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

Mitsuhashi, Mariko; Sakata, Hiroshi; Kinjo, Masataka; Yazawa, Michio; Takahashi, Masayuki

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

Mitsuhashi, Mariko; Sakata, Hiroshi; Kinjo, Masataka et al.

The Journal of Biochemistry, 2011, 149(3), 253-263

Permalink : https://hdl.handle.net/2115/47493

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アクセス権	open access
URI	https://hdl.handle.net/2115/47493
タイトル	Dynamic assembly properties of nonmuscle myosin II isoforms revealed by combination of fluorescence correlation spectroscopy and fluorescence cross-correlation spectroscopy
著者	Mitsuhashi, Mariko ; Sakata, Hiroshi ; Kinjo, Masataka ; Yazawa, Michio ; Takahashi, Masayuki
著者名 Mitsuhashi, Mariko
著者名 Sakata, Hiroshi
著者名 Kinjo, Masataka
著者名 Yazawa, Michio
e-Rad_Researcher 50241295 著者名 Takahashi, Masayuki
言語	英語
キーワード	assembly
	FCCS
	myosin II
	FCS
	Mts1 (S100A4)
発行日	2011-03
出版者	Oxford University Press
収録物名	The Journal of Biochemistry
巻(号)	149(3)
ページ	253-263
抄録	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.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	This is a pre-copy-editing, author-produced PDF of an article accepted for publication in The Journal of Biochemistry following peer review. The definitive publisher-authenticated version, J Biochem (2011) 149 (3): 253-263, is available online at: http://jb.oxfordjournals.org/content/149/3/253
PISSN	0021-924X
関連情報	PMID 21106542
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1093/jb/mvq134