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  • Articles  (22)
  • actin  (22)
  • Wiley-Blackwell  (22)
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1981), S. 179-192 
    ISSN: 0886-1544
    Keywords: actin ; echinoderm ; fascin ; filopodia ; actin cross-linking protein ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Echinoderm coelomocytes transform from petaloid cells with large motile lamellipodia to filopodial forms. During this morphological transformation, actin filaments extensively reorganize from a random meshwork into tight bundles, which become the skeletons or cores of the filopodia. Antibody localization procedures show that fascin, a 58,000 dalton actin cross-linking protein, becomes incorporated into the filament bundles as they form. Isolated filopodial cores have a pronounced transverse striping pattern, which has been previously identified with fascin crosslinks, and gel electrophoresis identifies a protein in the cores that co-migrates with purified egg fascin. A few of the core fragments also have a distinctive “cap,” which we presume is the membrane insertion site for actin filaments.We have developed a radioimmunoassay for fascin and have used it to study the redistribution of this protein during transformation. Data from the assay indicate that fascin constitutes about 5% of the total cell protein and that substantially more fascin, approximately 1.5-2 times more, is found in the Triton-insoluble cytoskeletons of the filopodial cells than in the petaloid cells. Actin, measured by the DNAase I inhibition assay accounts for approximately 10% of the total cell protein. Approximately 65% of this actin is in a soluble non-filamentous form in the petaloid cells. Our results show that actin polymerization must occur during the cell shape change, since we find approximately 25% more actin in the filopodial cytoskeleton than in the petaloid cytoskeleton. The results show a preferential incorporation of fascin into the cytoskeleton as the cells form filopodia.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 1 (1980), S. 31-40 
    ISSN: 0886-1544
    Keywords: actin ; fascin ; actin cross-linking proteins ; fertilization ; microvilli ; sea urchin eggs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Following fertilization, the sea urchin egg cortex undergoes a structural change involving the assembly and organization of actin filaments into microvilli. Antifascin localizes this actin cross-linking protein in the microvilli of the fertilized egg cortex but no organized staining is present in the unfertilized cortex. Determination of the actin content of eggs using the DNAase I inhibition assay indicates that actin is about 1.4% of the total protein. Approximately 90% of this actin is soluble in low calcium isotonic extracts of unfertilized eggs while only 60-65% can be recovered in identical extracts of fertilized eggs. Similar measurements for fascin using a radioimmunoassay indicate this molecule represents about 0.3% of the total egg protein, essentially all of which is recovered in low calcium isotonic extracts of unfertilized eggs. After fertilization only 65-70% of this actin cross-linking protein is in the soluble phase. These results demonstrate a markedly different solubility for actin and fascin after fertilization, when the indirect immunofluorescence staining localizes fascin in the microvilli, and are consistent with the idea that fascin organizes newly polymerized actin filaments into the microvillar cores. A consideration of the amounts of actin and fascin incorporated into the cortex after fertilization and the number of microvilli on the egg surface indicates that the measured values are sufficient to account for the observed microvillar elongation.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 13 (1989), S. 245-263 
    ISSN: 0886-1544
    Keywords: actin ; actin-binding protein ; plasma membranes ; cytoskeleton ; immunofluorescence microscopy ; cell motility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ponticulin is the major actin-binding integral glycoprotein in plasma membranes isolated from log-phase Dictyostelium discoideum amebae. As such, this protein appears to be an important link between the plasma membrane and actin filaments (Wuestehube and Luna: Journal of Cell Biology 105:1741-1751, 1987). In this study, indirect immunofluorescence microcopy was used to examine the distribution of ponticulin in randomly moving D. discoideum amebae and in amebae engaged in cell migration and phagocytosis. Ponticulin is distributed throughout the plasma membrane and also is present in intracellular vesicles associated with the microtubule-organizing center-Golgi complex adjacent to the nucleus. In aggreating amebae, ponticulin is concentrated in regions of lateral cell-cell contact and in arched regions of the plasma membrane. Ponticulin also is present, but not obviously enriched, in filopodia, in the actin-rich anterior end of polarized cells, and in detergent-insoluble cytoskeletons. In amebae engaged in phagocytosis of yeast, ponticulin is present but not enriched in phagocytic cups and is associated with intracellular vesicles around engulfed yeast. These results suggest that ponticulin is stably associated with actin filaments in certain regions of the plasma membrace and that the actin-binding activity of ponticulin may be tightly controlled.Indirect immupofluorescence microscopy and immunoblot analysis demonstrate that human polymorphonuclear leukocytes also contain a 17 kD protein that specifically cross-reacts with antibodies affinity-purified aganst D. discoideum ponticulin. As in D. discoideum, the mammalian 17 kD ponticulin-analog appears to be localized in plasma membrane and is evident in actin-rich cell extensions. These results indicate that ponticulin-mediated linkages between the plasma membrane and actin may be present in higher eukaryotic cells.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 25 (1993), S. 43-48 
    ISSN: 0886-1544
    Keywords: malaria ; Plasmodium falciparum ; merozoite ; actin ; ubiquitin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Merozoites of the human malaria parasite, Plasmodium falciparum, when treated with cytochalasin B, will attach irreversibly to red cells with formation of a vestigial internal (parasitophorous) vacuole, but they are inhibited from moving into the cell. The existence of an actin-based motile mechanism is implied. Immunoblotting, peptide mapping and the DNase inhibition assay have been used to show that the merozoite contains actin. It makes up an estimated 0.3% of the total parasite protein and is partitioned in the ratio of about 1:2 between the cytosolic and particulate protein fractions. In the former it is unpolymerised and in the latter filamentous. Most of the anti-actin-reactive protein in the soluble fraction and about 20% of that in the pellet has an apparent molecular weight of 55,000 and reacts with an anti-ubiquitin antibody; it is thus evidently ubiquitinyl actin, or arthrin, which has so far been detected only in insect flight muscle. © 1993 Wiley-Liss, Inc.
