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41.
Charles M. Lent Joyce Ono Kent T. Keyser Harvey J. Karten 《Journal of neurochemistry》1979,32(5):1559-1563
Abstract— We have measured serotonin (5-HT) within large and small neurosomata which are vitally stained by Neutral Red dye. A micro-radioenzymatic technique which is sensitive to 50fmol of 5-HT was employed on intact ganglia, 75 μm Retzius Cells (RZ) and a 10 μm ventro-lateral cell (VL) taken from the leech Macrobdella decora. The stain does not affect the levels of 5-HT in either ganglia or RZ. The VL cell body contains 5-HT at concentrations of at least 100 m m . Microspectrofluorometry of all the ganglionic neurosomata which fluoresce following the Falck-Hillarp formaldehyde condensation reaction detected rapidly-fading emission peaks of 509–523 nanometers. We conclude that all seven fluorescent neurons in the leech ganglion very probably contain serotonin. 相似文献
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The possible inhibition of adenylyl cyclase activity by atrial peptides selective for the ANF-C receptor was investigated in bovine pulmonary artery endothelial cells. In these cells isoprenaline, guanine nucleotide and forskolin dose-dependently increased activity over basal levels. In the presence of rANF(99-126), these dose-dependent increases were not reduced, nor were they affected by the ANF-C receptor selective analogue C-ANF(102-121). Furthermore, the selective analogues rANF(103-123) and des[Cys105,Cys121]rANF104-126 had no effect on basal or stimulated adenylyl cyclase activity. It can be concluded that ANF-C receptors are not linked to inhibition of adenylyl cyclase in these cells. 相似文献
45.
Phosphorylation of CTP:phosphocholine cytidylyltransferase in vivo. Lack of effect of phorbol ester treatment in HeLa cells 总被引:1,自引:0,他引:1
The production and characterization of an antibody to rat liver CTP:phosphocholine cytidylyltransferase is described. This antibody quantitatively precipitated cytidylyltransferase from both rat liver and HeLa cell cytosol. Following affinity purification, the antibody was used to demonstrate, for the first time, the phosphorylation of cytidylyltransferase in vivo. Following the immunoprecipitation of cytidylyltransferase from HeLa cells, acid hydrolysis, and thin layer electrophoresis of the amino acids, only [32P]phosphoserine was detected. The phosphorylation state of cytidylyltransferase in HeLa cells was examined following treatment with phorbol ester for 1 h. In agreement with previous studies, the incorporation of [3H]choline into phosphatidylcholine via the CDP-choline pathway was stimulated 5-fold in cultures of HeLa cells following treatment with phorbol ester for 1 h. However, no appreciable translocation of cytidylyltransferase was detected, despite the utilization of two different methods of cell lysis. Furthermore, the inclusion of phosphatase inhibitors and chelators of divalent cations in the homogenization buffers had no effect on the observed distribution or activity of the enzyme. Immunoprecipitated cytidylyltransferase was phosphorylated to the same extent, and on serine residues only, in both control and 12-O-tetradecanoyl phorbol-13-acetate (TPA)-treated cells. Measurement of the pool sizes of the aqueous intermediates of the CDP-choline pathway, following TPA treatment, revealed a modest decrease in the phosphocholine pool only, consistent with an activation of cytidylyltransferase. 相似文献
46.
Two polyclonal antibodies were raised to synthetic peptides corresponding to amino acids Ser21-Tyr35 and Lys247-Phe261 of cytochrome b561. These antibodies were used to test the native orientation of the amino and carboxyl termini of this transmembrane electron transport protein. Carboxyl-terminal epitopes were lost when intact chromaffin granules were treated with Pronase. This result indicates that the carboxyl terminus is cytoplasmically exposed and confirms a theoretical prediction obtained from hydropathy plots. Epitopes that were recognized by an amino-terminal antipeptide antibody were not removed under the same conditions. This finding implied that the amino terminus was not proteolytically accessible on the exterior of the granule. The abundance of threonine and serine residues in the amino-terminal region suggested that the amino terminus could be held in the membrane by covalent fatty acylation. Treatment of purified delipidated cytochrome b561 with hydroxylamine resulted in the release of a fatty acid hydroxamate. Sulfhydryl analysis of purified cytochrome b561 showed that all 3 cysteine residues were in the free sulfhydryl form. These observations indicate that cytochrome b561 is covalently fatty acylated and that the lipid is bound through ester linkages of serine or threonine residues. 相似文献
47.
