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  • Molecular Cell Biology  (103)
  • Wiley-Blackwell  (103)
  • Periodicals Archive Online (PAO)
  • Springer
  • Springer Nature
  • 1975-1979  (103)
  • 1978  (103)
Collection
Publisher
  • Wiley-Blackwell  (103)
  • Periodicals Archive Online (PAO)
  • Springer
  • Springer Nature
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  • 1975-1979  (103)
Year
  • 1
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 9 (1978), S. 351-361 
    ISSN: 0091-7419
    Keywords: nerve growth factor ; receptors ; sensory ganglia cells ; brain cells ; serological receptor assay ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: When single-cell suspensions prepared from embroyonic day 8 (E8) chick sensory ganglia are incubated with nerve growth factor (NGF), anti-NGF antiserum, and complement, an NGF-dependent cytotoxic kill of 20 (±3)% of the ganglia cells is observed. This percentage is increased by a factor of two when only the neuronal cells are tested. No kill is observed on the nonneuronal cell population representing 50% of the ganglia dissociate. When E8 sensory ganglia cells are cultured in the presence of NGF following cytotoxic kill, the large, phase-bright NGF-reponsive neurons are missing from the culture. These results indicate that the cells recognized in the cytotoxicity assay have to carry NGF-binding sites of type I, which is the one with the higher affinity of the two types of NGF-binding sites (I and II) present on sensory ganglia cells. This conclusion is further supported by the following data: (a) half maximal cytotoxicity is reached already at a concentration of NGF which is below the KD of binding site I; (b) a washing step which removes all NGF bound to type II receptors while leaving a high percentage of type I receptors occupied has no effect on the percentage of ganglia cells killed.Using the cytotoxicity assay the presence of high-affinity binding sites of type I can be demonstrated on sensory ganglia cells from E8 chick embryos but not from E4 embryos and not on liver and heart cells from E8 embryos. Further, type I receptor-bearing cells were detectable in the brain using this assay. At E8, NGF receptors could be detected on cells of the forebrain and the tectum but not on brain stem cells. Cytotoxic kill of forebrain cells was found to be especially high at E8 and E9, and decreased by E10.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 9 (1978), S. 399-406 
    ISSN: 0091-7419
    Keywords: photoreactive probes ; ESR spin labels ; membranes ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: To investigate the dynamics of membrane processes that may be integral components of specific transmembrane signaling events we have synthesized several novel paramagnetic probes and their photoreactive counterparts. The structure of these probes was designed to (1) restrict “flipping” across the membrane bilayer; (2) contain paramagnetic or photoreactive moieties that could be placed at specific depths within the bilayer; (3) provide information about membrane structure as well as dynamics of protein movement; and (4) in the case of the photoreactive probes, be of high specific radioactivity.The molecules described in this paper consist of amino acid, dipeptide, or carbohydrate groups attached to arylazide- or nitroxide-bearing fatty acids. The synthesis and initial characterization of these membrane probes is described.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 9 (1978) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 4
    ISSN: 0091-7419
    Keywords: nuclear envelope-chromatin relationship ; chromosomes ; micronuclei ; mitochondria ; Colcemid ; EDTA and EGTA ; calcium magnesium ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: In the presence of the spindle poison Colcemid in the culture medium to prevent anaphase, approximately 20% of Chinese hamster metaphase cells were converted to micronucleated cells during 7 h. In the micronuclei the chromosomes had become enclosed by a nuclear envelope (NE). In the light-microscope the micronuclei were of two kinds: with either visible chromatids or with decondensed chromosomes. In the electron microscope (EM) the spatial relationship of the NE to the chromatin was of two kinds only in the presence of Colcemid. In about 90% of the micronucleated cells the spatial relationship was normal, ie, the NE was immediately adjacent to the chromatin. In the remaining cells, the NE was distended so that the outer NE was separated from the inner one. In the presence of the drivalent cation chelator, (ethylenedinitrilo) tetraacetic acid (EDTA) or the Ca2+-chelator [ethylenebis (oxyethylenenitrilo)] tetraacetic acid (EGTA), in addition to Colcemid, the amount of cells with micronuclei increased to 40%. The light-microscope appearance was the same as that found in the absence of the chelating agents. However, after Colcemid plus EGTA, EM revealed that only about 50% of the micronucleated cells had NE that was immediately adjacent to the chromatin and about 10% of them had distended outer NE. In the remaining 40% a third kind of spatial relationship was seen: the NE was intact but most of it was not adjacent to the chromatin. Furthermore, this type of micronucleus often contained mitochondria within the confines of NE. Thus, Ca2+ and possibly Mg2+ may regulate the rate of formation of the NE and also its ultrastructural relation to the chromatin. Mitochondrial function also appears to be involved in this relationship. In the presence of chloramphenicol (CAP), an inhibitor of mitochondrial protein synthesis, in addition to Colcemid, only about 50% of the micronucleated cells exhibited the normal relationship. The outer NE was separated from the inner NE in about 46% of the micronucleated cells and the third kind of NE-chromatin relationship was observed only in 2%. In the case of the third kind of relationship produced by CAP, inclusion of mitochondria within the micronuclei was not observed, in contrast to the finding with EGTA.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 9 (1978), S. 537-554 