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  • 5
    ISSN: 0021-9304
    Keywords: surface topography ; actin ; vinculin ; fibronectin ; vitronectin ; grooves ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: The microfilaments and vinculin-containing attachment complexes of rat dermal fibroblasts (RDF) incubated on microtextured surfaces were investigated with confocal laser scanning microscopy (CLSM) and digital image analysis (DIA). In addition, depositions of bovine and endogenous fibronectin and vitronectin were studied. Smooth and microtextured silicone substrata were produced that possessed parallel surface grooves with a groove and ridge width of 2.0, 5.0, and 10.0 μm. The groove depth was approximately 0.5 μm. CLSM and DIA make it possible to visualize and analyze intracellular and extracellular proteins and the underlying surface simultaneously. It was observed that the microfilaments and vinculin aggregates of the RDFs on the 2.0 μm grooved substrata were oriented along the surface grooves after 1, 3, 5, and 7 days of incubation while these proteins were significantly less oriented on the 5.0 and 10.0 μm grooved surfaces. Vinculin was located mainly on the surface ridges on all textured surfaces. In contrast, bovine and endogenous fibronectin and vitronectin were oriented along the surface grooves on all textured surfaces. These proteins did not seem to be hindered by the surface grooves since many groove-spanning filaments were found on all the microgrooved surfaces. In conclusion, it can be said that microtextured surfaces influence the orientation of intracellular and extracellular proteins. Although results corroborate three earlier published hypotheses, they do not justify a specific choice of any one of these hypotheses. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 291-300, 1998.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Supramolecular Structure 9 (1978), S. 373-389 
    ISSN: 0091-7419
    Keywords: surface receptors ; capping ; endocytosis ; actin ; myosin ; Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have made observations, by double fluorescence staining of the same cell, of the distributions of surface receptors, and of intracellular actin and myosin, on cultured normal fibroblasts and other flat cells, and on lymphocytes and other rounded cells. The binding of multivalent ligands (a lectin or specific antibodies) to a cell surface receptor on flat cells clusters the cell receptors into small patches, which line up directly over the actin- and myosin-containing stress fibers inside the cell. Similar ligands binding to rounded cells can cause their surface receptors to be collected into caps on the surface, and these caps are invariably found to be associated with concentrations of actin and myosin under the capped membrane. Although these ligand-induced surface phenomena appear to be different on flat and rounded cells, we propose that in both cases clusters of receptors become linked across the membrane to actin- and myosin-containing structures. In flat cells these structures are very long stress fibers; therefore, when clusters of receptors become linked to these fibers, the clusters are immobilized. In round cells, membrane-associated actin- and myosin-containing structures are apparently much less extensive than in flat cells; therefore, clusters of receptors linked to these structures are still mobile in the plane of the membrane. We suggest that in this case the clusters are then actively collected into a cap by an analogue of the muscle sliding filament mechanism.To explain the transmembrane linkage, we propose that actin is associated with the plasma membrane as a peripheral protein which is directly or indirectly bound to an integral protein (or proteins) X of the membrane. Individual molecules of any receptor are not bound to X, but after they are specifically clustered into patches, a patch of receptors then becomes bound to S and hence to actin/myosin.Patching and capping of specific receptors on rounded cells is often accompanied by a specific endocytosis of the ligand-receptor complexes. This represents one common transport mechanism of a protein (the ligand) across the plasma membrane. The possibility is discussed that this type of endocytosis is mediated by a transmembrane linkage of the clustered receptor to actin/myosin. Another mechanism of endocytosis involves the “coated pit” structures that are observed by electron microscopy of plasma membranes. The possible relationships between an actin/myosin and a coated pit mechanism of endocytosis are explored.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 3 (1983), S. 525-534 
    ISSN: 0886-1544
