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81.
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83.
Dipeptidase and carboxypeptidase A activities were determined in cells and luminal contents of the fore-, mid-, and hind-midgut of Musca domestica larvae. Dipeptidase activity was found mainly in hind-midgut cells, whereas carboxy-peptidase activity was recovered in major amounts in both cells and in luminal contents of hind-midguts. The subcellular distribution of dipeptidase and part of the carboxypeptidase A activities is similar to that of a plasma membrane enzyme marker (aminopeptidase), suggesting that these activities are bound to the microvillar membranes. Soluble carboxypeptidase A seems to occur both bound to secretory vesicles and trapped in the cell glycocalyx. Based on density-gradient ultracentrifugation and thermal inactivation, there seems to be only one molecular species of each of the following enzymes (soluble in water or solubilized in Triton X-100): membrane-bound dipeptidase (pH optimum 8.0; Km 3.7 mM GlyLeu, Mr 111,000), soluble carboxypeptidase (pH optimum 8.0; Km 1.22 mM N-carbobenzoxy-glycyl-L-phenylalanine (ZGlyPhe), Mr45,000) and membrane-bound carboxypeptidase (pH optimum 7.5, Km 2.3 mM ZGlyPhe, Mr58,000). The results suggest that protein digestion is accomplished sequentially by luminal trypsin and luminal carboxypeptidase, by membrane-bound carboxypeptidase and aminopeptidase, and finally by membrane-bound dipeptidase. 相似文献
84.
Standing biomass and production in water drainages of the foothills of the Philip Smith Mountains, Alaska 总被引:1,自引:0,他引:1
Steven J. Hastings Scott A. Luchessa Walter C. Oechel John D. Tenhunen 《Ecography》1989,12(3):304-311
In the foothills of the Philip Smith Mountains, Brooks Range, Alaska, tussock tundra is the most widely distributed vegetation, and it occurs on rolling hills and in valleys that were shaped by a sequence of Pleistocene glaciations. In this study, aboveground standing biomass and production were compared in "intertrack tundra" areas that were relatively homogenous with respect to downslope drainage and adjacent "water tracks" that acted to channel water flow to the valley bottom stream. Comparisons of biomass, leaf area index, and specific leaf weight were also made between upper and lower slope positions. Similarities and differences of vegetation structure are examined with respect to graminoid, deciduous shrub, evergreen shrub, herbaceous, and bryophyte components.
Water tracks were found to have 1.5–1.7 times the biomass of intertrack tundra, and production (excluding secondary growth) in water tracks was 40% greater than in intertrack tundra. The aboveground biomass for all areas studied and the annual production values were similar to those found in other studies of tussock tundra. While only slight differences in depth of thaw occurred in water tracks and intertrack tundra during June and early July, water tracks thawed more deeply with the onset of summer rains. Warmer temperatures at 40 cm depth in July and August may have increased nutrient availability, whereas greater rooting depth and movement of water may have increased nutrient capture in water tracks as compared with the intertrack areas. Greater biomass and a deeper thaw depth occurred at upper slope locations. 相似文献
Water tracks were found to have 1.5–1.7 times the biomass of intertrack tundra, and production (excluding secondary growth) in water tracks was 40% greater than in intertrack tundra. The aboveground biomass for all areas studied and the annual production values were similar to those found in other studies of tussock tundra. While only slight differences in depth of thaw occurred in water tracks and intertrack tundra during June and early July, water tracks thawed more deeply with the onset of summer rains. Warmer temperatures at 40 cm depth in July and August may have increased nutrient availability, whereas greater rooting depth and movement of water may have increased nutrient capture in water tracks as compared with the intertrack areas. Greater biomass and a deeper thaw depth occurred at upper slope locations. 相似文献
85.
Structural basis for variations in the sensitivity of human decay accelerating factor to phosphatidylinositol-specific phospholipase C cleavage. 总被引:5,自引:0,他引:5
E I Walter W L Roberts T L Rosenberry W D Ratnoff M E Medof 《Journal of immunology (Baltimore, Md. : 1950)》1990,144(3):1030-1036
Human decay-accelerating factor (DAF) proteins expressed on E and nucleated cells differ in their susceptibility to phosphatidylinositol (PI)-specific phospholipase C (PLC) cleavage/release. To investigate the mechanism of this difference, the glycoinositol-phospholipid anchoring moieties of E DAF, and of HeLa cell, polymorphonuclear cell, and lymphocyte DAF were structurally compared. Labeling of PI-PLC-resistant E DAF with 3-(trifluoromethyl)-3-(m-[125I]-iodophenyl)-diazirine ([125I]TID) and TLC analysis of nitrous acid deamination anchor fragments showed a predominant phospholipid species with less polar migration than the 125I-TID-labeled PI. Gas chromatographic analyses of methanolyzed E protein revealed 2.20 +/- 0.16 mol of fatty acids [16:0, 18:0, 18:1, 20:4, 22:4, and 22:5 (0.76, 0.36, 0.25, 0.15, 0.40, 0.28 mol, respectively)] and 0.86 +/- 0.05 mol of inositol per mol of N-terminal Asp. Gas chromatography-mass spectroscopy demonstrated principally myo-inositol but also variable amounts of the chiro-isomer. Nondenaturing polyacrylamide gel electrophoresis of 14C-radiomethylated E protein revealed that pretreatment with hydroxylamine, a reagent which removes ester-linked lipids, rendered it PI-PLC susceptible. In contrast, parallel analyses of 35S-cys-labeled PI-PLC-sensitive HeLa DAF protein revealed only minor amounts of the hydroxylamine-sensitive phospholipid species. Similar results were obtained with 125I-surface-labeled DAF from polymorphonuclear cells, as well as from unstimulated peripheral blood and anti-CD3-activated lymphocytes. These findings demonstrate that, rather than PI, the E DAF anchor contains an inositol alkylacylglycerol-phospholipid which is heterogeneous with respect to acyl groups and inositol isomers, that an ester-linked substitution in this inositolphospholipid underlies the resistance of E DAF protein to PI-PLC cleavage/release, and that this structural modification is cell-specific. 相似文献
86.
