Photoaffinity Labeling with Progesterone-11α-Hemisuccinate-(2-[125I]Iodohistamine) Identifies Four Protein Bands in Mouse Brain Membranes

Abstract: The radiolabeled progesterone (PG) analogue progesterone-11α-hemisuccinate-(2-[¹²⁵I]iodohistamine) was used to label PG binding proteins in brain membranes from mouse cerebellum. Photoaffinity labeling and sodium dodecyl sulfate-polyacrylamide gel electrophoresis identified specific PG binding protein bands 1–4 of 64−29 kDa. Bands 1 and 4 were well resolved on the gel and easily quantified. Preincubation with PG inhibited photolabeling in a dose-dependent manner. The labeling was specific with respect to steroid structure. For band 1, the extent of inhibition of labeling by PG and 3α,5α-pregnanolone (3α) was pronounced. Other steroids such as testosterone (Tes), estradiol (Est), and corticosterone (Cor) were less effective, whereas pregnenolone sulfate (PS) and cholesterol (Cho) were ineffective. With respect to band 4, Est was the most effective; PG, 3α, and Tes were intermediate; and PS, Cho, and Cor were ineffective. The results describe specific membrane proteins that bind PG (band 1) and Est (band 4).     

Variations in stream water uptake by Eucalyptus camaldulensis with differing access to stream water

The stable isotopes ²H and ¹⁸O were used to determine the water sources of Eucalyptus camaldulensis at three sites with varying exposure to stream water, all underlain by moderately saline groundwater. Water uptake patterns were a function of the long-term availability of surface water. Trees with permanent access to a stream used some stream water at all times. However, water from soils or the water table commonly made up 50% of these trees' water. Trees beside an ephemeral stream had access to the stream 40–50% of the time (depending on the level of the stream). No more than 30% of the water they used was stream water when it was available. However, stream water use did not vary greatly whether the trees had access to the stream for 2 weeks or 10 months prior to sampling. Trees at the third site only had access to surface water during a flood. These trees did not change their uptake patterns during 2 months inundation compared with dry times, so were not utilising the low-salinity flood water. Pre-dawn leaf water potentials and leaf ¹³C measurements showed that the trees with permanent access to the stream experienced lower water stress and had lower water use efficiencies than trees at the least frequently flooded site. The trees beside the ephemeral stream appeared to change their water use efficiency in response to the availability of surface water; it was similar to the perennial-stream trees when stream water was available and higher at other times. Despite causing water stress, uptake of soil water and groundwater would be advantageous to E. camaldulensis in this semi-arid area, as it would provide the trees with a supply of nutrients and a reliable source of water. E. camaldulensis at the study site may not be as vulnerable to changes in stream flow and water quality as previously thought.     

The binding of zinc,but not cadmium,by phytic acid in roots of crop plants

Plant species adapted to soils enriched with heavy metals often accumulate these metals in their above or below ground organs. In this study, electron probe microanalysis of fractured, quench-frozen root specimens of common crop species shows that an appreciable quantity of Zn can be bound as Zn phytate (myo-inositolkis-hexaphosphate) within small vacuoles of cells in the root elongation zone of lucerne, soybean, lupins, tomato, rapeseed, cabbage, radish, maize and wheat exposed to high levels of Zn (80–300 M). Globular deposits of Zn phytate are most frequently observed in the endodermis of dicotyledonous species and in the pericycle of monocotyledonous species, but may also occur in the stele and inner cortex after prolonged exposure to toxic levels of Zn. The deposits could not be found in Zn-treated sunflower, field peas and Italian ryegrass. In three crop species, lucerne, soybean and maize, Zn-induced phytate globules were frequent, but exposure of roots to 30 M Cd did not induce the formation of Cd-containing globular deposits as observed inLemna minor (Van Steveninck et al., 1990a, 1992). Simultaneous Zn and Cd treatment induced the formation of Zn phytate globules as effectively as Zn alone, and Cd was not detected in the deposits.     

Chromium-induced inhibition of ethylene evolution in bean (Phaseolus vulgaris) leaves

The influence of chromium concentration on ethylene production in bean plants ( Phaseolus vulgaris L. cv. Contender) was investigated. A Cr ion-induced inhibition of ethylene synthesis from endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) was observed within both leaf discs floated on 2 m M CrO²⁻₄ or Cr³⁺ and leaf discs from plants cultured in nutrient solutions containing 10, 20 or 40 μ M CrO²⁻₄. However, Cr ions supplied either to plants with the nutrient solution or to discs with the incubation medium rather increased the conversion of exogenous ACC to ethylene. Primary leaves of plants exposed to CrO²⁻₄-containing nutrient solutions showed a statistically insignificant decrease of ACC-synthase activity. In the trifoliolate leaves of plants exposed to 10 μ M CrO²⁻₄, in which a significant decrease of ethylene production from endogenous ACC was observed, a substantial increase of ACC synthase was found. These results indicate that Cr ion-induced inhibition of ethylene production is not due to a breakdown of membrane integrity, which is necessary for ethylene forming enzyme activity, but caused by metabolic alterations leading to decreased ACC availability. Chromium ions may act by inhibiting ACC synthase activity or by diverting a metabolic step prior to the ACC synthase catalyzed reaction.     

