Assembly and structure of calcium-induced thick vimentin filaments.
Using a viscometric assay and various electron microscopic procedures (negative staining, rotary shadowing, ultrathin sectioning) we have determined the influences of different kinds of ions and of ionic strength on the structures formed by assembly of soluble subunits of vimentin from bovine lens tissue or from Escherichia coli transformed with Xenopus vimentin cDNA. In contrast to the assembly of typical, i.e., 8 to 14-nm, intermediate-sized filaments (IFs) at elevated (e.g., 160 mM) concentrations of monovalent cations and at millimolar Mg2+ concentrations, filaments formed in the presence of Ca2+ ions (e.g., 5 mM) appeared at a lower rate, attained lower viscosity and were considerably thicker and shorter. The largest diameter measured was that for the recombinant amphibian protein: 24.2 +/- 8.5 nm in negative staining, 28.7 +/- 5.6 nm in sections. These thick Ca(2+)-induced filaments, however, revealed the same approximately 2 nm protofilament composition and approximately 20 nm cross-striation pattern as typical IFs, indicative of a similar molecular arrangement. The significance of this unusual structural IF protein assembly is discussed.
Hofmann
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Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg.
Herrmann
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Franke
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D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002412:Cations / Q000494:pharmacology; D002417:Cattle; D003459:Crystallins / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D007382:Intermediate Filaments / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D016624:Negative Staining; D011994:Recombinant Proteins; D016626:Shadowing Technique, Histology; D014746:Vimentin / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D014783:Viscosity / Q000187:drug effects; D014981:Xenopus
D002412:Cations; D003459:Crystallins; D011994:Recombinant Proteins; D014746:Vimentin; D002118:Calcium