Comparison of the catalytic and structural properties of Ca2+-ATPase in longitudinal tubules and terminal cisternae of the sarcoplasmic reticulum

The catalytic behavior and structural features of Ca2+-ATPase in the vesicles of longitudinal tubules and terminal cisternae of the sarcoplasmic reticulum isolated from rabbit skeletal muscles was analysed. pH measurements have shown under optimal conditions Ca2+-ATPase has similar catalytic behavior both in the fractions of longitudinal tubules and terminal cisternae. Under non-optimal conditions, the behavior similarity was not observed. The specific activity of the ATPase enzyme under optimal conditions was shown to be much higher in the fraction of longitudinal tubules than in the fraction of terminal cisternae. Caffeine added to both fractions had no effect on the catalytic behavior of Ca2+-ATPase. As judged from fluorescence analysis, the structure of Ca2+-ATPase of longitudinal tubules differs from that structure of terminal cisternae. In sarcoplasmic reticulum membrane, at least half of the tryptophan residues of Ca2+-ATPase was shown to be buried in the lipid bilayer. Our findings suggest that in terminal cisternae some of the Ca2+-ATPase molecules exist as an oligomeric protein and do not participate in ATP hydrolysis (named "silent" Ca2+-ATPase).     

Antigenic determinants of the constant region of human immunoglobin kappa-chain detected by monoclonal antibodies

Ten different monoclonal antibody (monAB) preparations reacting with human IgL chains of the kappa type have been obtained. Nine of the monAB interacted with the kappa-chain C domain, whereas only one monAB reacted with the V domain. It has been determined that monAB against the C domain react with three different epitopes. One epitope is expressed on intact Ig molecules as well as on isolated kappa-chains, whereas the other two epitopes are found only on isolated kappa-chains. The expression of these epitopes in 40 different myeloma kappa-chain preparations belonging to four various subgroups was studied. The level of this C domain epitope expression has been shown to depend on the variable subgroups of kappa-chains indicating a close association between V and C domains. This association leads to the alteration of antigenic activity of some C domain epitopes. The alterations are thought to be local because, as a rule, they involve only one of the three epitopes.     

Cotranscription of the S10- and spc-like operons in spinach chloroplasts and identification of three of their gene products

The organisation and expression of the rpl22, rps3, rpl16 and rpl14 genes, which belong to the S10- and spc-like operons of spinach chloroplasts, have been studied. Northern experiments and nuclease S1 mapping show that the two operon-like groups of genes are cotranscribed. It is demonstrated that the intron-containing rpl16 gene is spliced in vivo. Based on amino acid composition and protein sequence data, the products of the rpl22, rpl16 and rpl14 genes are identified respectively as the spinach chloroplast ribosomal proteins CS-L13, CS-L24 and CS-L29. The rpl22 gene product is a 5S rRNA binding protein and therefore is distinguishable from the homologous Escherichia coli L22 ribosomal protein.     

The effect of initial bioenergetic state on the cryostability of yeast cells

The cryostability of Saccharomyces cerevisiae cells decreased when they were cultivated under anaerobic conditions in a liquid growth medium YEPD as compared to the culture grown under aerobic conditions. The effect of cultivation conditions on the different cryostability of S. cerevisiae cells is discussed. The initial state of their bioenergetics was shown to influence the cryostability of yeast cells.     

The study of the growth of tylosin producer using differential centrifugation of mycelium in a sucrose density gradient

The mycelium of Streptomyces fradiae was fractionated by differential centrifugation in a sucrose density gradient (SDG) using various samples of the inoculation material and aliquots of the cultural broth taken in the course of tylosin production. The mode of mycelium distribution in SDG made it possible to select the most active inoculation material. The mycelium was redistributed from sucrose layers with a high density to those with a lower density in the course of fermentation. The fractions differed in the antibiotic activity but none of them had an activity higher than in the control centrifuged in 30% sucrose and washed off just like the fractions. Therefore, mycelium fractionation in SDG would not elevate its antibiotic activity. The paper presents the cytological characteristics of different fractions changing in the course of fermentation.     

Sensitivity of Escherichia coli cell membranes to various classes of detergents

The mechanisms of interaction between non-ionic or cationic surfactants with Escherichia coli K-12 cell membranes were studied using an approach based on the registration of changes in the membrane permeability to ethidium bromide, a fluorescent dye for nucleic acids. Triton X-100, a non-ionic detergent, was shown to exert no effect on the permeability of intact cell membranes. Triton X-100 interacted with the bacteria only after treatment with EDTA, a complexing agent for bivalent cations. Cetyltrimethyl ammonium bromide increased the permeability to ethidium bromide and the action of this cationic detergent did not require the pretreatment with the complexing agent. SDS, an anionic detergent, damaged E. coli K-12 and this could be registered by the lowering of intensity of light scattering by the bacterial suspension. The surface charge of E. coli K-12 cells was shown to influence the interaction of ionic detergents with bacterial cell membranes. Its variation by changing the pH of the incubation medium did not make E. coli K-12 sensitive to Triton X-100.     

