共查询到20条相似文献,搜索用时 15 毫秒
1.
W. Wackernagel 《Biochemical and biophysical research communications》1973,51(2):306-311
Genetic transformation of for various chromosomal markers was accomplished by (i) using recipient cells that lack the DNase but were recombination proficient due to or mutations and (ii) treating the recipient cells with CaCl2 at a concentration that facilitates transfection by λ DNA. Cotransformation of three markers () was found to depend on the molecular weight of the transforming DNA. 相似文献
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3.
The technique of laser Doppler electrophoresis was applied for the study of the surface charge properties of (Na+,+)-ATPase containing microsomal vesicles derived from guinea-pig kidney. The influence of pH, the screening and binding of uni- and divalent cations and the binding of ATP show: (1) one net negative charge per protein unit with a ; (2) deviation from the Debye relation between surface potential and ionic strength for univalent cations, with no difference in the effect of Na+ and K+; (3) Mg2+ binds with an association constant of while ATP binds with an apparent for 1 mM Nacl, 0.2 mM KCI, 0.1 mM MgCl2, 0.1 mM Tris-HCI (pH 7.3). The binding is weaker at higher Mg2+ concentrations. There is no ATP binding in the absence of Mg2+. In addition, the average vesicle size derived from the linewidth of the quasi-elastic light scattering spectrum is . In the presence of ATP a reduction in size is observed. 相似文献
4.
Peter Nicholls 《BBA》1976,430(1):13-29
1. Formate inhibits cytochrome oxidase activity both in intact mitochondria and submitochondrial particles, and in isolated cytochrome . The inhibition increases with decreasing pH, indicating that HCOOH may be the inhibitory species.2. Formate induces a blue shift in the absorption spectrum of oxidized cytochrome ) and in the half-reduced species (). Comparison with cyanide-induced spectral shifts, towards the red, indicates that formate and cyanide have opposite effects on the spectrum, both in the fully oxidized and the half-reduced states. The formate spectra provide a new method of obtaining the difference spectrum of minus , free of the difficulties with cyanide (which induces marked high → low spin spectral shifts in cytochrome ) and azide (which induces peak shifts of cytochrome towards the blue in both α- and Soret regions).3. The rate of formate dissociation from cytochrome is faster than its rate of dissociation from , especially in the presence of cytochrome . The for formate inhibition of respiration is a function of the reduction state of the system, varying from 30 mM (100% reduction) to 1 mM (100% oxidation) at pH 7.4, 30 °C.4. Succinate-cytochrome reductase activity is also inhibited by formate, in a reaction competitive with succinate and dependent on [formate]2.5. Formate inhibition of ascorbate plus oxidation by intact rat liver mitochondria is partially released by uncoupler addition. Formate is permeable through the inner mitochondrial membrane and no differences in ‘on’ or ‘off’ inhibition rates were observed when intact mitochondria were compared with submitochondrial particles.6. NADH-cytochrome reductase activity is unaffected by formate in submitochondrial particles, but mitochondrial oxidation of glutamate plus malate is subject both to terminal inhibition at the cytochrome level and to a slow extra inhibition by formate following uncoupler addition, indicating a third site of formate action in the intact mitochondrion. 相似文献
5.
Yuji Kamiya Akira Sakurai Nobutaka Takahashi 《Biochemical and biophysical research communications》1980,94(3):855-860
Rhodotorucine which induces mating tube formation of cells in is metabolized rapidly by cells. By use of labeled rhodotorucine , the degradation was found to be proteolytic. Two peptide fragments Tyr-Pro-Glu-Ile-Ser-Trp-Thr-Arg and Asn-Gly-Cys(S-farnesyl) were identified as the metabolites. Proteolysis of the pheromone mainly occurred on the cell surface. Culture filtrate of cells at log phase did not metabolize rhodotorucine . 相似文献
6.
