Use of immobilized lactoperoxidase to label L cell proteins involved in adhesion to polystyrene

Proteins involved in the attachment of murine L cells to polystyrene have been identified by a technique designed to iodinate only those macromolecules coming into closet apposition to the substratum. Whereas soluble lactoperoxidase (LPO) catalyzes the radioiodination of a broad spectrum of polypeptides, the same enzyme immobilized on polystyrene tissue culture flasks discriminately labels 55,000 and 42,000 mol wt polypeptides that adhere tightly to the substratum after the cells are removed. One-dimensional peptide mapping following limited proteolysis showed that the labeled 55,000 mol wt polypeptide is similar to a component of comparable molecular weight present in the detergent- extracted cytoskeleton. The functional association of two cytoskeletal structures, presumably 10-nm filaments and actin, is discussed, and alternative explanations for their susceptibility to iodination by immobilized LPO are presented.

[1]  K. Lanks,et al.  Patterns of proteins synthesized by non-proliferating murine L cells. , 1979, Experimental cell research.

[2]  L. Thornell,et al.  Intermediate (skeletin) filaments in heart Purkinje fibers. A correlative morphological and biochemical identification with evidence of a cytoskeletal function , 1979, The Journal of cell biology.

[3]  E. Lazarides,et al.  Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments , 1979, The Journal of cell biology.

[4]  B. L. Granger,et al.  The existence of an insoluble Z disc scaffold in chicken skeletal muscle , 1978, Cell.

[5]  E. Wang,et al.  Functions of cytoplasmic fibers in intracellular movements in BHK-21 cells , 1978, The Journal of cell biology.

[6]  Kenneth M. Yamada,et al.  Fibronectins—adhesive glycoproteins of cell surface and blood , 1978, Nature.

[7]  K. Yamada,et al.  Immunological characterization of a major transformation-sensitive fibroblast cell surface glycoprotein. Localization, redistribution, and role in cell shape , 1978, The Journal of cell biology.

[8]  D. Lukeman,et al.  Actin content and organization in normal and transformed cells in culture , 1978, The Journal of cell biology.

[9]  R. Goldman,et al.  Biochemical and immunological analysis of rapidly purified 10-nm filaments from baby hamster kidney (BHK-21) cells , 1978, The Journal of cell biology.

[10]  F. Grinnell.  Cellular adhesiveness and extracellular substrata. , 1978, International review of cytology.

[11]  A. Woods,et al.  Mechanisms of cellular adhesion. II. The interplay between adhesion, the cytoskeleton and morphology in substrate-attached cells. , 1977, Experimental Cell Research.

[12]  K. Lanks,et al.  Covalent attachment of lactoperoxidase to polystyrene tissue culture flasks. , 1977, Analytical biochemistry.

[13]  R. Hynes,et al.  Surface distribution of LETS protein in relation to the cytoskeleton of normal and transformed cells , 1977, The Journal of cell biology.

[14]  R A Laskey,et al.  Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film , 1977, FEBS letters.

[15]  L. Culp,et al.  Morphology and cellular origins of substrate-attached material from mouse fibroblasts. , 1977, Experimental cell research.

[16]  R. Hynes,et al.  Restoration of normal morphology, adhesion and cytoskeleton in transformed cells by addition of a transformation-sensitive surface protein , 1977, Cell.

[17]  M. Kirschner,et al.  Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. , 1977, The Journal of biological chemistry.

[18]  T. Chen,et al.  In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. , 1977, Experimental cell research.

[19]  E. Lazarides,et al.  Immunological characterization of the subunit of the 100 A filaments from muscle cells. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[20]  L. Culp.  Electrophoretic analysis of substrate-attached proteins from normal and virus-transformed cells. , 1976, Biochemistry.

[21]  G. Nicolson.  Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over surface components. , 1976, Biochimica et biophysica acta.

[22]  I. Pastan,et al.  Cell surface protein partially restores morphology, adhesiveness, and contact inhibition of movement to transformed fibroblasts. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[23]  J. Brown,et al.  Identification of a high molecular weight trans-membrane protein in mouse L cells. , 1975, Progress in clinical and biological research.

[24]  J. Spudich,et al.  Cytoskeletal elements of chick embryo fibroblasts revealed by detergent extraction. , 1976, Journal of supramolecular structure.

[25]  L. Culp,et al.  Molecular composition and origin of substrate-attached material from normal and virus-transformed cells. , 1976, Journal of supramolecular structure.

[26]  S. Brecher.  The occurrence and possible role of 80-100 A filaments in PtKl cells. , 1975, Experimental cell research.

[27]  R. Goldman,et al.  The use of heavy meromyosin binding as an ultrastructural cytochemical method for localizing and determining the possible functions of actin-like microfilaments in nonmuscle cells. , 1975, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[28]  J. Spudich,et al.  Visualization of actin fibers associated with the cell membrane in amoebae of Dictyostelium discoideum. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[29]  S. Granett,et al.  Ultrastructural localization of the high molecular weight proteins associated with in vitro-assembled brain microtubules , 1975, The Journal of cell biology.

[30]  K. Weber,et al.  Patterns of organization of actin and myosin in normal and transformed cultured cells. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[31]  A. Rich,et al.  Decrease in membrane-associated actin of fibroblasts after transformation by Rous sarcoma virus. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[32]  A. Dipasquale,et al.  THE UPPER CELL SURFACE: ITS INABILITY TO SUPPORT ACTIVE CELL MOVEMENT IN CULTURE , 1974, The Journal of cell biology.

[33]  E. Lazarides,et al.  Actin antibody: the specific visualization of actin filaments in non-muscle cells. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[34]  D. Ho,et al.  Adhesion of culture cells to their substratum. , 1974, Experimental cell research.

[35]  S. D. Petris,et al.  THE LYMPHOCYTE PLASMA MEMBRANE , 1974 .

[36]  B. Zimmerman.  Lymphocyte plasma membranes. I. Characterization of an antigenic plasma membrane constituent on rat thymus cells. , 1974, Journal of immunology.

[37]  A. Harris.  Location of cellular adhesions to solid substrata. , 1973, Developmental biology.

[38]  J. Uhr,et al.  Isolation and characterization of H-2 and TL alloantigens from the surface of mouse lymphocytes. , 1972, Cellular immunology.

[39]  J. Pontén,et al.  Specialization of cell surfaces in contact-inhibited human glia-like cells in vitro. , 1971, Experimental cell research.

[40]  C. Heidelberger,et al.  Quantitative studies on the malignant transformation of mouse prostate cells by carcinogenic hydrocarbons in vitro , 1969, International journal of cancer.

[41]  I. Macpherson,et al.  Studies on transformation of hamster cells by polyoma virus in vitro , 1961 .

[42]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[43]  D. R. Coman.  Decreased Mutual Adhesiveness, a Property of Cells from Squamous Cell Carcinomas , 1944 .