Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Analysis of cytokeratin domains by cloning and expression of intact and deleted polypeptides in Escherichia coli.
Magin TM, Hatzfeld M, Franke WW.
???displayArticle.abstract???
Using recombination of an appropriate expression vector system (pINDU) with a complete cDNA encoding a basic (type II) cytokeratin, i.e. cytokeratin 8 (1) of Xenopus laevis, we transformed Escherichia coli cells to synthesize considerable amounts of an insoluble eukaryotic cytoskeletal protein. The cytokeratin was deposited in large 'inclusion bodies' in the bacterial cytoplasm but did not form detectable filamentous structures. However, when the E. coli-expressed cytokeratin was purified and combined in vitro with an authentic cytokeratin of the complementary, i.e. acidic (type I) subfamily, it formed typical intermediate-sized filaments (IFs). Using Bal31 deletion from either the 5' or the 3' end of the cDNA, series of polypeptides progressively deleted from the amino or the carboxy terminus were produced in E. coli and identified by monoclonal antibodies. These assays allowed the mapping of epitopes. The deletion polypeptides of cytokeratin 8 were further examined to localize the region(s) involved in the heterotypic binding of alpha-helices of type I cytokeratins, using an in vitro nitrocellulose blot binding assay. We show that a region of 37 amino acids located in the central portion of coil 2 of the alpha-helical rod domain is sufficient for the specific recognition of a radiolabelled type I cytokeratin, i.e. cytokeratin 18 (D) from rat liver. In addition, deletion polypeptides containing only coil 1 of the alpha-helical rod also bind strongly the complementary cytokeratin. This indicates that the capability of heterotypic recognition and complex formation is not restricted to a single signal sequence but is located in distant and independent alpha-helical domains.(ABSTRACT TRUNCATED AT 250 WORDS)
Achtstaetter,
Separation of cytokeratin polypeptides by gel electrophoretic and chromatographic techniques and their identification by immunoblotting.
1986, Pubmed
Achtstaetter,
Separation of cytokeratin polypeptides by gel electrophoretic and chromatographic techniques and their identification by immunoblotting.
1986,
Pubmed Aebi,
The fibrillar substructure of keratin filaments unraveled.
1983,
Pubmed Bader,
Amino acid sequence and gene organization of cytokeratin no. 19, an exceptional tail-less intermediate filament protein.
1986,
Pubmed Cheng,
Increased cell buoyant densities of protein overproducing Escherichia coli cells.
1983,
Pubmed Cowin,
Plakoglobin: a protein common to different kinds of intercellular adhering junctions.
1986,
Pubmed Eichner,
The role of keratin subfamilies and keratin pairs in the formation of human epidermal intermediate filaments.
1986,
Pubmed Franke,
Identification and characterization of epithelial cells in mammalian tissues by immunofluorescence microscopy using antibodies to prekeratin.
1979,
Pubmed Franke,
Protein complexes of intermediate-sized filaments: melting of cytokeratin complexes in urea reveals different polypeptide separation characteristics.
1983,
Pubmed Franke,
Identification of the conserved, conformation-dependent cytokeratin epitope recognized by monoclonal antibody (lu-5).
1987,
Pubmed
,
Xenbase Franke,
Diversity of cytokeratins. Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues.
1981,
Pubmed Franke,
HeLa cells contain intermediate-sized filaments of the prekeratin type.
1979,
Pubmed Franke,
Different intermediate-sized filaments distinguished by immunofluorescence microscopy.
1978,
Pubmed Franz,
Cloning of cDNA and amino acid sequence of a cytokeratin expressed in oocytes of Xenopus laevis.
1986,
Pubmed
,
Xenbase Fuchs,
Two distinct classes of keratin genes and their evolutionary significance.
1981,
Pubmed Geisler,
Antiparallel orientation of the two double-stranded coiled-coils in the tetrameric protofilament unit of intermediate filaments.
1985,
Pubmed Geisler,
The amino acid sequence of chicken muscle desmin provides a common structural model for intermediate filament proteins.
1982,
Pubmed Geisler,
Neurofilament architecture combines structural principles of intermediate filaments with carboxy-terminal extensions increasing in size between triplet proteins.
1983,
Pubmed Geisler,
Proteinchemical characterization of three structurally distinct domains along the protofilament unit of desmin 10 nm filaments.
1982,
Pubmed Gruen,
Structural studies on the microfibrillar proteins of wool. Interaction between alpha-helical segments and reassembly of a four-chain structure.
1983,
Pubmed Hanukoglu,
The cDNA sequence of a Type II cytoskeletal keratin reveals constant and variable structural domains among keratins.
