Dynamic assembly properties of nonmuscle myosin II isoforms revealed by combination of fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy

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概要

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• Myosin II molecules assemble into filaments through their C-terminal rod region, and are responsible for several cellular motile activities. Three isoforms of nonmuscle myosin II (IIA, IIB and IIC) are expressed in mammalian cells. However, little is known regarding the isoform composition in filaments. To obtain new insight into the assembly properties of myosin II isoforms, especially regarding the isoform composition in filaments, we performed a combination analysis of fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS), which enables us to acquire information on both the interaction and the size of each molecule simultaneously. Using C-terminal rod fragments of IIA and IIB (ARF296 and BRF305) labelled with different fluorescent probes, we demonstrated that hetero-assemblies were formed from a mixture of ARF296 and BRF305, and that dynamic exchange of rod fragments occurred between pre-formed homo-assemblies of each isoform in an isoform-independent manner. We also showed that Mts1 (S100A4) specifically stripped ARF296 away from the hetero-assemblies, and consequently, homo-assemblies of BRF305 were formed. These results suggest that IIA and IIB can form hetero-filaments in an isoform-independent manner, and that a factor like Mts1 can remove one isoform from the hetero-filament, resulting in a formation of homo-filaments consisting of another isoform.

• 2011-03-01

著者

• Yazawa Michio

  Division Of Chemistry Graduate School Of Science Hokkaido University

• Sakata Hiroshi

  Laboratory Of Molecular Cell Dynamics Faculty Of Advanced Life Science Hokkaido University

• Sakata Hiroshi

  Laboratory Of Biophysics Ries Hokkaido University

• Takahashi Masayuki

  Division Of Biological Science Graduate School Of Science Hokkaido University

• Mitsuhashi Mariko

  Division Of Chemistry Faculty Of Science Hokkaido University

• KINJO Masataka

  Division of Chemistry, Faculty of Science, Hokkaido University

• Kinjo Masataka

  Division Of Chemistry Faculty Of Science Hokkaido University

• Yazawa Michio

  Division Of Chemistry Faculty Of Science Hokkaido University

• Takahashi Masayuki

  Division Of Chemistry Faculty Of Science Hokkaido University

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