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Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization

Author

Listed:
  • Joseph N. Forkey

    (University of Pennsylvania)

  • Margot E. Quinlan

    (University of Pennsylvania)

  • M. Alexander Shaw

    (University of Pennsylvania)

  • John E. T. Corrie

    (National Institute for Medical Research)

  • Yale E. Goldman

    (University of Pennsylvania)

Abstract

The structural change that generates force and motion in actomyosin motility has been proposed to be tilting of the myosin light chain domain, which serves as a lever arm. Several experimental approaches have provided support for the lever arm hypothesis; however, the extent and timing of tilting motions are not well defined in the motor protein complex of functioning actomyosin. Here we report three-dimensional measurements of the structural dynamics of the light chain domain of brain myosin V using a single-molecule fluorescence polarization technique that determines the orientation of individual protein domains with 20–40-ms time resolution. Single fluorescent calmodulin light chains tilted back and forth between two well-defined angles as the myosin molecule processively translocated along actin. The results provide evidence for lever arm rotation of the calmodulin-binding domain in myosin V, and support a ‘hand-over-hand’ mechanism for the translocation of double-headed myosin V molecules along actin filaments. The technique is applicable to the study of real-time structural changes in other biological systems.

Suggested Citation

  • Joseph N. Forkey & Margot E. Quinlan & M. Alexander Shaw & John E. T. Corrie & Yale E. Goldman, 2003. "Three-dimensional structural dynamics of myosin V by single-molecule fluorescence polarization," Nature, Nature, vol. 422(6930), pages 399-404, March.
  • Handle: RePEc:nat:nature:v:422:y:2003:i:6930:d:10.1038_nature01529
    DOI: 10.1038/nature01529
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    Cited by:

    1. Ying Xiong & Weijing Han & Chunhua Xu & Jing Shi & Lisha Wang & Taoli Jin & Qi Jia & Ying Lu & Shuxin Hu & Shuo-Xing Dou & Wei Lin & Terence R. Strick & Shuang Wang & Ming Li, 2024. "Single-molecule reconstruction of eukaryotic factor-dependent transcription termination," Nature Communications, Nature, vol. 15(1), pages 1-12, December.
    2. Maes, Christian & O’Kelly de Galway, Winny, 2015. "On the kinetics that moves Myosin V," Physica A: Statistical Mechanics and its Applications, Elsevier, vol. 436(C), pages 678-685.

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