Short, mild treatments of sarcoplasmic reticulum vesicles with aqueous from methanol to caused an inhibition of calcium uptake and an enhancement of ATPase activity. The treatments increased both calcium-dependent (extra) ATPase activity and calcium-independent (basic) ATPase activity of vesicles. The apparent initial reaction rate of ATPase of vesicles was about twice that of control vesicles. With increasing number () of carbon atoms of the , the maximum increment of ATPase activity increased, and both the alcohol concentration () required to inhibit calcium uptake by 50% and the alcohol concentration () required to enhance ATPase activity by 50% of the maximum increment of ATPase activity decreased as follows. The ratio, to , was constant for all values. The apparent free energy of binding of the methylene groups of to sarcoplasmic reticulum vesicles was evaluated (?796 cal/mole) and compared with data from the partition of in octanol and water (?670 cal/mole). The effects of on membrane vesicles are discussed on the basis of these data. 相似文献
7.
Kathryn L. Desphande Jon R. Katze James F. Kane 《Biochemical and biophysical research communications》1980,95(1):55-60
Glutamate synthase, an important enzyme in the assimilation of ammonia, was measured in cultures of grown with different nitrogen sources. An attempt was made to correlate the specific activity to the intracellular levels of five metabolites of glutamate metabolism: aspartate, glutamate, glutamine, alanine and . An inverse relationship was found between the activity of glutamate synthase and the pool level of glutamine. We propose that the intracellular concentration of glutamine is an important element in controlling the level of glutamate synthase. 相似文献
8.
Yukihiro Shoyama Yasuo Kishimoto 《Biochemical and biophysical research communications》1976,70(4):1035-1041
A doubly labeled 3-ketoceramide, [1-14C] lignoceroyl [1-3H2] 3-ketosphingosine ( ratio, 3.61) was injected into the left ventricle of rat heart. The ceramide isolated from the livers of the animals after 1 hr incubation contained an equal ratio of 3.60. This finding strongly supports the existence for direct conversion of 3-ketoceramide to ceramide in rat liver. 相似文献
9.
Satoshi Ōmura Hiroie Ohno Tsuneo Saheki Masakazu Yoshida Akira Nakagawa 《Biochemical and biophysical research communications》1978,83(2):704-709
Elasnin, a new human granulocyte elastase inhibitor, has been isolated from KM-2753. Elasnin is a neutral, lipophilic colorless and viscous oil (, , λmaxEtOH 291 nm (ε, 7760)). The molecular formula was C24H40O4 (M.W.: 392) as determined by its elemental analysis and mass spectrum. Elasnin inhibits markedly human granulocyte elastase, but is almost ineffective for pancreatic elastase, trypsin, chymotrypsin, thermolysin and papain. 相似文献
10.
Roxanne Deslauriers Harold C. Jarrell R.Andrew Byrd Ian C.P. Smith 《Biochemical and biophysical research communications》1980,95(3):1211-1217
studies of have shown that encysting cells release polyphosphate into the encystment medium. Mature cysts contain low levels of polyphosphate, as do vegetative cells. Young cysts (7 days) show detectable levels of nucleotide diphosphates and triphosphate similar to those observed in vegetative cells. Mature cysts (90 days) show only excreted polyphosphate as well as a component which has a chemical shift of a phosphodiester. The inorganic phosphate peak in the cyst shows that the cyst milieu is liquid-like and that the intracellular environment maintains a pH between 6 and 7.5 in the presence of extracellular values from 4 to 9. 相似文献
11.
Maura Floreani Anna Chiara Bonetti Francesca Carpenedo 《Biochemical and biophysical research communications》1981,101(4):1337-1344
Intact synaptosomes prepared from rat brain were incubated with phosphatidylserine vesicles. The synaptosomes incorporated the phospholipid in proportion to its concentration in the preincubation medium. The activity of membrane-bound enzyme ATPase increased proportionally after treatment with phosphatidylserine liposomes.When breaking phosphatidylserine-enriched synaptosomes by osmotic shock or by sonication and when preparing synaptosomal membranes, the expected increase of ATPase activity was not seen. Therefore, cellular integrity was fundamental in order to see the effect of phosphatidylserine on ATPase activity. 相似文献
12.