1983,
Pubmed Hanukoglu,
The cDNA sequence of a human epidermal keratin: divergence of sequence but conservation of structure among intermediate filament proteins.
1982,
Pubmed Hatzfeld,
Pair formation and promiscuity of cytokeratins: formation in vitro of heterotypic complexes and intermediate-sized filaments by homologous and heterologous recombinations of purified polypeptides.
1985,
Pubmed Heid,
The complement of native alpha-keratin polypeptides of hair-forming cells: a subset of eight polypeptides that differ from epithelial cytokeratins.
1986,
Pubmed Ip,
Assembly of vimentin in vitro and its implications concerning the structure of intermediate filaments.
1985,
Pubmed Kaufmann,
Intermediate filament forming ability of desmin derivatives lacking either the amino-terminal 67 or the carboxy-terminal 27 residues.
1985,
Pubmed Kyhse-Andersen,
Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose.
1984,
Pubmed Lebel,
Lamins A, B and C share an epitope with the common domain of intermediate filament proteins.
1987,
Pubmed Lee,
Organisation of the polypeptide chains in mammalian keratin.
1976,
Pubmed Magin,
Translational products of mRNAs coding for non-epidermal cytokeratins.
1983,
Pubmed
,
Xenbase Markwell,
A new solid-state reagent to iodinate proteins. I. Conditions for the efficient labeling of antiserum.
1982,
Pubmed Maxam,
Sequencing end-labeled DNA with base-specific chemical cleavages.
1980,
Pubmed Moll,
The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells.
1982,
Pubmed Osborn,
Cytoplasmic intermediate filament proteins and the nuclear lamins A, B and C share the IFA epitope.
1987,
Pubmed Parry,
The coiled-coil molecules of intermediate filaments consist of two parallel chains in exact axial register.
1985,
Pubmed Paul,
Immunocytochemical demonstration of human proinsulin chimeric polypeptide within cytoplasmic inclusion bodies of Escherichia coli.
1983,
Pubmed Pruss,
All classes of intermediate filaments share a common antigenic determinant defined by a monoclonal antibody.
1981,
Pubmed Quax,
The structure of the vimentin gene.
1983,
Pubmed Quinlan,
Characterization of dimer subunits of intermediate filament proteins.
1986,
Pubmed Quinlan,
Heterotypic tetramer (A2D2) complexes of non-epidermal keratins isolated from cytoskeletons of rat hepatocytes and hepatoma cells.
1984,
Pubmed Quinlan,
Patterns of expression and organization of cytokeratin intermediate filaments.
1985,
Pubmed Renner,
Reconstitution of intermediate-sized filaments from denatured monomeric vimentin.
1981,
Pubmed Schiller,
A subfamily of relatively large and basic cytokeratin polypeptides as defined by peptide mapping is represented by one or several polypeptides in epithelial cells.
1982,
Pubmed Steinert,
The molecular biology of intermediate filaments.
1985,
Pubmed Steinert,
Ten-nanometer filaments of hamster BHK-21 cells and epidermal keratin filaments have similar structures.
1978,
Pubmed Steinert,
Intermediate filaments of baby hamster kidney (BHK-21) cells and bovine epidermal keratinocytes have similar ultrastructures and subunit domain structures.
1980,
Pubmed Steinert,
Self-assembly of bovine epidermal keratin filaments in vitro.
1976,
Pubmed Stueber,
A novel in vitro transcription-translation system: accurate and efficient synthesis of single proteins from cloned DNA sequences.
1984,
Pubmed Sun,
Keratin cytoskeletons in epithelial cells of internal organs.
1979,
Pubmed Sun,
Immunofluorescent staining of keratin fibers in cultured cells.
1978,
Pubmed Thomas,
An octamer of histones in chromatin and free in solution.
1975,
Pubmed Traub,
Involvement of the N-terminal polypeptide of vimentin in the formation of intermediate filaments.
1983,
Pubmed Tseng,
Correlation of specific keratins with different types of epithelial differentiation: monoclonal antibody studies.
1982,
Pubmed Ward,
Cross-linking of Novikoff ascites hepatoma cytokeratin filaments.
1985,
Pubmed Weber,
Intermediate filaments: structural conservation and divergence.
1985,
Pubmed Williams,
Cytoplasmic inclusion bodies in Escherichia coli producing biosynthetic human insulin proteins.
1982,
Pubmed Woods,
The number of polypeptide chains in the rod domain of bovine epidermal keratin.
1983,
Pubmed
