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  • 1990-1994  (5)
  • 1970-1974  (1)
  • ATPase  (2)
  • ion channels  (2)
  • Atomic, Molecular and Optical Physics  (1)
  • Cell & Developmental Biology
  • Pisum sativum
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  • 1
    ISSN: 1432-2048
    Keywords: ATPase ; Plasma membrane ; Pyrophosphatase ; Ricinus (solute transport) ; Sucrose transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A highly enriched plasma membrane fraction has been isolated from dark-grown cotyledons ofRicinus communis by phase partitioning. This is demonstrated by the properties of the associated ATPase: high vanadate sensitivity, azide and nitrate insensitivity, sharp pH optimum around 6.5, and high specificity for ATP as substrate. The upper plasma membrane fraction also contained a pyrophosphatase activity, normally considered to be located on the tonoplast or Golgi membranes, which showed a specific activity higher than that in the lower phase. Sucrose gradient centrifugation of both microsomal and upper phase fractions showed a comigration of some pyrophosphatase activity with the plasma membrane fraction. Sucrose uptake changes with development inRicinus cotyledons. The ATPase activity in the upper (plasma membrane) phase also varied in a similar way with development, whereas activity in the lower phase showed little change. Pyrophosphatase activity in the upper phase also increased with development but did not show a peak and fall as seen for sucrose uptake and ATPase. The possibility that changes in plasma membrane ATPase may contribute to changes in sucrose uptake capacity and the possible cellular origin and physiological significance of the pyrophosphatase activity are discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1424
    Keywords: ion channels ; phosphorylation ; modulation of ion channels ; lens fibers ; reconstitution ; intercellular junctions ; major intrinsic protein ; gap junctions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Major intrinsic polypeptide (MIP), a 28-kDa protein isolated from lens fiber cell membranes, forms large, nonselective channels when reconstituted into lipid bilayers. MIP channels are regulated by voltage, such that these channels close when the potential across the membrane is greater than 30 mV. We have investigated the modulation of the voltage-dependent closure of MIP channels by phosphorylation. In this report, we describe the isolation of two isomers of MIP from lens fiber cell membranes. These isomers differ by a single phosphate at a protein kinase A phosphorylation site. The phosphorylated isomer produces channels that close in response to applied voltages when reconstituted into bilayers. The nonphosphorylated isomer produces voltage-independent hannels. Direct phosphorylation with protein kinase A converts voltage-independent channels to voltage-dependent channels in situ. Analyses of macroscopic and single channel currents suggest that phosphorylation increases the voltage-dependent closure of MIP channels by increasing closed channel lifetimes and the rate of channel closure following the application of voltage.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 128 (1992), S. 91-102 
    ISSN: 1432-1424
    Keywords: lens ; embryonic ; gap junctions ; ion channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Ion channels are believed to play an important role in the maintenance of lens transparency. In order to ascribe junctional and nonjunctional permeability properties to specific lens cell types, embryonic chick lenses were enzymatically dissociated into cell clusters, cell pairs and single cells, and both cell-to-cell and single-membrane permeability properties were characterized with the patch-clamp technique. Double patch-clamp experiments and single patch-clamp experiments with Lucifer yellow in the pipette demonstrated that the cells in the dissociated preparation were well coupled, the average conductance between pairs being 42 ± 27 nS. Double patch-clamp experiments also revealed single cell-to-cell channel events with a predominant unitary conductance of 286 ± 38 pS. Whole-cell measurements of surface membrane conductance indicate heterogeneity within the population of dissociated embryonic chick lens cells: 63% of the cells have a voltage-independent leak current, 14% of the cells have a potassium-selective inward rectifier current, and 23% of the cells have a current which turns off with positive voltage on a time scale on the order of seconds. The time constant for this turnoff is voltage dependent.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 165 (1991), S. 27-36 
    ISSN: 1615-6102
