Characterization of single actomyosin rigor bonds: Load dependence of lifetime and mechanical properties

Takayuki Nishizaka, Ryuzo Seo, Hisashi Tadakuma, Kazuhiko Kinosita, Shin'ichi Ishiwata

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Load dependence of the lifetime of the rigor bonds formed between a single myosin molecule (either heavy meromyosin, HMM, or myosin subfragment-1, S1) and actin filament was examined in the absence of nucleotide by pulling the barbed end of the actin filament with optical tweezers. For S1, the relationship between the lifetime (τ) and the externally imposed load (F) at absolute temperature T could be expressed as τ(F) = τ(0)·exp(-F·d/k(B)T) with τ(0) of 67 s and an apparent interaction distance d of 2.4 nm (k(B) is the Boltzmann constant). The relationship for HMM was expressed by the sum of two exponentials, with two sets of τ(0) and d being, respectively, 62 s and 2.7 nm, and 950 s and 1.4 nm. The fast component of HMM coincides with τ(F) for S1, suggesting that the fast component corresponds to single-headed binding and the slow component to double-headed binding. These large interaction distances, which may be a common characteristic of motor proteins, are attributed to the geometry for applying an external load. The pulling experiment has also allowed direct estimation of the number of myosin molecules interacting with an actin filament. Actin filaments tethered to a single HMM molecule underwent extensive rotational Brownian motion, indicating a low torsional stiffness for HMM. From these results, we discuss the characteristics of interaction between actin and myosin, with the focus on the manner of binding of myosin.

Original languageEnglish
Pages (from-to)962-974
Number of pages13
JournalBiophysical Journal
Volume79
Issue number2
Publication statusPublished - 2000
Externally publishedYes

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Actomyosin
Myosins
Actin Cytoskeleton
Myosin Subfragments
Optical Tweezers
Actins
Nucleotides
Temperature
Proteins

ASJC Scopus subject areas

  • Biophysics

Cite this

Nishizaka, T., Seo, R., Tadakuma, H., Kinosita, K., & Ishiwata, S. (2000). Characterization of single actomyosin rigor bonds: Load dependence of lifetime and mechanical properties. Biophysical Journal, 79(2), 962-974.

Characterization of single actomyosin rigor bonds : Load dependence of lifetime and mechanical properties. / Nishizaka, Takayuki; Seo, Ryuzo; Tadakuma, Hisashi; Kinosita, Kazuhiko; Ishiwata, Shin'ichi.

In: Biophysical Journal, Vol. 79, No. 2, 2000, p. 962-974.

Research output: Contribution to journalArticle

Nishizaka, T, Seo, R, Tadakuma, H, Kinosita, K & Ishiwata, S 2000, 'Characterization of single actomyosin rigor bonds: Load dependence of lifetime and mechanical properties', Biophysical Journal, vol. 79, no. 2, pp. 962-974.
Nishizaka, Takayuki ; Seo, Ryuzo ; Tadakuma, Hisashi ; Kinosita, Kazuhiko ; Ishiwata, Shin'ichi. / Characterization of single actomyosin rigor bonds : Load dependence of lifetime and mechanical properties. In: Biophysical Journal. 2000 ; Vol. 79, No. 2. pp. 962-974.
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