Fibronectin and the cytoskeleton of epithelial cells

Tolson, N.D.; Hopkins, Colin R.; Sheterline, Peter; Schor, Seth L.

doi:10.1016/0309-1651(80)90118-6

Cited by 3 publications

(6 citation statements)

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“…It has been established elsewhere (see reference 19), and confirmed in this laboratory (38), that treatment of cells attached to culture dishes with nonionic detergents under controlled conditions removes the majority ofmembrane-bounded organelles and soluble proteins. The detergent-insoluble material remaining on the dish consists mainly of microfilaments, microtubules, and 10-nm filaments, which retain a distribution similar to that seen in fixed and permealysed cells.…”

Section: Evaluation Of the Lysis Proceduressupporting

confidence: 54%

“…Note that the band with an apparent molecular weight of 217,000, which migrates between myosin and actin-binding protein, is similar in size to fibronectin . We have recently shown an association between fibronectin and stress fibers on the dorsal surface of detergent-insoluble ghosts prepared from ovarian granulosa cells (38). DISCUSSION The experiments described above were initiated to find a means of studying the molecular events occurring at the cell surface after binding and redistribution of lectin.…”

Section: Effects Of Lectin On Neutrophil Cell Surface Motilitymentioning

confidence: 99%

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Transmembrane linkage between surface glycoproteins and components of the cytoplasm in neutrophil leukocytes.

Sheterline¹,

Hopkins²

1981

The Journal of Cell Biology

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An experimental approach is described that enables the analysis of interactions between exogenous surface ligands and components of the cytoplasm in neutrophil leukocytes . Neutrophils treated with the nonionic detergent Lubrol FIX, under controlled conditions, yield intact detergent-insoluble ghosts . Morphological analysis of neutrophil ghosts shows that they retain the original dimensions of the cell and consist almost entirely of a peripheral filamentous network, representing the submembranous cortical web, concentric to nuclear remnants . All intracellular membrane-bounded organelles, plasma membrane, and background cytoplasmic electron density are absent .Biochemical analysis of the ghosts shows that <10% of enzyme markers for the soluble and granule fractions remain, and that >90% of total cell phospholipid is removed during detergent extraction . The major proteins remaining in the ghosts comigrate, on polyacrylamide gels in the presence of SDS, with chicken gizzard actin, myosin, filamin, and a 110-kdalton protein .Patches and caps induced on neutrophils with either fluorescein isothiocyanate-concanavalin A or ferritin-concanavalin A retain their original location and morphology on ghosts after lysis, as determined by both fluorescence and electron microscopy . In similar experiments, but using '25 1-labeled lectins, 37% of total cell bound concanavalin A (Con A) and 25% succinylated Con A remain attached to the ghosts .A major "51-labeled membrane glycoprotein (80 kdaltons) is associated with ghosts prepared from intact neutrophils iodinated in the presence of exogenous lactoperoxidase . Further ' 25 1-labeled membrane glycoproteins (217, 170, and 147 kdaltons) become associated with ghosts prepared from iodinated cells treated before lysis with Con A, but not with succinylated Con A.These data taken together suggest that linkages exist in neutrophils between proteins exposed on the outer surface of the plasma membrane and the peripheral filamentous network independent of the presence of lipid bilayer. The implications of these findings for surface motile phenomena will be discussed .Since the wide acceptance of a membrane structure based on the fluid mosaic model proposed by Singer and Nicolson (35), it has become apparent that the lateral mobility of some membrane proteins may be constrained or influenced by their interaction, through the lipid bilayer, with components of the cytoplasm (14). This property of certain membrane proteins can be inferred from their behavior in the presence of multivalent ligands (34) and from lateral diffusion data obtained in photobleaching experiments (30,31,36) . In some cases these phenomena can be enhanced or inhibited by treatment of cells with agents that interfere with microtubule or microfilament THE JOURNAL OF CELL BIOLOGY " VOLUME 90 SEPTEMBER 1981 743-754 © The Rockefeller University Press " 0021-9525/81/09/0743/12 $1 .00 integrity (13,14), thus leading to the predominant view that these structures are somehow involved either directly or indirectly in...

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Section: Evaluation Of the Lysis Proceduressupporting

confidence: 54%

Section: Effects Of Lectin On Neutrophil Cell Surface Motilitymentioning

confidence: 99%

Transmembrane linkage between surface glycoproteins and components of the cytoplasm in neutrophil leukocytes.

Sheterline¹,

Hopkins²

1981

The Journal of Cell Biology

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“…70 nm apart) the density of Concanavalin A binding sites of neutrophilic leucocytes is -1 x 1061 cell (i.e. 5-10 nm apart) (Sheterline, 1980). In non-random distributions, as for example where receptors may occur within small clusters (McKanna et al, 1979) or within coated vesicles (-60-80 nm dia.…”

Section: Discussionmentioning

confidence: 99%

“…For these studies rabbit antisera were raised to porcine plasma fibronectin (Tolson et al, 1980). A conventional indirect immunofluorescence method using fluorescine labelled goat anti-rabbit antiserum displayed the distribution of fibronectin on the upper surface of the cultured cells and essentially the same localization in TEM was achieved using antifibronectin followed by gold/protein A.…”

Section: ) Localization Of Jibronectin On Ovarian Granulosa Cellsmentioning

confidence: 99%

“…9, networks of fibronectin are identified by the distribution of gold particles on the upper cell surface. Fibronectin beneath intact cells is inaccessible to antibody (Hynes et al, 1976;Tolson et al, 1980), but where cells have peeled away from the substratum clearly labelled 'footprints' are seen (Fig. 10).…”

Section: ) Localization Of Jibronectin On Ovarian Granulosa Cellsmentioning

confidence: 99%

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Cell surface labelling with gold colloid particulates: the use of avidin and staphylococcal protein A‐coated gold in conjunction with biotin and fc‐bearing ligands

Tolson

Boothroyd

Hopkins

1981

Journal of Microscopy

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SUMMARY Procedures for preparing gold colloid particles stabilized with either avidin or protein A are described. Methods of using these general utility tracers for localizing biotinylated and fc bearing immunoglobulins are outlined and, as examples of the way in which these methods can be applied, procedures for identifying epidermal growth factor receptors and surface fibronectin on ovarian granulosa cells are described.

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Factors Affecting Mammary Cells in Culture

Kano-Sueoka¹

1983

Biochemical Actions of Hormones

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Fibronectin and the cytoskeleton of epithelial cells

Cited by 3 publications

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Transmembrane linkage between surface glycoproteins and components of the cytoplasm in neutrophil leukocytes.

Transmembrane linkage between surface glycoproteins and components of the cytoplasm in neutrophil leukocytes.

Cell surface labelling with gold colloid particulates: the use of avidin and staphylococcal protein A‐coated gold in conjunction with biotin and fc‐bearing ligands

Factors Affecting Mammary Cells in Culture

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