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101.
Summary The pattern of fatty acid release from rat synaptic membranes in the presence of phospholipase A2 (Vipera russelli) was compared to that from liposomes comprised of phospholipids. Phospholipase A2 more readily attacked myelin and synaptic membranes than liposomes prepared from total phospholipids derived from myelin. Although hydrolysis of liposomal phospholipids occurred in the absence of added calcium, the presence of 2mm CaCl2 or 2% bovine serum albumin significantly enhanced the phospholipase attack of liposomes, but not synaptic membranes or myelin. Phospholipase exhibited a marked preference for phospholipids containing docosahexaenoic acid (226) in the synaptic membranes, while with liposomes the pattern of released fatty acid reflected the fatty acid composition in the two-position of the phospholipids. Although either calcium or albumin markedly increased the phospholipase hydrolysis of liposomes, neither affected the hydrolysis of synaptic membranes or the pattern of fatty acid release from liposomes. It was concluded that the nonlipid constituents, particularly the proteins, of biomembranes were responsible for the organization of the phospholipids and accounted for the observed differences between liposomes and synaptic membranes with respect to enzymic accessibility. 相似文献
102.
Phytochrome was localized by immunoelectron microscopy in cells of the coleoptile tip of etiolated and irradiated oat (Avena sativa L., cv. Konata) seedlings. By using ultrathin frozen sections and immunopurified, monospecific antibodies, both the sensitivity and resolution of the immunocytochemical assay were increased. The results with etiolated plants agree with and extend previously published data. A brief red light illumination caused the redistribution of phytochrome from a diffuse to a more particulate appearance. Areas that accumulated phytochrome were identified as small vacuoles into which phytochrome was sequestered following illumination. In seedlings illuminated for several hours and in normal light-grown plants, the cellular distribution of phytochrome is qualitatively similar to that of nonirradiated, dark-grown material, except that in green plants the nucleus shows a positive immunocytochemical reaction. 相似文献
103.
The electronic structure of Fe2+ in reaction centers from Rhodopseudomonas sphaeroides. I. Static magnetization measurements. 总被引:2,自引:0,他引:2
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W F Butler D C Johnston H B Shore D R Fredkin M Y Okamura G Feher 《Biophysical journal》1980,32(3):967-992
We have measured the static magnetization of unreduced and reduced reaction centers that vary in their quinone content. Measurements were performed in the temperature range 0.7 degrees K less than T less than 200 degrees K and magnetic fields of up to 10 kG. The electronic g-value, crystal field parameters D, E, and the exchange interaction, J, between the quinone spin and Fe2+ were determined using the spin Hamiltonian formalism. The effective moment mu eff/Fe2+ of both reduced and unreduced samples were determined to be 5.35 +/- 0.15 Bohr magnetons. This shows, in agreement with previous findings, that Fe2+ does not change its valence state when the reaction centers are reduced. Typical values of D congruent to +5 cm-1 and E/D congruent to 0.27 are consistent with Fe being in an octahedral environment with rhombic distortion. The values of D and E were approximately the same for reaction centers having one and two quinones. These findings imply that quinone is most likely not a ligand of Fe. The Fe2+ and the spin on the quinone in reduced reaction centers were found to be coupled with an exchange interaction 0 less than /J/ less than 1 cm-1. The validity of the spin Hamiltonian was checked by using an orbital Hamiltonian to calculate energy levels of the 25 states of the S = 2, L = 2 manifold and comparing the magnetization of the lowest five states with those obtained from the spin Hamiltonian. Using the orbital Hamiltonian, we calculated the position of the first excited quintet state to be 340 cm-1 above the ground state quintet. This is in good agreement with the temperature dependence of the quadrupole splitting as determined by Mossbauer spectroscopy. 相似文献
104.
105.
Wayne K. Hoffman Peter Lalley Jean Deb Butler Sheldon Orloff Joseph D. Schulman Anil B. Mukherjee 《In vitro cellular & developmental biology. Plant》1981,17(8):735-740
Summary Using lipochromosomes (phospholipid-entrapped chromosomes) we have transferred the human HGPRT gene into HGPRT deficient mouse
cells (A9) with a frequency of approximately 1×10−5 (Mukherjee et al., Proc. Natl. Acad. Sci. USA 75: 1361–1365; 1978). Two other genes located on the long arm of the human
X-chromosome were also expressed in two independently derived populations of transferents (A9/GT3 and A9/GT4). We report here
the chromosomal and enzymatic composition of human HGPRT-positive clones from each subpopulation analyzed in detail with alkaline
Giemsa-11 staining.
All the clones expressed human PGK and HGPRT, but one (A9/GT4C6) lacked human G6PD. In each of four clones examined microscopically,
a small piece of presumptive human chromatin was visible in the karyotypes of most cells. The chromatin fragment was free
or attached in each cell of an individual clone. When integrated, the human chromosomal fragment in each clone appeared associated
with the centromere of the same telocentric A9 chromosome (No. 6 by Q-banding).
