Tropomyosin-enriched and alpha-actinin-enriched microfilaments isolated from chicken embryo fibroblasts by monoclonal antibodies

J. J.C. Lin, F. Matsumura, S. Yamashiro Matsumura

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Abstract

Antitropomyosin and anti-alpha-actinin monoclonal antibodies have been used to isolate two classes of microfilaments, i.e., tropomyosin-enriched and alpha-actinin-enriched microfilaments, respectively, from cultured chicken embryo fibroblasts. Electron microscopic studies of the isolated tropomyosin-enriched microfilaments showed periodic localization of tropomyosin along the microfilaments, with a 35-nm repeat. On the contrary, the isolated alpha-actinin-enriched microfilaments showed no obvious periodicity. Many individual alpha-actinin-enriched microfilaments with length >1 μm (ranging from 1 to 10 μm) were aggregated by anti-alpha-actinin monoclonal antibodies. Both of the isolatd microfilaments had the ability to activate the Mg2+-ATPase activity of skeletal muscle myosin, although different extents of activation were observed. These two classes of microfilaments also differed in their protein composition. Molar ratios of major identifiable proteins in the isolated microfilaments were alpha-actinin(dimer):actin(monomer):tropomyosin(dimer) = <0.02:8.06:1.00 for tropomyosin-enriched microfilaments and 0.44:13.91:1.00 for alpha-actinin-enriched microfilaments. By two-dimensional gel analysis of the isolated microfilaments, we have found seven spots which possess typical tropomyosin properties including pl 4.5, immunological cross-reaction, lack of proline and tryptophan, and heat stability. Pulse-chase experiments suggested that the assembly of microfilament-associated proteins, at least for alpha-actinin and tropomyosins, was coordinately regulated by the assembly of actin into microfilaments.

Original languageEnglish (US)
Pages (from-to)116-127
Number of pages12
JournalJournal of Cell Biology
Volume98
Issue number1
DOIs
StatePublished - 1984
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Cell Biology

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