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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 19 (1980), S. 2677-2683 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 25 (1986), S. 435-440 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 14 (1975), S. 3024-3029 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] As a first step towards elucidating the physiological significance of the interaction between PtdInsP2 and cytoskeletal proteins, we stained glycerol-treated chicken skeletal muscle with a monoclonal antibody that is specific for PtdInsP2 (ref. 10). Although PtdInsP2 is a component of the ...
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-2657
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have isolated and purified an activity from amoebae ofPhysarum polycephalum that reduces the flow birefringence of a solution of F-actin in a Ca2+-dependent manner. The purified activity from 100 g of amoebae consisted of 1 mg of a 40000 mol. wt protein. DNase I-affinity chromatography demonstrated that the protein binds toPhysarum actin in a Ca2+-dependent manner, and the binding is not reversed by excess EGTA. Viscometric measurement indicated that the protein (i) accelerates polymerization of G-actin, and (ii) severs F-actin, in a Ca2+-dependent manner. Thus, the protein appeared functionally similar to the fragmin previously isolated fromPhysarum plasmodia (plasmodial fragmin). However, the two proteins had slightly different mobilities on urea-SDS-PAGE, and antibodies raised against the two proteins scarcely cross-reacted with each other. Hence, we conclude that the two proteins are closely related to but are different from each other, and we have named the novel protein ‘myxamoebal fragmin’. Immunoblot analysis indicated that myxamoebal and plasmodial fragmins are specifically present in amoebae and plasmodia, respectively. Results of immunofluorescence staining suggest that the synthesis of plasmodial fragmin is switched on coordinately with the synthesis of the heavy chain of plasmodial myosin and other plasmodium-specific contractile proteins during the apogamic differentiation of amoebae to plasmodia.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 2 (1982), S. 457-470 
    ISSN: 0886-1544
    Keywords: fragmin ; critical actin concentration ; nucleation ; filament growth ; pointed end ; barbed end ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: As reported previously, fragmin isolated form Physarum plasmodia restricts the polymerization of actin to produce short F-actin filaments in the presence of Ca2+ ions. Here it is shown that when actin is polymerized at low concentrations of salts, fragmin increases the critical concentration of actin for polymerization. This effect of fragmin on the critical concentration is independent of the molar ratio of fragmin to actin. The addition of actin monomers onto heavy meromyosin-decorated F-actin fragments treated with fragmin occurs unidirectionally at the pointed end of each fragment. These results suggest that fragmin binds to the barbed ends of F-actin filaments and inhibits association and dissociation of actin monomers at this end. Fragmin accelerates the initial stage of polymerization of actin. When a constant amount of G-actin is polymerized in the presence of small amounts of fragmin, the inverse of the half-polymerization time increases in proportion to the square root of the amount of fragmin added. This means that fragmin acts as a potent promoter of the nucleation step in actin polymerization. Both functions of fragmin-promotion of nucleation and capping at the barbed end of F-actin-require micromolar concentrations of Ca2+.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 0886-1544
    Keywords: cytoplasmic fibril ; birefringence ; microfilament ; contraction-relaxation cycle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The contractility of Physarum plasmodium was investigated using cell models that were prepared by treating thin-spread plasmodia with ice-cold 0.2% Triton X-100. Cell models obtained from the anterior regions of the thin-spread plasmodia in the contraction phase retained many birefringent cytoplasmic fibrils. The fibrils vigorously contracted on addition of ATP, inducing simultaneous contraction of the whole cell models. In contrast, cell models prepared from the anterior regions in the relaxation phase scarcely contained the birefringent fibrils and exhibited only weak contractility on addition of ATP. The posterior regions of the thinspread plasmodia, which were composed of ramified plasmodial strands, always retained many fibrils when treated with the Triton solution and showed intensive contraction on addition of ATP.SDS-polyacrylamide gel electrophoresis showed that the model was enriched for actin and myosin. About 40% of the actin was extracted from the plasmodium by the Triton treatment, while scarcely any myosin was extracted.Fragmin, a F-actin-fragmenting factor, caused the birefringent fibrils to diminish in the presence of Ca2+, but more than 30 minutes was required for their complete disappearance. The birefringent fibrils weakened by 30-minute fragmin treatment disappeared immediately on addition of ATP or AMP-PNP.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 0886-1544