Genotypic variation in biological nitrogen fixation by common bean

Field experiments were performed in Austria, Brazil, Chile, Colombia, Guatemala, Mexico and Peru as part of an FAO/IAEA Co-ordinated Research Programme to investigate the nitrogen fixing potential of cultivars and breeding lines of common bean (Phaseolus vulgaris L.). Each experiment included approximately 20 bean genotypes which were compared using the ¹⁵N isotope dilution method. Great differences in nitrogen fixation were observed between and within experiments, with average values of 35% N derived from atmosphere (% Ndfa) and highest values of 70% Ndfa being observed. These values which were larger than had been reported previously for common bean, were observed only when environmental factors were favorable. Therefore, common bean lines are available, which can support high biological nitrogen fixation. These can be used either directly as cultivars for production or in breeding programmes to enhance nitrogen fixation in other cultivars.     

Nitrogen relations of the sorghum-Sfriga hermonthica hostparasite association: growth and photosynthesis

The extent to which the parasitic angiosperm Striga hermonthica reduces the growth of its sorghum host is dependent on the concentration of nitrogen (as NH₄NO₃ in 40% Long Ashton Solution) supplied to the plants. The biomass of 0.5,1 and 2 mol m^?3 N-grown infected plants was 22,30 and 66%, respectively, of uninfected plants after 140d growth. The biomass of 3 and 4 mol m^?3 N-grown infected plants differed little from uninfected plants. No grain was set in 0.5 and 1 mol m^?3 N-grown infected plants, grain yield reached 42 and 73% of controls in 2 and 3 mol m^?3 N-grown plants, and was unaffected in 4 mol m^?3 N-grown plants. Striga hermonthica also altered the allometry and architecture of the host, at all but the highest N concentration. Higher N concentration (3 and 4 mol m ^?3 N) reduced the growth of S. hermonthica. Foliar N concentrations in sorghum ranged from 11 mg g^?1 dwt. in 0.5 mol m^?3 N-grown plants, to 28 mg g^?1 dwt. in 4 mol m^?3 N-grown plants, and were not affected by S. hermonthica. Higher N concentrations were measured in S. hermonthica, and ranged from 18 to 45 mg g^?1 dwt. in 0.5 and 3 mol m^?3 N-grown plants, respectively. The relationship between photosynthesis (CO₂ flux) and N concentration differed between uninfected and infected sorghum. This was most apparent in 0.5 mol m^?3 N-grown plants, with rates of 16 and 11 μmol m^?2 s^?1 in uninfected and infected plants, respectively (at 1500–1800 μmol m^?2 s^?1 photosynthetic photon flux density). At higher N concentrations, this difference was smaller, with both sets of plants reaching 26 μmol m^?2 s^?1 at 4 mol m^?3 N. Varying the level of S. hermonthica infection showed that the effect of N on host photosynthesis cannot be explained by differences in the mass or number of parasites supported by the host. At low levels of infection in 1 mol m^?3 N-grown plants, the negative effect of the parasite was reversed, and photosynthesis in infected plants exceeded that in uninfected plants by 20%. Photosynthesis in S. hermonthica at 3 mol m^?3 N (8 μmol m^?2 s^?1) was double that in 0.5 mol m^?3 N-grown plants. Stable carbon isotope and gas exchange measurements data demonstrated that this higher level of autotrophic carbon fixation was accompanied by a lower dependency on hetero trophic carbon. The latter ranged from 27 to 6% in 0 5 mol m^?3 and 3 mol m^?3 N-grown plants, respectively.     

Phytochelatin-mediated cadmium tolerance inschizosaccharomyces pombe

Summary Plants and certain fungi respond to heavy metal toxicity with the induced synthesis of metal-binding peptides known as phytochelatins (PCs). With cadmium, PCs can bind the metal to form a low molecular weight PC-Cd complex and a high molecular weight PC-Cd-S²⁻ complex. The sulfide ions enhance the stability and Cd-binding capacity of the metal chelate, and formation of this sulfide-containing complex is associated with enhanced tolerance to cadmium. Molecular analyses of two fission yeast mutants that fail to produce a wild type level of the PC-Cd-S²⁻ complex have determined that a vacuolar membrane transporter and several enzymes of the purine biosynthesis pathway are necessary in vivo for formation of the PC- Cd-S²⁻ complex. A model based on vacuolar sequestration of the PC-Cd complex by an ATP-binding cassette-type transporter and its subsequent maturation into the stable PC-Cd-S²⁻ complex via the actions of two purine biosynthetic enzymes is described. Presented in the Session-in-Depth Bioremediation through Biotechnological Means at the 1993 Congress on Cell and Tissue Culture, San Diego, CA, June 5–9, 1993.     

Proteomics in Drosophila melanogaster.

To be able to understand cellular mechanisms, we require fully integrated data sets combining information about gene expression, protein expression, post-translational modification states, sub-cellular location and complex formation. Proteomics is a very powerful technique that can be applied to interrogate changes at the protein level. Studying this effectively requires specialised facilities within research institutes. Here, we describe the setting up and operation of such a facility, providing a resource for the Arabidopsis and Drosophila research communities.