These data suggest that: (a) substantial human chromosomal fragments can be transferred into recipient cells using the lipochromosome
technique; (b) clones from human HGPRT positive A9 transferent subpopulations may or may not possess other human X-linked
markers; (c) the stability of lipochromosomally transferred genes varied from clone to clone and stability is generally poor
in the absence of continuous selection pressure (e.g., HAT); (d) when multiple X-linked human genes were transferred to mouse
cells a cytologically detectable human chromosomal fragment was identified free or attached to a host chromosome; and (e)
integration of transferred human chromosomal material into mouse chromosomes may occur at preferential site(s) in the recipient
genome.
This research was sponsored in part by the Office of Health and Environmental Research U.S. Department of Energy under Contract
W-7405-eng-26 with the Union Carbide Corporation. 相似文献
106.
Lung volume and pleural pressure effects on ventricular function 总被引:1,自引:0,他引:1
107.
Structure of transcriptionally-active chromatin subunits. 总被引:4,自引:0,他引:4
Rat liver chromatin is organized into regions of DNA which differ in degree of susceptibility to attack by the endonucleases DNase I and DNase II. The most nuclease-sensitive portion of chromatin DNA is enriched in transcribed sequences. This fraction may be separated from the bulk of chromatin by virtue of its solubility in solutions containing 2 mM MgCl2. Both transcribed and nontranscribed regions of chromatin are organized into repeating units of DNA and histone, which appear as 100 A beads in the electron microscope. The length of DNA in the repeat unit is the same for these two classes of chromatin (198 +/- 6 base pairs in rat liver); however, the subunits of active, Mg++-soluble chromatin differ from the nucleosomes of inactive regions of chromatin in several respects. Active subunits are enriched in nascent RNA and nonhistone protein and exhibit higher sedimentation values than the corresponding subunits of inactive chromatin. 相似文献
108.
James P. Butler 《Bulletin of mathematical biology》1977,39(5):543-563
The method of Green's functions is applied to the convection-diffusion equation describing gas transport in the human lung.
Explicit analytic solutions are found for the zeroth approximation to the value of steady state alveolar oxygen concentration, given the frequency of breathing, tidal volume,
oxygen consumption, alveolar volume, volume of the bronchial tree, and two modelling parameters describing the geometry. The
results of the calculation are compared favorably with some experimental data. 相似文献
109.
Reassembly of tobacco mosaic virus from the isolated RNA and protein, supplied as a disk preparation consisting of over 75% as the disk aggregate, has been followed by the protection of the RNA from nuclease digestion. The sizes of the RNA fragments were determined on agarose/acrylamide gels.During the first few minutes the protected RNA is found to be “quantized” into discrete lengths, differing on average by about 50 or 100 nucleotides, corresponding to one or two turns of the virus helix and strongly supporting the hypothesis that elongation in the major direction, towards the 5′-hydroxyl end, is occurring by the direct addition of protein disks. Protected RNA of the full length found in tobacco mosaic virus is visible within six minutes of starting reassembly, and by 30 minutes most of the RNA is fully protected. 相似文献
110.
Low temperature spectral properties of subchloroplast fractions purified from spinach 总被引:10,自引:8,他引:2
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Spinach (Spinacia oleracea L.) chloroplasts solubilized by digitonin were separated into five fractions by sucrose density gradient centrifugation. Three of the fractions, FI, FII, and FIII, corresponding to photosystem I, photosystem II, and the chlorophyll a/b complex, were purified further by two steps of diethylaminoethyl-cellulose chromatography followed by electrofocusing on an Ampholine column. The polypeptide patterns of the fractions were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the spectral properties of the fractions at −196 C determined by absorption spectra, fourth derivative curves of the absorption spectra, fluorescence emission spectra, and fluorescence excitation spectra. The activity of purified FII (photosystem II) was also assayed by the photoreduction of dichlorophenol-indophenol at room temperature using 1,5-diphenylcarbohydrazine as the electron donor and by the photoreduction of C-550 at −196 C. The different fractions showed unique polypeptide patterns and unique sets of low temperature-absorbing forms of chlorophyll. The fluorescence emission spectra of FI, FII, and FIII at −196 C were also unique with maxima at 734, 685 and 681 nm, respectively. FI showed negligible emission at wavelengths shorter than 700 nm and the long wavelength tails of FII and FIII in the 730 nm region were relatively small (approximately 10% of emission of their wavelength maxima). Addition of 0.1% Triton to FI and FII caused the longer wavelength absorbing forms of chlorophyll to shift to 670 nm and the fluorescence emission maxima (of both fractions) to shift to 679 nm at −196 C with an increase in the yield of fluorescence especially in the case of FI. 相似文献