Protein kinase activity and substrates at the surface of intact HeLa cells

Evidence is presented for the location at the surface of HeLa cells of a protein kinase capable of phosphorylating surface as well as extracellular (foreign) proteins. The reaction products have been found to be proteins containing phosphoryl groups as monoesters of seryl and threonyl residues (but not of tyrosine). The enzyme is of the cyclic AMP-independent type, since neither cyclic AMP nor the heat- and acid-stable inhibitor protein (specific for cyclic AMP-dependent protein kinases) influenced its activity. Further, co-substrate ATP could in part be substituted by GTP, and the spectrum of proteins phosphorylated by the ecto-enzyme differed from that phosphorylated by cyclic AMP-dependent protein kinases. Evidence for the ecto-enzymic nature of this protein kinase includes (a) utilization of co-substrate and location of products at the surface of cells carefully controlled as being in an intact state and (b) phosphorylation of exogenous protein (phosvitin; specific serum proteins) by intact cells. Conclusive proof was gained by qualitative and quantitative comparative studies of phosphorylation in cultures with varying degrees of damaged cells either as a whole or after separation into groups of intact and damaged cells by electronic cell sorting. The results of experiments with cell sonicates excluded the possibility that either enzyme or substrates released from damaged cells were simply adsorbing to the cell surface.     

Neurophysiology of the vibration sense in locusts and bushcrickets: Response characteristics of single receptor units

The response characteristics of the vibration receptors in the legs of the migratory locust, Locusta migratoria, and the tettigoniid Decticus verrucivorus were investigated electro-physiologically by single cell recordings. The legs were stimulated by sinusoidal vibrations. There are four types of vibration receptor in each leg of Locusta and Decticus, which can be classified physiologically. One type—most probably campaniform sensilla—shows a phase-locked response to vibrations from 30 to 200 Hz, its threshold reflecting the displacement. A second type shows similar responses in the same frequency range, but its reactions depend on the stimulus acceleration. The receptor cells of the subgenual organ are very sensitive to vibration from 30 to at least 5000 Hz, and their responses depend on acceleration. There are two types of subgenual receptors, one of which shows a clear maximum of sensitivity between 200 and 1000 Hz, with a threshold below 0.01 m/sec^?2 acceleration. Subgenual receptors with different thresholds and different characteristic frequencies occur in each leg. The receptors of each leg pair have quite similar mean sensitivities and characteristic frequencies. However, in the front legs of tettigoniids the more sensitive subgenual receptors and an additional receptor type also respond to low-frequency airborne sound up to 10 kHz.     

Isolation and identification of spirorenone metabolites from the monkey (Macaca fascicularis)

The plasma level of spirorenone was determined 3 h after 1, 8, 22 and 46 daily oral administrations of 20 mg/kg to two female and two male monkeys ($\text{Macaca fascicularis}$). A fifth animal, female, was treated with eight daily doses of tritium-labelled drug and was completely bled from the carotid vein 4 h after the last administration in order to isolate and identify plasma metabolites.After repeated daily doses of spirorenone the mean plasma level of unchanged drug was 711 ± 213 ng/ml. In the plasma of the fifth animal four radioactively labelled compounds could be detected after extraction and subsequent HPLC separation. Mass spectrometric identification of three of the substances indicated 1,2-dihydrospirorenone, hydroxy-1,2-dihydrospirorenone and the unchanged drug itself.     

Autonomic nervous control of gastric somatostatin secretion from the perfused rat stomach

The effect of parasympathetic and sympathetic nerve stimulation on the secretion of gastric somatostatin and gastrin has been studied in an isolated perfused rat stomach preparation. Stimulation of the vagus nerve inhibited somatostatin secretion and increased gastrin release. Splanchnic nerve stimulation increased somatostatin release during simultaneous atropine perfusion, but not in its absence, whereas gastrin secretion was unchanged. The secretory activity of the gastric D-cell was therefore reciprocally influenced by the sympathetic and parasympathetic nerves but sympathetic stimulation was only effective during muscarinic blockade.     

Hirsutinolides from Vernonia species

Out of the nineteen species of Vernonia studied, five contained highly oxygenated sesquiterpene lactones, while the rest contained predominantly triterpenes, especially lupane derivatives.     

