Nebulin as an actin zipper. A two-module nebulin fragment promotes actin nucleation and stabilizes actin filaments

M. J.G. Chen, C. L. Shih, K. Wang

研究成果: 雜誌貢獻文章同行評審

62 引文 斯高帕斯(Scopus)

摘要

Nebulin is a family of giant muscle proteins (700-900 kDa) that interact with actin to form composite thin filaments in the skeletal muscle sarcomere. This modular protein is composed predominantly of repeating sequence modules of 31-38 residues. To understand the minimum size and number of sequence modules that are required for actin interaction, we studied the behavior of a highly soluble two-module nebulin fragment ND8 that was expressed in Escherichia coli. By fluorescence spectroscopy with pyrenyl-actin and co- sedimentation assays, we observed the following. 1) ND8 greatly accelerated actin nucleation, especially in a buffer that is suboptimal for actin nucleation. The presence of ND8 abolished the lag phase of actin polymerization and increased the net extent of steady state polymerization, thereby reducing the critical concentration of actin polymerization. 2) ND8 reduced the rate of actin depolymerization and might increase the rate of elongation. 3) Cytochalasin E, which caps both ends of actin filaments, inhibited the effect of ND8 on actin polymerization and caused the depolymerization of actin-ND8 complexes. These data suggest that ND8 interacts with actin in such a fashion that it stabilizes the actin nuclei and slows the depolymerization from the ends of actin filaments. 4) The binding stoichiometry of ND8 to F-actin, as estimated by co-sedimentation assays, is 1 to 2 mol of ND8 to 1 mol of actin with an apparent dissociation constant of 20 to 40 μM. Our data suggest that nebulin-actin interaction promotes actin nucleation and stabilizes preformed actin filaments, both of which are desirable attributes of a length-regulating template for actin filaments of the skeletal muscle. Each nebulin molecule may contain as many as 100-200 actin binding domains to form a zipper-like nebulin/actin composite filament.

原文英語
頁(從 - 到)20327-20334
頁數8
期刊Journal of Biological Chemistry
268
發行號27
出版狀態已發佈 - 1993
對外發佈Yes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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