A comparison of soil climate and biological activity along an elevation gradient in the eastern Mojave Desert

Summary Soil temperature, moisture, and CO₂ were monitored at four sites along an elevation transect in the eastern Mojave Desert from January to October, 1987. Climate appeared to be the major factor controlling CO₂ partial pressures, primarily through its influence of rates of biological reactions, vegetation densities, and organic matter production. With increasing elevation, and increasing actual evapotranspiration, the organic C, plant density, and the CO₂ content of the soils increased. Between January and May, soil CO₂ concentrations at a given site were closely related to variations in soil temperature. In July and October, temperatures had little effect on CO₂, presumably due to low soil moisture levels. Up to 75% of litter placed in the field in March was lost by October whereas, for the 3 lower elevations, less than 10% of the litter placed in the field in April was lost through decomposition processes.     

Luxury consumption and specific utilization rates of three macroelements in two Taraxacum microspecies of contrasting mineral ecology

Plants of Taraxacum sellandii Dahlst., a microspecies adapted to fertile, and Taraxacum nordstedtii Dahlst., adapted to infertile soils, were cultured hydroponically, either on a complete nutrient solution or on one deprived of nitrogen, phosphorus, or potassium ions. For all four treatments, the growth and internal mineral concentration of the plants was monitored. For plants cultured on a complete nutrient solution, the uptake rates of nitrate, phosphate, and potassium ions were determined. Luxury consumption of the three macronutrients was computed as the excess of ion absorption over the ion uptake rates minimally required to sustain maximum growth. In these calculations the critical N, P, or K⁺ concentrations, earlier derived, were used as parameters describing the mineral status minimally required to allow maximum growth. Efficiency in use of the three macroelements at various levels of mineral accumulation was also computed. Finally, the response to phosphate starvation as related to phosphate uptake capacity and the accumulation of P was investigated.
The physiological properies investigated provide a causal background for the superior adaptation of T. nordstedtii as compared to T. sellandii to infertile sites. Taraxacum nordstedtii had a higher relative luxury consumption of NO₃^–, H₂PO^-₄, and K⁺, a higher efficiency in N and P use at N– and (severe) P-deficiency, respectively; and, after phosphate starvation, a relatively high preservation of phosphate uptake capacity and an enlargement of P storage. In combination with the low potential growth, luxury consumption will be particularly effective in T. nordstedtii in preventing or minimizing mineral deficiency. The distribution of minerals between cytoplasm and vacuoles as a factor in mineral use efficiency is discussed.     

Gas exchange and water balance of a mistletoe species and its mangrove hosts

Summary The gas exchange and water relations of the hemiparasite Pthirusa maritima and two its mangrove host species, Conocarpus erectus and Coccoloba uvifera, were studied in an intertidal zone of the Venezuelan coast. Carbon uptake and transpiration, leaf osmotic and total water potential, as well as nutrient content in the xylem sap and leaves of mistletoes and hosts were followed through the dry and wet season. In addition, carbon isotope ratios of leaf tissue were measured to further evaluate water use efficiency. Under similar light and humidity conditions, mistletoes had higher transpiration rates, lower leaf water potentials, and lower water use efficiencies than their hosts. Potassium content was much higher in mistletoes than in host leaves, but mineral nutrient content in the xylem sap of mistletoes was relatively low. The resistance of the liquid pathway from the soil to the leaf surface of mistletoes was larger than the total liquid flow resistance of host plants. Differences in the daily cycles of osmotic potential of the xylem sap also indicate the existence of a high resistance pathway along the vascular connection between the parasite pathway along the vascular connection between the parasite and its host. P. maritima mistletoes adjust to the different physiological characteristics of the host species which it parasitizes, thus ensuring an adequate water and carbon balance.     

Evidence for Chloroplastic Succinate Dehydrogenase Participating in the Chloroplastic Respiratory and Photosynthetic Electron Transport Chains of Chlamydomonas reinhardtii

A method for isolating intact chloroplasts from Chlamydomonas reinhardtii F-60 was developed from the Klein, Chen, Gibbs, Platt-Aloia procedure ([1983] Plant Physiol 72: 481-487). Protoplasts, generated by treatment with autolysine, were lysed with a solution of digitonin and fractionated on Percoll step gradients. The chloroplasts were assessed to be 90% intact (ferricyanide assay) and free from cytoplasmic contamination (NADP isocitrate dehydrogenase activity) and to range from 2 to 5% in mitochondrial contamination (cytochrome c oxidase activity). About 25% of the cellular succinate dehydrogenase activity (21.6 micromoles per milligram chlorophyll per hour, as determined enzymically) was placed within the chloroplast. Chloroplastic succinate dehydrogenase had a K_m for succinate of 0.55 millimolar and was associated with the thylakoidal material derived from the intact chloroplasts. This same thylakoidal material, with an enzymic assay of 21.6 micromoles per milligram chlorophyll per hour was able to initiate a light-dependent uptake of oxygen at a rate of 16.4 micromoles per milligram chlorophyll per hour when supplied with succinate and methyl viologen. Malonate was an apparent competitive inhibitor of this reaction. The succinate dehydrogenase activity present in the chloroplast was sufficient to account for the photoanaerobic rate of acetate dissimilation in H₂ adapted Chlamydomonas (M Gibbs, RP Gfeller, C Chen [1986] Plant Physiol 82: 160-166).