title: Immunogold localisation of the intermediate chain within the protein complex of cytoplasmic dynein.
abstract: It was our goal to determine the location of the intermediate chain within the complex of cytoplasmic dynein by immunoelectron microscopy. To do so we generated two monoclonal antibodies (m74-1 and m74-2) specific for the intermediate chain. Both antibodies recognised the intermediate chain by sodium dodecyl sulphate-polyacrylamide gel electrophoresis immunoblot and ELISA assays of native and denatured proteins. When sucrose density gradient-purified cytoplasmic dynein from bovine brain was incubated with the gold-conjugated monoclonal antibodies, m74-1 and m74-2, and examined by negative staining, the gold label was found opposite the globular heads at the base of the V-shaped stalk of the motor complex. The labelling of the intermediate chain is the first mapping of a component within cytoplasmic dynein. The identification of the intermediate chain at the base of the complex supports a possible docking function of the intermediate chain.
authors.affiliation: University of Vienna, Institute of Biochemistry and Molecular Cell Biology, Biocenter, Dr. Bohrgasse 9, A-1030, Vienna, Austria.
authors.forename: W
authors.initials: W
authors.lastname: Steffen
authors.forename: J L
authors.initials: JL
authors.lastname: Hodgkinson
authors.forename: G
authors.initials: G
authors.lastname: Wiche
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D001923:Brain Chemistry; D002417:Cattle; D003593:Cytoplasm / Q000737:chemistry / Q000648:ultrastructure; D004398:Dyneins / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure*; D051379:Mice; D016253:Microscopy, Immunoelectron; D015394:Molecular Structure; D011487:Protein Conformation
chemical_list: D000911:Antibodies, Monoclonal; D004398:Dyneins