Search PubMed⌕ Search

Biomedical subjects

K Burridge

Publications and source records attributed to K Burridge.

121 records · Page 7Linked to original sources
PubMed1978

Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum.

An antiserum has been found in a nonimmunized rabbit which reacts strongly with a system of filaments in various fibroblasts, epithelial cells, macrophages and neuroblastoma. These filaments are distinct from the actin microfilament bundles visualized by an antibody against actin, and they are not affected by brief treatment with cytochalasin B. The pattern of these filaments somewhat resembles that described for microtubules, but the filaments could be clearly distinguished from microtubules by a comparison of their respective immunofluorescent patterns during cell division. In response to the drugs colcemid and vinblastine, the filaments reacting with this preimmune serum condense to form a compact perinuclear coil of fibers, a distribution and behavior in agreement with that previously described for the 10 nm or intermediate filaments studied by electron microscopy. Further evidence supporting our conclusion that this antiserum reacts with intermediate filaments is provided by a comparison of electron micrographs and the immunofluorescent patterns from parallel cell cultures. To identify the antigens reacting with this antiserum we have used the new technique of immuno-autoradiography on SDS gels of whole cell extracts. Two reactive polypeptide chains have been identified with apparent molecular weights of 56,000 and 30,000 daltons.

Animals↗
PubMed1976

Changes in cellular glycoproteins after transformation: identification of specific glycoproteins and antigens in sodium dodecyl sulfate gels.

The glycoproteins of whole cells have been analyzed by direct application of radio-iodinated lectins to sodium dodecyl sulfate gels, followed by autoradiography. By use of lectins with different carbohydrate specificities, different sets of glycoproteins have been visualized. The most prominent lectin-binding band in many gels is thelarge, external, transformation-sensitive (LETS) protein. Major glycoprotein differences are revealed when normal and virus-transformed cells are compared. Certain differences, however, are also seen when the glycoproteins are compared from two separately derived simian virus 40 transformants of 3T3 cells, suggesting a degree of clonal variation between these lines that may not relate to transformation. A complementary technique is used to detect specific antigens in sodium dodecyl sulfate gels by direct application of iodinated antibodies. An antiserum specific for the LETS protein is used to identify this antigen in the gels of both normal and transformed cells.

Antigens, Neoplasm↗
PubMed1975

Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells.

Antibodies specific for the skeletal muscle structural protein alpha-actinin are used to localize this protein by indirect immunofluorescence in nonmuscle cells. In cultured nonmuscle cells, alpha-actinin is localized along or between actin filament bundles producing an almost regular periodicity. The protein is also detected in the form of fluorescent plaques at some ends of actin filament bundles, as well as in a filamentous form in some overlap areas of cells. In spreading rat embryo cells, alpha-actinin assumes a focal distribution which corresponds to the vertices of a highly regular actin filament network. The results suggest that alpha-actinin may be involved in the organization of actin filament bundles, in the attachment of actin filaments to the plasma membrane, and in the assembly of actin filaments in areas of cell to cell contact.

Actinin↗
PubMedDate not supplied

An interaction between vinculin and talin.

In cultured fibroblasts, microfilament bundles terminate at adhesion plaques (focal contacts), the specialized regions where the cells adhere most tightly to the underlying substrate. Vinculin is a protein concentrated in adhesion plaques and has been suggested as a possible link between the ends of the bundles of actin filaments and the plasma membrane. If vinculin is one protein in a chain of attachment between the bundles of microfilaments and the plasma membrane, it is important to identify other components which interact with vinculin. We have recently discovered a new protein in adhesion plaques which we refer to as talin. Here we show that talin binds to vinculin, which suggests that talin may be involved with vinculin in the attachment of microfilament bundles to the plasma membrane at the adhesion plaques.

Animals↗
PubMedDate not supplied

Identification of talin as a major cytoplasmic protein implicated in platelet activation.

During platelet activation there is a major reorganization in the platelet cytoskeleton that accompanies a rapid change in platelet shape. Many of the events associated with activation are attributed to a rise in calcium concentration within the platelet cytoplasm. One direct consequence of the elevated calcium is the activation of a calcium-dependent protease that cleaves a major platelet protein of relative molecular mass (Mr) approximately 235,000 (235K) to 200K. This protein, P235, has been purified and reported to interact with actin, but the significance of the proteolytic cleavage is unknown. Talin, a cytoskeletal protein in smooth muscle and fibroblasts, binds vinculin and, together with vinculin, is localized in fibroblasts at sites of actin-membrane attachment. Talin and P235 have similar purification procedures, sedimentation coefficients and Stokes' radii (ref. 6 and Molony et al., unpublished observations). Of particular significance, talin is readily cleaved by proteases from approximately 215K to a fragment of approximately 190K. Given these similarities we have investigated the possible relationship between these proteins. Here we demonstrate that platelet P235 is recognized by anti-talin antibody and that it binds vinculin. Both proteins are cleaved in vitro by the calcium-activated protease to yield similar fragments. We conclude that P235 corresponds to the platelet form of talin.

Actins↗
PubMedDate not supplied

Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage.

Many observations suggest the presence of transmembrane linkages between the cytoskeleton and the extracellular matrix. In fibroblasts both light and electron microscopic observations reveal a co-alignment between actin filaments at the cell surface and extracellular fibronectin. These associations are seen at sites of cell matrix interaction, frequently along stress fibres and sometimes where these bundles of microfilaments terminate at adhesion plaques (focal contacts). Non-morphological evidence also indicates a functional linkage between the cytoskeleton and extracellular matrix. Addition of fibronectin to transformed cells induces flattening of the cells and a reorganization of the actin cytoskeleton, with the concomitant appearance of arrays of stress fibres. Conversely, disruption of the actin cytoskeleton by treatment with cytochalasin B leads to release of fibronectin from the cell surface. As yet, there is no detailed knowledge of the molecules involved in this transmembrane linkage, although several proteins have been suggested as candidates in the chain of attachment between bundles of actin filaments and the cytoplasmic face of the plasma membrane: these include vinculin, alpha-actinin and talin, each one having been identified at regions where bundles of actin filaments interact with the plasma membrane and underlying cell-surface fibronectin. Recently, the cell-substrate attachment (CSAT) antigen has been identified as a plasma membrane receptor for fibronectin, raising the possibility that this glycoprotein complex may serve as a bridge between fibronectin and one or more of the underlying cytoskeletal components mentioned. Here we have investigated the interaction of the purified CSAT antigen with these cytoskeletal components, and we demonstrate an interaction specifically between the CSAT antigen and talin.

Animals↗