    Keywords: fibroblast ; permeabilized cell model ; Ca2+-dependent contraction ; calmodulin ; phosphorylation ; myosin light chain ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Human lung fibroblast MRC-5 cells treated with Triton X-100 (MRC-5 cell models) were able to contract in the presence of MgATP and Ca2+ of more than 1 μM. Immunofluorescence microscopy with antibodies to actin and myosin 20,000-dalton (20 Kd) light chain revealed that stress fibers were prominent in MRC-5 cell models. Use of a fluorescent actin probe, 7-nitrobenz-2-oxa-1,3-diazole-phallacidin permitted visualization of contraction of the stress fibers in the presence of MgATP and Ca2+. Of the proteins in MRC-5 cell models, only a myosin 20 Kd light chain was phosphorylated in a Ca2+-dependent manner. This Ca2+-dependent phosphorylation of the 20 Kd light chain closely corresponded with the contraction of MRC-5 cell models: 1) Both phosphorylation of the 20 Kd light chain and contraction of MRC-5 cell models were inhibited by calmodulin antagonists such as N-(6-aminohexyl)5-chloro-1-napthalene sulfonamide. 2) The threshold Ca2+ concentration for phosphorylation of the 20 Kd light chain was similar to that for contraction of MRC-5 cell models. Both were lowered by exogenous calmodulin in a concentration-dependent manner. 3) The 20 Kd light chain was thiophosphorylated by incubation of MRC-5 cell models with an ATP analogue, adenosine 5′-0-(3-thiotriphosphate) only in the presence of Ca2+. After this treatment, MRC-5 cell models lost the Ca2+-dependence for contraction. These results indicate that Ca2+-calmodulin-dependent phosphorylation of myosin 20 Kd light chain is required for contraction of MRC-5 cell models.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 10 (1988), S. 410-419 
    ISSN: 0886-1544
    Keywords: calcium ; Ca2+ ; shape change ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When flagellates of Physarum polycephalum were treated with Triton X-100 and more than 10-5 M Ca2+, the microfilamentous cytoskeleton disintegrated, as seen by staining with rhodamine-phalloidin, and myxamoebal fragmin became associated with the Triton-insoluble cytoskeleton as demonstrated by immunofluorescence microscopy and immunoblotting. The association of myxamoebal fragmin with the cytoskeleton was reversed by the subsequent addition of excess EGTA. When flagellates were permeabilized in the absence of Ca2+, myxamoebal fragmin did not associate with the cytoskeleton and diffused out of the cells. Subsequent treatment of these cells with Ca2+ was ineffective in inducing either the association of myxamoebal fragmin with the cytoskeleton or the disintegration of the microfilamentous cytoskeleton. However, treatment of these permeabilized flagellates with 10 μg/ml purified myxamoebal fragmin and 1 mM Ca2+ caused the disintegration of the microfilaments. Therefore, we conclude that myxamoebal fragmin participates in the Ca2+-induced disintegration of the microfilamentous cytoskeleton in these permeabilized cells. Rapid cooling of flagellates caused the reversible association of myxamoebal fragmin with the Triton-insoluble cytoskeleton in vivo. Thus myxamoebal fragmin may also participate in the reorganization of the microfilamentous cytoskeleton induced in vivo by the cold treatment.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 19 (1991), S. 244-254 
    ISSN: 0886-1544
    Keywords: actin regulatory protein ; microtubule-associated protein ; actin bundle ; microtubule bundle ; Physarum ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: An F-actin bundling protein was isolated and purified from plasmodium of Physarum polycephalum. The F-actin bundling protein in Physarum extract was passed through a DEAE-cellulose column. After the protein in the fraction was treated with 6 M urea, it was purified by gel filtration on Sephacryl S-300 HR followed by chromatography on CM-Toyopearl (cation exchange) in the presence of 6 M urea. The purified protein gave a single band on SDS-PAGE, and the molecular weight was estimated to be 52,000. This F-actin bundling protein is referred to as the 52 kDa protein.Interestingly, the 52 kDa protein also induced bundling of microtubules. The formation of F-actin and microtubule bundles was Ca2+-insensitive, but depended on the salt concentration. Each bundle formed at NaCl concentrations less than 0.1 M. The 52 kDa protein cross-reacted with monoclonal antibody raised against a HeLa 55 kDa protein (an F-actin bundling protein from HeLa cells) (Yamashiro-Matsumura and Matsumura: J. Biol. Chem. 260:5087-5097, 1985).When the 52 kDa protein was added to a mixture of actin filaments and microtubules, co-bundles composed of both filaments formed. This is the first reported example in which an F-actin bundling protein induced co-bundling of actin filaments and microtubules.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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