Immunochemical analysis of the supramolecular structure of myosin in contractile cytoskeletons of Dictyostelium amoebae.
A collection of monoclonal antibodies against Dictyostelium myosin was screened to identify an antibody that could distinguish monomeric from polymeric myosin. An antibody was found that reacted only with monomeric myosin, provided that the antigen-antibody reaction was carried out in solution. This antibody was used in competition radioimmunoassays to probe the supramolecular structure of myosin in Triton-extracted cell models, or cytoskeletons, of Dictyostelium amoebae. The competition assay showed that, as isolated, cytoskeletal myosin was entirely filamentous, but could be converted to monomeric form by increasing the ionic strength of the surrounding buffer. As monomer, it remained associated with the cytoskeleton and could be cycled back to filament form by a second change of buffer. The ability of cytoskeletons to carry out ATP-dependent contraction was examined as a function of the assembly state of myosin. The results suggested that filamentous myosin is responsible for contraction of the cortical filament matrix.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D002021:Buffers; D003599:Cytoskeleton / Q000032:analysis* / Q000502:physiology; D004023:Dictyostelium / Q000648:ultrastructure*; D007163:Immunosorbent Techniques; D046911:Macromolecular Substances; D009218:Myosins / Q000032:analysis / Q000276:immunology / Q000502:physiology; D011863:Radioimmunoassay; D012996:Solutions
D000911:Antibodies, Monoclonal; D002021:Buffers; D046911:Macromolecular Substances; D012996:Solutions; D000255:Adenosine Triphosphate; D009218:Myosins