    ISSN: 0091-7419
    Keywords: irreversibly sickled cells ; freeze-etching ; scanning electron micrography ; membrane-bound hemoglobin ; membrane proteins and glycoproteins ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Irreversibly sickled cells (ISCs) are sickle erythrocytes which retain bipolar enlongated shapes despite reoxygenation and owe their biophysical abnormalities to acquired membrane alterations. Freeze-etched membranes both of ISCs produced in vitro and ISCs isolated in vivo reveal microbodies fixed to the internal (PS) surface which obscure spectrin filaments. Intramembranous particles (IMPs) on the intramembrane (PF) surface aggregate over regions of subsurface microbodies. Electron microscopy of diaminobenzidine-treated ISC ghosts show the microbodies to contain hemoglobin and/or hemoglobin derivatives. Scanning electron microscopy and freeze-etching demonstrate that membrane-hemoglobin S interaction in ISCs enhances the membrane loss by microspherulation. Membrane-bound hemoglobin is five times greater in in vivo ISCs than non-ISCs, and increases during ISC production, paralleling depletion of adenosine triphosphate. Polyacrylamide gel electrophoresis of ISC membranes shows the presence of high-molecular-weight heteropolymers in the pre-band 1 region, a decrease in band 4.1 and an increase in bands 7, 8, and globin. The role of cross-linked membrane protein polymers in the generation of ISCs is discussed and is synthesized in terms of a unified concept for the determinants of the genesis of ISCs.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 8 (1978), S. 39-49 
    ISSN: 0091-7419
    Keywords: affinity chromatography ; plasma membrane ; neoplastic transformation ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The probe 2,4,6-trinitrobenzene sodium sulfonate may be used under appropriate conditions for selective labelling of plasma membrane proteins exposed at the outer cell surface. Labeled proteins, solubilized by detergents, can be purified by reverse immunoadsorption using antiprobe antibodies covalently linked to Sepharose 4B. This method has been applied to an investigation of the outer cell surface structure of chicken embryo and hamster fibroblasts. Coelectrophoresis in sodium dodecyl sulfate-polyacrylamide gels of probe-labeled membrane proteins purified from baby hamster kidney fibroblasts have shown that 7 major protein groups of different molecular weight are exposed on both control and Rous sarcoma or polyoma virus-transformed cells. Moreover, the transformed cells display a nonvirion component of 80-100 k daltons that is not labeled by the probe in normal cells. In fibroblasts transformed by a temperature sensitive Rous sarcoma virus mutant, that transforms at 37°C but not at 41°C, the expression of this component is related to the expression of the transformed phenotype.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 8 (1978), S. 129-138 
    ISSN: 0091-7419
    Keywords: freeze-fracturing ; membranes ; lipid phase separations ; B stearothermophilus ; temperature adaptation ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Bacillus stearothermophilus cells vary the lipid fatty acid composition of cytoplasmic membranes with growth temperature. Spin label studies of such membranes have been interpreted to indicate lateral lipid phase separations at the growth temperature. We have now used freeze-fracture electron microscopy to confirm the spin label studies. Freeze-fracture faces of protoplasts indicate slight but distinct protein aggregation at the growth temperature. Aggregation increases rapidly with decreasing quench temperature in wild-type cells. In contrast we were unable to demonstrate extended protein segregation in membranes of a temperature-sensitive mutant that contains more than 58% branched fatty acids.Storage of protoplasts for prolonged times below the lipid phase transition results in the appearance of corrugated fracture faces with 300- to 500-Å repeat patterns, although this organism does not synthesize lecithins.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 8 (1978), S. 177-190 
    ISSN: 0091-7419