    Keywords: actin ; actin-membrane interactions ; coelomocytes ; calmodulin ; cytoskeleton ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Coelomocytes from several echinoderm species undergo an actin-mediated cytoskeletal transformation once subjected to hypotonic shock. In this study, coelomocytes from the sea urchins Lytechinus variegatus and Arbacia punctulata were induced to “transform” by treatment with 〉 5 μM of the calcium ionophore A23187 in the presence of external Ca++. The dependence of ionophore transformation on external Ca++ and the lack of chlorotetracycline staining indicates that these cells rely on external Ca++ sources. NBD-phallacidin (7-Nitrobenz-2-oxa-1,3-diazole-phallacidin) staining of lysolecithin permeabilized cells and wholemount transmission electron microscopy (TEM) show that similar reorganizations of the actin cytoskeleton take place during hypotonic shock and ionophore transformation, although actin filament bundling is less apparent in A23187-treated cells. As has been shown with hypotonic shock transformation, the ionophore elicited shape change is inhibited by anticalmodulin drugs. Greater than 10 μM concentrations of W 13 inhibit filopod formation, while this drug's less active structural analogue, W 12, exhibits no effects. W 13 also appears to disrupt actin filament-membrane associations in the cells. Fluorescent localization of calmodulin using a photooxidized derivative of trifluoperazine indicates a general cytoplasmic distribution with some concentration in filopod core bundles. Coelomocyte transformation may be an example of a cellular shape change regulated by Ca++ through the action of calmodulin modulation of actin-membrane interactions.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 7 (1987), S. 347-360 
    ISSN: 0886-1544
    Keywords: cytochalasin ; actin ; microtubules ; immunofluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Indirect immunofluorescence demonstrated a dramatic reorganization of cytokeratin filaments produced by cytochalasin B (CB) treatment of PtK1 cells. Much of the normal cytokeratin network became arranged into a latticework consisting of bundles of cytokeratin filaments that radiated from, and interconnected, distinct foci, Electron microscopy showed foci to be dense granular regions through which bundles of cytokeratin filaments looped. Composition of the foci included actin, myosin, and alpha-actinin, as shown by labeling with rhodamine phalloidin or specific antisera. Simultaneous treatment with CB and colchicine was not required for lattice formation, but did produce more extensive development than did CB alone. In cells treated only with CB, the microtubule network remained intact, even in regions of extensive lattice formation. These results contrast sharply with those of Knapp et al (J. Cell Biol. 97:1788 [1983b]), who found lattice formation dependent upon simultaneous CB and colchicine treatment. Time-course and dose-response studies of CB treatment showed lattice formation to follow disruption of stress fibers and the concentration of actin into distinct patches that marked the location of lattice foci. Overall results suggest a structural association between microfilaments and cytokeratin filaments that produces the lattice pattern upon CB-induced disruption of stress fibers. Lattice formation was not limited to a specific cell-cycle stage, since G1, G2, and M cells displayed the lattice. Treatment of cells with dihydro-CB and experiments with enucleated cells showed that lattice formation was dependent upon neither the inhibition of sugar transport nor the nuclear extrusion effects of CB.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 11 (1988), S. 318-325 
    ISSN: 0886-1544
    Keywords: actin ; contractile proteins ; microvilli ; cytoskeleton ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The mammalian genome contains 20-30 genes encoding a family of actins. To date, however, only six proteins (four muscle and two nonmuscle isoforms) encoded by this multigene complex have been identified. We have isolated two actins from the brush border of rat intestinal epithelial cells that have isoelectric points and N-terminal peptides characteristic of the cytoplasmic β- and γ-actins. However, using a panel of actin-specific monoclonal antibodies, we show that these actins contain a set of epitopes that distinguishes them from any of the known cytoplasmic or muscle isoforms. These unique actins share features of both the nonmuscle and muscle isoforms, suggesting that they represent an intermediate in the evolution of the specialized muscle actins.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 12 (1989), S. 12-22 
    ISSN: 0886-1544
    Keywords: actin ; CDPK ; cytoskeleton ; cytochalasin D (CD) ; rhodamine-phalloidin (RP) ; pollen ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We recently purified a calcium-dependent but calmodulin- and phospholipid-independent protein kinase (CDPK) from cultured plant cells (Harmon et al.: Plant Physiology 83:830-837, 1987). A monoclonal antibody (mAb 3B9) directed against CDPK was used to localize this protein in Allium root cells and Tradescantia pollen tubes using immunofluorescence techniques. The mAb 3B9 staining pattern showed that CDPK is localized within a fibrous network in the cytoplasm resembling the normal interphase network of F-actin. Treatment of tissue with 10 μM cytochalasin D (CD) prior to fixation abolished the staining pattern. Double-localization experiments in which pollen tubes were first stained with mAb 3B9 and then with rhodamine-phalloidin (RP) demonstrated that CDPK and F-actin were colocalized. Monoclonal antibody 3B9 did not react with purified actin from rabbit muscle or Dictyostelium and did not bind to proteins corresponding to the Mr of actin in crude extracts of Allium root tips and Tradescantia pollen tubes.CDPK did not phosphorylate purified rabbit muscle or Dictyostelium actin in vitro. Binding studies showed that CDPK (1) does not cosediment with actin filaments and (2) does not form a complex with G-actin. The data indicate that although CDPK does not interact directly with actin, it may be associated with an actin-binding protein and therefore could play a role in the regulation of the plant cytoskeleton.
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