J.J. Schrijen A. Omachi W.A.H.M. Van Groningen-Luyben J.J.H.H.M. De Pont S.L. Bonting 《生物化学与生物物理学报:生物膜》1981,649(1):1-12
(1) The total phospholipid content of a gradient purified ()-ATPase preparation from pig gastric mucosa is 105 μmol per 100 mg protein, and consists of 29% sphingomyelin, 29% phosphatidylcholine, 28% phosphatidylethanolamine, 10% phosphatidylserine and 4% phosphatidylinositol. The cholesterol content corresponds to 50 μmol per 100 mg protein. (2) Treatment with phospholipase C (from Clostridium welchii and Bacillus cereus) results in an immediate decrease of the phosphate content. Up to 50% of the phospholipids are hydrolyzed by each phospholipase C preparation alone, without further hydrolysis by increased phospholipase concentration or prolonged incubation time. Combined treatment with the two phospholipase C preparations, sequentially or simultaneously, hydrolyzes up to 65% of the phospholipids. (3) The ()-ATPase and K+ stimulated activities are decreased proportionally with the total phospholipid content, indicating that these enzyme activities are dependent on phospholipids. (4) Phospholipase C treatment does not change optimal pH, value for ATP and temperature dependence of the gastric ()-ATPase, but slightly decreases the value for K+. (5) Phospholipase C treatment lowers the binding and phosphorylation capacities, suggesting that inactivation occurs primarily on the substrate binding level. (6) Most of the results can be understood by assuming that hydrolysis of the phospholipids by phospholipase C leads to aggregation of the membrane protein molecules and complete inactivation of the aggregated ATPase molecules. 相似文献
13.
The specific synthesis of F mRNA directed by the F gene carried on the specialized transducing bacteriophage F, performed , is described with the use of an S180 extract from a strain carrying R?. Synthesis of F mRNA is biphasic at approximately 7 minutes. The regulation of F mRNA synthesis by the specific arginine holorepressor present in an S180 extract prepared from a strain carrying the R+ allele is described. 相似文献
14.
This investigation was principally undertaken to test the ionic gradient hypothesis as applied to active uptake in the rabbit kidney cortical slice preparation. Efflux of from the slice was shown to be independent of external Na+ concentration. Transferring slices from a low sodium preincubation to a high sodium incubation medium containing increased intracellular concentrations of both Na+ and K+, and accumulation occurred. Transferring slices from a low sodium preincubation to a high sodium incubation medium containing ouabain and resulted in a net increase in intracellular Na+ concentration but no accumulation occurred. Different combinations of preincubation and incubation media gave a high to low array of intracellular Na+ concentrations and these directly reflected their respective uptake. These results suggest that the Na+-gradient hypothesis does not adequately explain the transport of organic acids in rabbit kidney. These results also suggest that Na+ possibly has an intracellular role through its stimulation of channeled to energizing the accumulative mechanism. 相似文献
15.
A functionally active ubiquinone derivative has been synthesized for the identification of the ubiquinone binding protein in ubiquinol-cytochrome reductase. After photolysis, the 14C activity was found to be specifically associated to proteins with mobilities relative to cytochrome of 0.841 and 0.475 in the sodium dodecylsulfate polyacrylamide gel electrophoresis of the Weber and Osborn system. These two proteins have previously been identified as cytochromes. The 14C activity distribution pattern was observed to be identical in the presence or absence of phospholipids during the photolysis. Antimycin A also produces no change in the 14C activity distribution among the proteins of this enzyme complex. 相似文献
16.
