloaded fat pads. The fractional losses of the two isotopes were significantly correlated (, ). (5) In free fat cells, adrenaline (10?6 M) and salbutamol (10?5 M) stimulated the uptake of , and salbutamol (10?5 M) did not interfere with the stimulating effect of insulin (25 μU/ml) on sugar uptake. (6) In rat soleus muscles, adrenaline and salbutamol produced a dose-dependent stimulation of the washout of and 45Ca2+. The effect of adrenaline on sugar efflux was abolished by propranolol. (7) It is concluded that the activation of the glucose transport system by insulin is unlikely to be mediated by a drop in the cellular concentration of cAMP. An increase in cAMP brought about by β-adrenoceptor agonists or lipolytic hormones may induce a mobilization of calcium ions from cellular pools into the cytoplasm, which in turn leads to the activation of the glucose transport system demonstrated in the present as well as in several earlier studies. 相似文献
18.
B L Currie J Warberg K Folkers 《Biochemical and biophysical research communications》1977,79(4):1207-1211
The hypocholesterolemic drug clofibrate (ethyl-α--chlorophenoxyisobutyrate) was found to strongly suppress 3-hydroxy-3-methylglutaryl coenzyme A reductase (EC 1.1.1.34) activity in cultured mouse L cells at concentrations of 20 – 50 μg/ml. The half-life () of the reductase (approximately 120 min) was strongly reduced when L cells were incubated with cycloheximide plus a maximal inhibitory concentration of clofibrate (50 μg/ml), resulting in a value of 10 min. Preliminary kinetic analysis of the inhibition suggested that clofibrate increased the rate of inactivation (or degradation) of the reductase without affecting the rate of enzyme synthesis. 相似文献
19.
Cerulenin resistance in a cerulenin-producing fungus. Isolation of cerulenin insensitive fatty acid synthetase 总被引:3,自引:0,他引:3
A Kawaguchi H Tomoda S Okuda J Awaya S Omura 《Archives of biochemistry and biophysics》1979,197(1):30-35
Cerulenin, an antifungal antibiotic isolated from a culture filtrate of Cephalosporium caerulens, is a potent inhibitor of fatty acid synthetase systems. This antibiotic specifically blocks the activity of β-ketoacyl thioester synthetase (condensing enzyme). The mechanism of the resistance of C. caerulens to cerulenin was investigated. The rate of growth in medium containing up to 100 gmg/ml cerulenin was as rapid as that in cerulenin-free medium. At a cerulenin concentration of 300 μg/ml, the rate of growth was still more than half that of the control. The addition of cerulenin (200 μg/ml) to a culture of growing cells has almost no effect on the incorporation of []acetate into cellular lipids. Fatty acid synthetase was purified from C. caerulens to homogeneity. Properties of this fatty acid synthetase were almost the same as those of yeast fatty acid synthetase except for the sensitivity to cerulenin. C. caerulens synthetase is much less sensitive to cerulenin than fatty acid synthetases from other sources. These findings suggested that the insensitivity of C. caerulens fatty acid synthetase plays an important role in the cerulenin resistance of this fungus. 相似文献
20.
Klaus Gietzen Peter Adamczyk-Engelmann Andreas Wüthrich Anka Konstantinova Hermann Bader 《生物化学与生物物理学报:生物膜》1983,736(1):109-118
Compound , a condensation product of N-methyl-p-methoxyphenethylamine with formaldehyde, is composed of a family of cationic amphiphiles differing in the degree of polymerization. Compound was found to be a potent inhibitor of the calmodulin-activated fraction of brain phosphodiesterase and red blood cell Ca2+-transport ATPase, with IC50 values of 0.3 and 0.85 μg/ml, respectively. However, the basal activity of both enzymes is not at all suppressed by the drug at concentrations up to 300 μg/ml. Inhibition of Ca2+ transport into inside-out red blood cell vesicles by compound follows a similar pattern in that basal, calmodulin-independent, transport is also not affected by the drug. Kinetic analysis revealed that the stimulation of Ca2+-transport ATPase induced by calmodulin is inhibited by compound according to a competitive mechanism. It was demonstrated that the inhibitory constituents of compound bind to calmodulin in a Ca2+-dependent fashion. Comparison of the specificity of several anti-calmodulin drugs showed that compound is the most specific inhibitor of the calmodulin-dependent fraction of red blood cell Ca2+-transport ATPase that has been described hitherto. In addition, compound was found to be a rather specific inhibitor of the calmodulin-induced activation of Ca2+-transport ATPase when compared with the stimulation induced by an anionic amphiphile or by limited proteolysis. Half-maximal inhibition of the activity stimulated by oleic acid or mild tryptic digestion required 8- and 32-times higher concentrations of compound , respectively, compared with the calmodulin-dependent fraction of the ATPase activity. Moreover, calmodulin-independent systems as rabbit skeletal muscle sarcoplasmic reticulum Ca2+-transport ATPase or calf cardiac sarcolemma (Na+ + K+)-transport ATPase are far less influenced by compound as compared with trifluoperazine and calmidazolium. Because of its high specificity compound is proposed to be a promising tool for studying calmodulin-dependent processes. 相似文献