Eudesmanolides and diterpenes from Wedelia trilobata and an ent-kaurenic acid derivative from Aspilia parvifolia

The reinvestigation of the aerial parts of Wedelia trilobata afforded, in addition to known compounds, six new eudesmanolides, two ent-kaurenic acid derivatives as well as a degraded one. A corresponding hydroxy compound was present in Gnaphalium undulatum. From Aspilia parvifolia new ent-kaurenic acid epoxides were isolated. The structures were elucidated by spectroscopic methods. The chemotaxonomic situation is discussed briefly.     

O(3)-(2-Acetylamino-2-deoxy-β-d-glucopyranosyl)-oleanolic acid,a novel triterpenoid glycoside from two Pithecellobium species

A new triterpenoid glycoside containing an amino sugar moiety has been isolated from Pithecellobium cubense and P. arboreum and identified as O(3)- (2-acetylamino-2-deoxy-β-d-glucopyranosyl)oleanolic acid. β-d-Glucopyranosyl-α-spinasterol was also obtained from P. cubense.     

Chlorophyll biosynthesis by mesophyll protoplasts and plastids from etiolated oat (Avena sativa L.) leaves

The uptake of [1-³H]geranylgeranyl diphosphate (GGPP) into protoplasts and intact etioplasts and the metabolic interconversion therein was studied after a 2 min pulse of white light. The chlorophyll synthetase reaction, Chlide+GGPPChl_GG, was taken as a natural probe for the etioplast compartment. This reaction yields labeled ChL_GG and, by hydrogenation, labeled Chl_P, when [1-³H]GGPP receives access to the etioplast stroma. It was found that penetration across the plastid envelope was rapid and that penetration across the plasma membrane of protoplasts, however, was slow. A cellular pool of soluble GGPP was detected. This pool was lost, in part, during preparation of the protoplasts and almost completely during preparation of the etioplasts. The membrane-bound phytol pool of etioplasts could not be replaced by exogenous [³H]GG. The endogenous GG and phytol pools of protoplasts, which were larger than those of etioplasts, could be replaced in part by exogenous [³H]GGPP. That part of this pool exists as soluble GGPP or as a direct precursor in the cytoplasm is discussed.Abbreviations GGPP geranylgeranyldiphosphate - Chl_GG geranylgeranyl chlorophyllide a - Chl_P phytyl chlorophyllide a - IPP isopentenyl diphosphate - Chlide chlorophyllide a     

Turgor pressure and water transport properties of suspension-cultured cells of Chenopodium rubrum L.

The turgor pressure and water relation parameters were determined in single photoautotrophically grown suspension cells and in individual cells of intact leaves of Chenopodium rubrum using the miniaturized pressure probe. The stationary turgor pressure in suspension-cultured cells was in the range of betwen 3 and 5 bar. From the turgor pressure relaxation process, induced either hydrostatically (by means of the pressure probe) or osmotically, the halftime of water exchange was estimated to be 20±10 s. No polarity was observed for both ex- and endosmotic water flow. The volumetric elastic modulus, , determined from measurements of turgor pressure changes, and the corresponding changes in the fractional cell volume was determined to be in the range of between 20 and 50 bar. increases with increasing turgor pressure as observed for other higher plant and algal cells. The hydraulic conductivity, Lp, is calculated to be about 0,5–2·10^–6 cm s^–1 bar^–1. Similar results were obtained for individual leaf cells of Ch. rubrum. Suspension cells immobilized in a cross-linked matrix of alginate (6 to 8% w/w) revealed the same values for the half-time of water exchange and for the hydraulic conductivity, Lp, provided that the turgor pressure relaxation process was generated hydrostatically by means of the pressure probe. Thus, it can be concluded that the unstirred layer from the immobilized matrix has no effect on the calculation of Lp from the turgor pressure relaxation process, using the water transport equation derived for a single cell surrounded by a large external volume. By analogy, this also holds true for Lp-values derived from turgor pressure changes generated by the pressure probe in a single cell within the leaf tissue. The fair similarity between the Lp-values measured in mesophyll cells in situ and mesophyll-like suspension cells suggests that the water transport relations of a cell within a leaf are not fundamentally different from those measured in a single cell.