    Keywords: ATPase ; Cerium ; Cytochemistry ; Fixation ; Zea mays roots ; Plasma membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cytochemical localization of ATPase activity has been investigated in maize root cells using both lead and cerium-based capture methods. With both methods, staining at the plasma membrane was observed in all cells of the root, although the precipitate obtained with cerium was more uniform and granular than that with lead. Controls using no substrate or no magnesium, β-glycerophosphate to replace ATP, vanadate or boiled tissue generally showed little or no staining. However, biochemical studies on purified plasma membrane fractions showed that ATPase activity was markedly inhibited by fixation, particularly by glutaraldehyde, and also by lead and cerium ions. Non-enzymic hydrolysis of ATP by cerium was greater than that by lead. The value and limitations of these procedures for the localization of plasma membrane H+-ATPase activity are summarized in relation to previous criticisms of these methods.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    International Journal of Quantum Chemistry 38 (1990), S. 85-92 
    ISSN: 0020-7608
    Keywords: Computational Chemistry and Molecular Modeling ; Atomic, Molecular and Optical Physics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: Ab initio SCF were performed on the trans and gauche rotamers of succinonitrile (SN) and on the monohydrated rotamers to determine the most probable hydration site(s) of water with SN and the relative strengths of these interactions and to examine the effect of one water molecule on the relative stability of the rotamers. MOPAC was used to determine the location of water on the rotamers of SN; the geometries of the hydrated rotamers were then optimized at the HF/4-31G level followed by single-point calculations at the HF/6-31G and HF/6-31G* levels. The results show that trans SN is more stable than is gauche SN by 1.2 kcal/mol (at the HF/4-31G level with zero-point energy corrections) and 0.8 kcal/mol (at the HF/6-31G* level, excluding zero-point energy corrections). The hydrogen bond formed between the methylene hydrogen of trans SN and the oxygen of water is more stable than is the hydrogen bond formed between the nitrogen of SNt and the hydrogen of water by 0.63 kcal/mol. The most stable hydrogen-bonding interaction for the gauche rotamer occurs for the methylene - OH2 hydrogen-bonding interaction, which also has the largest hydration energy (ΔE = -4.39 kcal/mol), but it is less stable than is the most stable trans hydrogen-bonding interaction (SNt - OH2) by 0.63 kcal/mol. Our results suggest that the trans rotamer of SN should associate with more water molecules than should the gauche. The relative rotamer stability between the trans and gauche rotamers of SN decreases by 0.2 kcal/mol (at the HF/6-31G* level) when water is hydrogen bonded to the methylene group of SN. A mechanism is also proposed to explain the phase-separation behavior of this system.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 180 (1974), S. 597-603 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The minimum period of uterine exposure required by ejaculated boar spermatozoa as a preliminary to rapid capacitation has been determined after natural or surgical deposition of sperm samples directly into the uterine lumen. Twenty-four oestrous gilts were mated or inseminated close to the time of ovulation, and 15, 30, 45 or 60 minutes later, the Fallopian tubes were separated from the uterine cornua. The tubes were flushed at pre-arranged intervals during a second intervention, and the proportion of eggs penetrated and activated examined by phase-contrast microscopy.On the basis of 166 eggs recovered from eighteen mated gilts, a period of uterine exposure as brief as 30 minutes, when followed by a tubal residence of approximately three hours, permitted 30.3% of the eggs to be activated; this proportion increased to 51.6% and 60.5% if the tubes were isolated 45 or 60 minutes, respectively, after mating (p 〈 0.001), as did the mean number of spermatozoa associated with the eggs. When the cornua were separated from the tubes 15 minutes after semen deposition into the uterus of six animals, 11.3% of 62 eggs were fertilized during the ensuing three and one half hours, but very few spermatozoa had reached and/or attached to the eggs in this group.It is concluded that a population of boar spermatozoa potentially capable of effecting fertilization may enter the tubes within 15 to 30 minutes of mating near the time of ovulation, and that such vanguard spermatozoa can activate a proportion of the eggs within a further two to three hours. Thus, from a temporal point of view, the major components of the capacitation process in oestrous pigs are inferred to take place in the Fallopian tubes.
    Additional Material: 1 Tab.
    Type of Medium: Electronic Resource
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