A. Henry Latorella Susan K. Bromberg Kent Lieber James Robinson 《Journal of phycology》1981,17(2):211-214
Fifteen nitrate assimilation-deficient mutants of the euryhaline green alga, Dunaliella tertiolecta Butcher were selected by their chlorate resistance. Ten mutants, unable to grow on NO3? but able to grow on NO2?, had no detectable nitrate reductase activity. Five mutants, unable to grow on either NO3? or NO2?, had depressed levels of both nitrate and nitrite reductase. A method for assaying methyl viologen-nitrate reductase in the presence of nitrite reductase is described. 相似文献
48.
Properties of the Photosynthetic System and DNA of Cyanophora paradoxa Cyanelles 总被引:1,自引:1,他引:0
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The cyanelle from the photosynthetic biflagellate protist Cyanophora paradoxa has been studied in terms of its photosynthetic properties. Structurally, the cyanelle resembles unicellular cyanobacteria. The cyanelle is readily released from the host cell by means of the French press. The isolated cyanelle shows typical photosystem I and photosystem II activities as well as phenazine methosulfate-mediated photophosphorylation. The kinetic parameters Km and Vmax were determined for CO2 fixation in the cyanelle and cells of C. paradoxa and compared to a cyanobacterium. The determined values were not much different, although the cyanobacterium had a significantly greater rate of CO2 fixation, and the cyanelle was least active in this regard. Photosystem I chlorophyll-protein complex is readily isolated from the thylakoid membrane. In all these respects, the photosynthetic apparatus of the cyanelle resembles that of cyanobacteria. No nitrogen fixation activity was observed. Attempts to regenerate the isolated cyanelle were not successful, but in some cases, an unidentified cyanobacterium grew up in standing cultures of C. paradoxa cyanelles. Buoyant density data indicate that the strain of C. paradoxa we have investigated differs from that employed by others, since our strain shows a value of 1.716 grams per cubic centimeter and others report values of 1.695 and 1.691. 相似文献
49.
Simultaneous kinetic analysis of ribulose 1,5-bisphosphate carboxylase/oxygenase activities 总被引:4,自引:4,他引:0
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An assay was developed for simultaneous kinetic analysis of the activities of the bifunctional plant enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase [EC 4.1.1.39]. [1-14C,5-3H]Ribulose 1,5-bisphosphate (RuBP) was used as the labeled substrate. Tritium enrichment of the doubly labeled 3-phosphoglycerate (3-PGA) product, common to both enzyme activities, may be used to calculate Vc/Vo ratios from the expression A/(B-A) where A and B represent the 3H/14C isotope ratios of doubly labeled RuBP and 3-PGA, and Vc and Vo represent the activities of carboxylase and oxygenase, respectively. Doubly labeled substrate was synthesized from [2-14C]glucose and [6-3H]glucose using the enzymes of the pentose phosphate pathway coupled with phosphoribulokinase. 相似文献
50.
Vidmantas A. Raisys Patrick N. Friel Patricia R. Graaff Kent E. Opheim Alan J. Wilensky 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1980,183(4)
A one-step method for extraction of diazepam, nordiazepam, and internal standard into toluene is followed by chromatographic separation and detection with either dual-wavelength high-performance liquid chromatography or electron-capture gas—liquid chromatography. Agreement between the two methods was excellent for diazepam (r = 0.99, n = 38) and good for nordiazepam (r = 0.96, n = 79) over a concentration range that included subtherapeutic, therapeutic, and toxic plasma levels. 相似文献