    Keywords: fish melanophores ; electron microscopy ; microtubules ; tubulin ; quantitative analysis ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Isolated melanophores of the angelfish, Pterophyllum scalare, have been used in a morphometric analysis and a quantitative study of their microtubule system. Using transverse sections spaced at regular intervals, the changes associated with the process of pigment aggregation have been determined. Upon the concentration of pigment granules in the central cell region, almost half of the cytoplasmic portion is also withdrawn from the peripheral cell regions. Counts of microtubules within a cell sector in cells with pigment aggregated and dispersed, respectively, reveal (a) a constancy of the number of microtubules in this sector regardless of the distance from the cell center, and (b) a reduction of microtubule number in cells with pigment aggregated by about 58%. On the basis of these counts, the total number of microtubules has been calculated. In the dispersed state, about 2,400 microtubules extend between the center and the periphery of the cell, while their number is about 1,000 in the aggregated state.Using a 13-protofilament model of a microtubule and relevant data on size and molecular weight of microtubule subunits, the amount of tubulin present as microtubules is calculated. In the average, the cells contain 1.95·108 monomers corresponding to 1.78·10-8 mg tubulin. A tentative estimation of the concentration of tubulin inside a melanophore yields values of 6.1 mg/ml for the whole cell and 16.5 mg/ml for the cytoplasm alone (excluding membrane-bound organelles). Based on this estimation, a comparison, with microtubule assembly in vitro is made.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 8 (1978), S. 191-213 
    ISSN: 0091-7419
    Keywords: amino-phospholipids ; chemical probes ; red cell membrane ; valinomycin ; ion transport ; membrane topology ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The red cell membrane has an asymmetric arrangement of phospholipids. The amino-phospholipids are localized primarily on the inner surface of the membrane and the choline phospholipids are localized to a large extent on the outer surface of the membrane. Evidence is presented based on the use of covalent chemical probes in sequence that the red cell membrane contains heterogeneous domains of PE and PS and that the transport systems for Pi and K+ are asymmetrically arranged. Certain amino groups of PE, PS, and/or protein localized on the outer membrane surface are involved in Pi transport and certain amino groups of PE, PS, and/or protein localized on the inner surface of the membrane are involved in K+ transport.Cross-linking studies with DFDNB show that the cross-linked PE-PE molecules are rich in plasmalogens. This suggests that clusters of plasmalogen forms of PE occur in the membrane. Both PE and PS are cross-linked to membrane protein. These PE and PS molecules contain 24-28% 16:0 and 18:0 fatty acids and 12% fatty aldehydes. PE and PS molecules are cross-linked to a spectrin-rich fraction. It is proposed that the binding of spectrin to membrane PE and PS may help anchor spectrin to the inner surface of the membrane and regulate shape changes in the cell.K+-valinomycin forms a complex with TNBS and converts it from a non-penetrating proble to a penetrating probe. Valinomycin enhances K+ leak and Pi leak in the red cells. SITS inhibits completely the valinomycin-induced Pi leak and inhibits partially the valinomycin induced K+ leak. Valinomycin and IAA have additive effects on Pi leak. Ouabin has no effect on basal or valino-mycin-induced Pi leak. These data suggest that Pi leak and K+ leak occur by separate transport systems.In summary, the amino-phospholipids in the red cell membrane are asymmetrically arranged; some occur in clusters and some are closely associated with membrane proteins. Amino-phospholipids also are believed to bind spectrin to the inner surface of the membrane and also may play a role in cation and anion leak.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 8 (1978), S. 215-221 
    ISSN: 0091-7419
    Keywords: spectrin ; erythrocyte membrane ; membrane attachment site ; membrane protein mobility ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Interactions between spectrin and the inner surface of the human erythrocyte membrane have been implicated in the control of lateral mobility of the integral membrane proteins. We report here that incubation of “leaky” erythrocytes with a water-soluble proteolytic fragment containing the membrane attachment site for spectrin achieves a selective and controlled dissociation of spectrin from the membrane, and increases the rate of lateral mobility of fluorescein isothiocyanate-labeled integral membrane proteins (〉 70% of label in band 3 and PAS-1). Mobility of membrane proteins is measured as an increase in the percentage of uniformly fluorescent cells with time after fusion of fluorescent with nonfluorescent erythrocytes by Sendai virus. The cells are permeable to macromolecules since virus-fused erythrocytes lose most of their hemoglobin. The membrane attachment site for spectrin has been solubilized by limited proteolysis of inside-out erythrocyte vesicles and has been purified (V). Bennett, J Biol Chem 253:2292 (1978). This 72,000-dalton fragment binds to spectrin in solution, competitively inhibits association of 32P-spectrin with inside-out vesicles with a Ki of 10-7M, and causes rapid dissociation of 32P-spectrin from vesicles. Both acid-treated 72,000-dalton fragment and the 45,000 dalton-cytoplasmic portion of band 3, which also was isolated from the proteolytic digest, have no effect on spectrin binding, release, or membrane protein mobility. The enhancement of membrane protein lateral mobility by the same polypeptide that inhibits binding of spectrin to inverted vesicles and displaces spectrin from these vesicles provides direct evidence that the interaction of spectrin with protein components in the membrane restricts the lateral mobility of integral membrane proteins in the erythrocyte.
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