87.
Fusion of cultured dog kidney (MDCK) cells: I. Technique,fate of plasma membranes and of cell nuclei
Ulrich Kersting Heribert Joha Wieland Steigner Birgit Gassner Gerhard Gstraunthaler Walter Pfaller Hans Oberleithner 《The Journal of membrane biology》1989,111(1):37-48
Summary The evaluation of the intracellular signal train and its regulatory function in controlling transepithelial transport with electrophysiological methods often requires intracellular measurements with microelectrodes. However, multiple impalements in epithelial cells are hampered by the small size of the cells. In an attempt to avoid these problems we fused cells of an established cell line, Madin Darby canine kidney cells, originally derived from dog kidney, to giant cells by applying a modified polyethylene glycol method. During trypsin-induced detachment from the ground of the petri dish, individual cells grown in a monolayer incorporate volume and mainly lose basolateral plasma membrane by extrusion. By isovolumetric cell-to-cell fusion, spherical giant cells are formed within 2 hr. During this process a major part of the individual cell plasma membranes is internalized. Over three weeks following cell plasma membrane fusion degradation of single cell nuclei and cell nuclear fusion occurs. We conclude that this experimental approach opens the possibility to investigate ion transport of epithelia in culture by somatic cell genetic techniques. 相似文献
88.
Release of recombinant microbes into the environment necessitates an evaluation of their ability to transfer genetic material. The present report evaluates a method to detect conjugal DNA plasmid transfer in soil slurries under various environmental conditions. DonorPseudomonas cepacia containing pR388::Tn1721 andP. cepacia recipient cultures were coincubated in soil slurries containing autoclaved or natural soil and treated with one or more of 14 experimental conditions. Conjugal mating frequency (transconjugants per initial donor) ranged from 4.8×10–1 to 1.9×10–7. Highest numbers of transconjugants, 1.5×107 colony forming units/ml soil slurry, were observed following incubation at 35°C with an enriched nutrient supplement added to the soil. Low numbers of transconjugants, 103 colony forming units/ml soil slurry, were observed when mating pairs were subjected to low nutrient or pH stress even though initial donor and recipient populations were maintained at high levels. This test system provides a simple way to estimate effects of changing environmental factors on plasmid transfer rates and on the survival of recombinant microorganisms. By use of soil collected from sites proposed to receive genetically engineered microorganisms, preliminary risk assessments can be obtained regarding the potential for gene transfer and microorganism survival with this soil slurry test system. 相似文献
89.
Evidence for a two-step mechanism involved in assembly of functional signal recognition particle receptor 总被引:13,自引:6,他引:7 下载免费PDF全文
The signal recognition particle (SRP) and SRP receptor act sequentially to target nascent secretory proteins to the membrane of the ER. The SRP receptor consists of two subunits, SR alpha and SR beta, both tightly associated with the ER membrane. To examine the biogenesis of the SRP receptor we have developed a cell-free assay system that reconstitutes SR alpha membrane assembly and permits both anchoring and functional properties to be assayed independently. Our experiments reveal a mechanism involving at least two distinct steps, targeting to the ER and anchoring of the targeted molecule on the cytoplasmic face of the membrane. Both steps can be reconstituted in vitro to restore translocation activity to ER microsomes inactivated by alkylation with N-ethyl-maleimide. The characteristics elucidated for this pathway distinguish it from SRP-dependent targeting of secretory proteins, SRP-independent ER translocation of proteins such as prepromellitin, and direct insertion mechanisms of the type exemplified by cytochrome b5. 相似文献
90.
The role of litter in an old-field community: impact of litter quantity in different seasons on plant species richness and abundance 总被引:10,自引:0,他引:10
Summary We studied the effect of removing and adding plant litter in different seasons on biomass, density, and species richness in a Solidago dominated old-field community in New Jersey, USA. We removed all the naturally accumulated plant litter in November (658 g/m2) and in May (856 g/m2) and doubled the amount of litter in November and May in replicated plots (1 m2). An equal number of plots were left as controls. Litter removal and addition had little impact on total plant biomass or individual species biomass in the growing season following the manipulations. Litter removal, however, significantly increased plant densities but this varied depending upon the season of litter removal, species, and life history type. Specifically, the fall litter removal had a much greater impact than the spring litter removal suggesting that litter has its greatest impact after plant senescence in the fall and prior to major periods of early plant growth in spring. Annual species showed the greatest response, especially early in the growing season. Both spring and fall litter removal significantly increased species richness throughout the study. Litter additions in both spring and fall reduced both plant densities and species richness in June, but these differences disappeared near the end of the growing season in September. We concluded than in productive communities where litter accumulation may be substantial, litter may promote low species richness and plant density. This explanation does not invoke resource competition for the decline in species richness. Finally, we hypothesize that there may be broad thresholds of litter accumulation in different community types that may act to either increase or decrease plant yield and diversity. 相似文献