Imaging of thermal activation of actomyosin motors

Hirokazu Kato, Takayuki Nishizaka, Takashi Iga, Kazuhiko Kinosita, Shin'ichi Ishiwata*

*この研究の対応する著者

研究成果: Article査読

63 被引用数 (Scopus)

抄録

We have developed temperature-pulse microscopy in which the temperature of a microscopic sample is raised reversibly in a square-wave fashion with rise and fall times of several ms, and locally in a region of approximately 10 μm in diameter with a temperature gradient up to 2°C/μm. Temperature distribution was imaged pixel by pixel by image processing of the fluorescence intensity of rhodamine phalloidin attached to (single) actin filaments. With short pulses, actomyosin motors could be activated above physiological temperatures (higher than 60°C at the peak) before thermally induced protein damage began to occur. When a sliding actin filament was heated to 40-45°C, the sliding velocity reached 30 μm/s at 25 mM KCl and 50 μm/s at 50 mM KCl, the highest velocities reported for skeletal myosin in usual in vitro assay systems. Both the sliding velocity and force increased by an order of magnitude when heated from 18°C to 40-45°C. Temperature- pulse microscopy is expected to be useful for studies of biomolecules and cells requiring temporal and/or spatial thermal modulation.

本文言語English
ページ(範囲)9602-9606
ページ数5
ジャーナルProceedings of the National Academy of Sciences of the United States of America
96
17
DOI
出版ステータスPublished - 1999 8月 17
外部発表はい

ASJC Scopus subject areas

  • 遺伝学
  • 一般

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