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121.
When grown with nitrate as terminal electron acceptor both the soluble (periplasm, cytoplasm) and the membrane fraction of Spirillum strain 5175 exhibited high nitrite reductase activity. The nitrite reductase obtained from the soluble fraction was purified 76-fold to electrophoretical homogeneity. The enzyme reduced nitrite to ammonia with a specific activity of 723 mol NO
inf2
sup-
× (mg protein × min)-1. The molecular mass was 58±1 kDa by SDS-PAGE compared to 59±2 kDa determined by size exclusion chromatography under nondenaturing conditions. The enzyme (as isolated) contained 5.97±0.15 heme c molecules/Mr 58 kDa. The absorption spectrum was typical for c-type cytochrome with maxima at 280, 408, 532 and 610 nm (oxidized) and at 420, 523 and 553 nm (dithionite-reduced). The enzyme (as isolated) exhibited a complex set of high-spin and lowspin ferric heme resonances with g-values at 9.82, 3,85, 3.31, 2.95, 2.30 and 1.49 in agreement with data reported for electron paramagnetic resonance spectra of nitrite reductases from Desulfovibrio desulfuricans, Wolinella succinogenes and Escherichia coli.Abbreviations DNRA
dissimilatory nitrate reduction to ammonia
- EPR
electron paramagnetic resonance
- PAGE
polyacrylamide gel electrophoresis
- NaPi
sodium phosphate
- SDS
sodium dodecylsulfate 相似文献
122.
Daniel Scherman 《Journal of neurochemistry》1986,47(2):331-339
The objective of the present study was to estimate extracellular pH (pHe) and intracellular pH (pHi) during near-complete forebrain ischemia in the rat, and to evaluate the relative importance of lactic acidosis and rise in tissue Pco2 (Ptco2) in causing pHe and pHi to fall. The animals, which were ventilated, normoxic, normocapnic, and normothermic, were subjected to 15 min of ischemia, either without or with 30-60 min of recirculation. Ptco2 was measured with a tissue electrode, pHe with a double-barrel liquid ion-exchanger microelectrode, changes in extracellular fluid (ECF) volume by impedance measurements, tissue CO2 content by a microdiffusion technique, and labile tissue metabolites by enzymatic fluorometric methods. Ischemia caused Ptco2 to rise to between 95 and 190 mm Hg (mean 149 mm Hg), and pHe to fall by 0.45-1.05 units (mean 0.70 units). During recovery, Ptco2 normalized within 5 min and pHe after 15-30 min. During ischemia, high-energy phosphates were depleted and tissue lactate content increased to 15 mumol X g-1. The total CO2 content (Tco2) was minimally or moderately reduced (normal, 11.9 mumol X g-1; range of ischemic values, 7.9-12.1 mumol X g-1), this range probably reflecting variable amounts of remaining blood flow. Impedance measurements demonstrated that ECF volume during ischemia was reduced to 55% of control, with gradual normalization during the first 15-30 min of recirculation. From values for Ptco2, Tco2, [HCO3-]e, and ECF volume, [HCO3-]i and pHi could be calculated. These values pertain to an idealized homogeneous intracellular compartment, and the methods used cannot detect whether different intracellular compartments diverge in their acid-base responses.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
123.
124.
Udo Wienand Ulrike Weydemann Ursula Niesbach-Klösgen Peter A. Peterson Heinz Saedler 《Molecular & general genetics : MGG》1986,203(2):202-207
Summary The c2 locus of Zea mays, identified as one of the genes affecting anthocyanin biosynthesis, was cloned using the transposable element En (Spm) as a gene tag. The Spm element present at the c2 locus in the autonomously mutating c2-m1 line was isolated using En1 element specific probes. Sequences flanking the element were identified as c2 locus specific and were used to clone the nonautonomous c2-m2 and wild-type alleles. The cloning and analysis of a cDNA complementary to the c2 locus provided evidence that this gene encodes the enzyme chalcone synthase. 相似文献
125.
O. L. Lomovskaya S. Z. Mindlin Zh. M. Gorlenko R. B. Khesin 《Molecular & general genetics : MGG》1986,202(2):286-290
Summary HgCl2-resistant strains of Acinetobacter sp. obtained from the soil at the Khaidarkan mercury mine (Kirghiz SSR) were found to contain, apart from large plasmids (60 kb), a small plasmid (7.5 kb) designated pKL1. It was established by conjugative crosses and transformation that pKL1 is a broad host range mobilizable plasmid and that it carries the Hgr determinant. The restriction map of pKL1 was constructed; the site of the Hgr determinant and the regions essential to replication were localized. A comparison of these results with earlier data suggests that microorganisms belonging to one microbiocenosis may carry Hgr determinants on plasmids with highly different structures and properties.Deceased on July 16, 1985 相似文献
126.
E. Wada R. Imaizumi Y. Kabaya T. Yasuda T. Kanamori G. Saito A. Nishimune 《Plant and Soil》1986,93(2):269-286
Summary Plants from agricultural and natural upland ecosystem were investigated for15N content to evaluate the role of symbiotic N2-fixation in the nitrogen nutrition of soybean. Increased yields and lower δ15N values of nodulating soybeansvs, non-nodulating isolines gave semi-quantitative estimates of N2 fixation. A fairly large discrepancy was found between estimations by δ15N and by N yield at 0 kg N/ha of fertilizer. More precise estimates were made by following changes in plant δ15N when fertilizer δ15N was varied near15N natural abundance level. Clearcut linear relationships between δ15N values of whole plants and of fertilizer were obtained at 30 kg N/ha of fertilizer for three kinds of soils. In experimental
field plots, nodulating soybeans obtained 13±1% of their nitrogen from fertilizer, 66±8% from N2 fixation and 21±10% from soil nitrogen in Andosol brown soil; 30%, 16% and 54% in Andosol black soil; 7%, 77% and 16% in
Alluvial soil, respectively. These values for N2 fixation coincided with each corresponding estimation by N yield method.