Thermotropic properties of purified cytochrome and cytochrome have been studied by differential scanning calorimetry under various conditions. Both cytochromes exhibit a single endothermodenaturation peak in the differential scanning calorimetric thermogram. Thermodenaturation temperatures are ionic strength, pH, and redox state dependent. The ferrocytochromes are more stable toward thermodenaturation than the ferricytochromes. The enthalpy changes of thermodenaturation of ferro- and ferricytochrome are markedly dependent on the ionic strength of the solution. The effect of the ionic strength of solution on the enthalpy change of thermodenaturation of cytochrome is rather insignificant. The formation of a complex between cytochromes and at lower ionic strength causes a significant destabilization of the former and a slight stabilization of the latter. The destabilization of cytochrome upon mixing with cytochrome was also observed at high ionic strength, under which conditions no stable complex was detected by physical separation. This suggests formation of a transient complex between these two cytochromes. When cytochrome was complexed with phospholipids, no change in the thermodenaturation temperature was observed, but a great increase in the enthalpy change of thermodenaturation resulted. 相似文献
17.
P.N. Bandyopadhyay Maharani Chakravorty 《Biochemical and biophysical research communications》1976,71(2):644-650
Infection of with the mutant of bacteriophage P22 leads to a rapid and severe efflux of intracellular leucine. The superinfection exclusion () genes of P22 interfere with the function of gene, the product(s) of which is speculated to be an internal protein of phage P22. 相似文献
18.
《Biochimica et Biophysica Acta (BBA)/General Subjects》1983,757(3):342-351
The existence of metabolically distinct pools of in Saccharomyces cerevisiae and isolated rat hepatocytes was investigated. Utilizing a relatively long labeling period with [methyl-14C]methionine, a metabolically ‘stable’ pool was labeled. A subsequent short labeling with [methyl-3H]methionine selectively labeled a putative metabolically ‘labile’ pool. The existence of these distinguishable pools was ascertained by following the 3H and 14C label disappearance in during the chase-period in label-free media containing cycloleycine to prevent futher synthesis of . In both yeast and hepatocytes, the ratio in decreased sharply. The individual 3H and 14C decrease in showed values of 3 and 8 min for yeast and 4 and 18 min for hepatocytes. The results strongly indicate that at least two metabolically distinct pools actually do exist in both systems. Subcellular fractionation revealed that the ‘labile’ pool exist in the cytosol for both yeast and hepatocytes while the ‘stable’ pool exists in the vacuolar and the mitochondrial fraction for the yeast and hepatocytes respectively. The pools were also studied in normal yeast under anaerobic chase condition and petite mutant yeast. Sharply contrasting with aerobically chased normal yeast, both showed closely parallel 3H and 14C decreases in . 相似文献
19.
Chana Vinkler 《Biochemical and biophysical research communications》1981,99(4):1095-1100
The of ADP for photophosphorylation in lettuce chloroplasts was measured both at various light intensities and in the presence of various uncoupler (nigericin + K+) concentrations. Lowering the light intensity results in both, a decrease in the rate of phosphorylation and a several fold in the of ADP for the reaction. However, when increasing concentrations of the uncoupler nigericin + K+ are employed, the rate of photophosphorylation is decreased but a several-fold in the of ADP for the reaction is observed. The results are discussed in terms of the chemiosmotic hypothesis. It is suggested that these effects might indicate the existence of a mechanism controlling the rate of ATP formation which is different than the formation of the electrochemical gradient. 相似文献
20.
Anthony E. Pegg 《Biochemical and biophysical research communications》1978,84(1):166-173
Extracts from various rat tissues were incubated with [3H]methylated DNA or chromatin in order to compare their abilities to catalyze the removal of labeled -methylguanine from acid precipitable DNA. Liver extracts had the greatest activity. Kidney extracts had about 35% of the activity in liver and extracts from lung, colon, small intestine and brain were much less active. The enzyme responsible for this reaction does not appear to be an -glycosidase because no labeled -methylguanine could be detected in the supernatant fraction even though more than 50% of this base was lost from the DNA. The released radioactivity was present as methanol which is consistent with the possibility that the reaction may involve a demethylase action on either the DNA substrate or an oligonucleotide derived from it. 相似文献