Visualization of monoclonal antibody binding to tropomyosin on native smooth muscle thin filaments by electron microscopy

Fumio Matsumura, Jim Jung Ching Lin

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

We have used three different monoclonal antibodies (LCK16, JLH2 and JLF15) to tropomyosin for the localization of tropomyosin molecules within smooth muscle thin filaments. Thin filaments were incubated with monoclonal antibodies and visualized by negative staining electron microscopy. All three monoclonal antibodies caused the aggregation of thin filaments into ordered bundles, which displayed cross-striations with a periodicity of 37 ± 1 nm. In contrast, conventional rabbit antiserum to tropomyosin distorted and aggregated the thin filaments without generating cross-striations. Therefore, monoclonal antibodies to tropomyosin allow us, for the first time, to observe directly the distribution of tropomyosin molecules along the thin filaments of smooth muscle cells. The binding sites of the antibodies to skeletal muscle tropomyosin were examined by decorating tropomyosin paracrystals with monoclonal antibodies. The LCK16 monoclonal antibody binds the narrow band of tropomyosin paracrystals, whereas the JLF15 antibody binds the wide band of tropomyosin paracrystals.

Original languageEnglish (US)
Pages (from-to)163-171
Number of pages9
JournalJournal of molecular biology
Volume157
Issue number1
DOIs
StatePublished - May 5 1982
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology

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