Other results include: 1)15N content in upland soils and plants was variable, and may reflect differences in the mode of mineralization of soil organics,
and 2) nitrogen isotopic discrimination during fertilizer uptake (δ15N of plant minus fertilizer) ranged from −2.2 to +4.9‰ at 0–30 kg N/ha of fertilizer, depending on soil type and plant species.
The proposed method can accurately and relatively simply establish the importance of symbiotic nitrogen fixation for soybeans
growing in agricultural settings. 相似文献
127.
Summary Non-symbiotic heterotrophic N2 fixation in coniferous bark litter was investigated with the acetylene reduction assay under aerobic and anaerobic conditions. The litter studied was composed essentially of bark, of pH 5 and a C/N ratio of 101; the ratio of available C to available N, which governs N2 fixation, was considerably higher. The rate of N2 fixation was estimated as 2.5–4.4 g N. g–1 dry wt. day–1. Nitrogenase activity was still evident after seven months of incubation under aerobic conditions. The N2-ase activity was O2 dependent: under anaerobic conditions no N2-ase activity was found unless a fermentable C source was added. The importance of N2 fixation in N-poor litter for the maintenance of soil fertility is emphasized. 相似文献
128.
[3H]Piflutixol binding to rat striatal membrane preparations identifies both D-1 and D-2 sites. We used [3H]piflutixol to characterise those binding sites present in 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS)-solubilised rat striatal preparations. The specific binding of [3H]piflutixol, as defined using cis-flupenthixol, to CHAPS-solubilised rat striatal tissue was saturable and of high affinity. Specific [3H]piflutixol binding to the solubilised preparations was displaced stereoselectively by the isomers of butaclamol and to an equal extent by both cis-flupenthixol and (+/-)-sulpiride. A positive correlation was found between the capacity of a range of drugs to displace [3H]piflutixol binding and the displacement of [3H]spiperone to the same preparations. The Bmax of [3H]piflutixol binding was not different from that of [3H]spiperone binding to the same preparation. These studies suggest that, in contrast to specific binding of membrane preparations, the specific binding of [3H]piflutixol to CHAPS-solubilised preparations involves mainly D-2 sites. Specific [3H]piflutixol binding, in contrast to [3H]spiperone binding, showed only slow dissociation from soluble preparations. The binding of [3H]piflutixol to CHAPS-solubilised preparations was retained during passage through a gel filtration column. This prelabelling of solubilised striatal preparations using [3H]piflutixol may aid in the purification of CHAPS-solubilised rat striatal D-2 sites. 相似文献
129.
Solubilization and Characterization of D2 -Dopamine Receptors in an Estrone-Induced, Prolactin-Secreting Rat Pituitary Adenoma 总被引:3,自引:2,他引:1
C. Bouvier M. Potier G. Beauregard J. Lafond N. Amlaiky† M. G. Caron† R. Collu 《Journal of neurochemistry》1986,47(5):1653-1660
D2-dopamine (3,4-dihydroxyphenylethylamine) receptors were successfully solubilized with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate from an estrone-induced rat pituitary adenoma. Forty-five percent of initial protein and 48% of initial [3H]spiroperidol binding sites were solubilized. The high affinity as well as the stereoselectivity of the sites was preserved. The order of potency of dopaminergic agonists was found to be typical of D2 receptors. Target size analysis by radiation inactivation indicated a molecular weight of 143,000 +/- 3,000 and of 106,000 +/- 4,000 daltons for membrane-bound and solubilized receptors, respectively. This suggests the loss of a 37,000-dalton subunit during solubilization without significant modification of binding characteristics. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of receptor protein preparation photolabeled with N-(p-azido-m[125I]iodophenethyl)spiroperidol confirmed the existence of a 94,000-dalton peptide which probably constitutes the ligand binding site of the receptor. Thus, our data indicate that chronic estrogen treatment of rats, although inducing a pituitary adenoma, does not modify the pharmacological characteristics of D2 receptors. These data suggest therefore that these adenoma may represent an ideal source of material for further biochemical characterization of D2 receptors. 相似文献
130.
Stimulation and inhibition of secretion by phorbol myristate acetate in different cell types 总被引:1,自引:0,他引:1
G J Blackwell R W Bonser J Dawson L G Garland 《Biochemical and biophysical research communications》1985,127(3):950-955
In washed human platelets and in HL60 granulocytes phorbol myristate acetate (PMA, 1-2000nM) synergised with threshold concentrations of secretogogues to induce a sustained maximum secretory response. Likewise, superoxide production from HL60 cells maintained a maximal response at PMA concentrations between 30-300nM. At concentrations up to 10nM PMA also augmented calcium ionophore, A23187, stimulated histamine release from rat peritoneal mast cells. However, in the mast cell PMA concentrations above 10nM reduced maximum histamine release in a dose-dependent manner. 相似文献