Imaging and nano-manipulation of single biomolecules
著者
和文:
Takashi Funatsu,
Yoshie harada,
Hideo Higuchi,
徳永 万喜洋,
Kiwamu Saito,
Yoshiharu Ishii,
Ronald D. Vale,
Toshio Yanagida.
英文:
Takashi Funatsu,
Yoshie harada,
Hideo Higuchi,
Makio Tokunaga,
Kiwamu Saito,
Yoshiharu Ishii,
Ronald D. Vale,
Toshio Yanagida.
言語
English
掲載誌/書名
和文:
英文:
Biophysical Chemistry
巻, 号, ページ
Vol. 68
pp. 63-72
出版年月
1997年10月
出版者
和文:
英文:
会議名称
和文:
英文:
開催地
和文:
英文:
アブストラクト
We have developed a new technique for imaging single fluorescent dye molecules by refining epifluorescence and totalinternal reflection fluorescence microscopies. In contrast to previously reported single fluorescent molecule imagingmethods, in which specimens were immobilized on an air-dried surface, our method enables video-rate imaging of singlemolecules in aqueous solution. This approach enabled us to directly image the processive movement of individualfluorescently labeled kinesin molecules along a microtubule. This method was also used to visualize individual ATPturnover reactions of single myosin molecules. The method can be combined with molecular manipulation using an opticaltrap. A single kinesin molecule attached to a polystyrene bead was brought into contact with a microtubule adsorbed onto theglass surface. The lifetime of bound Cy%nucleotide in the absence or presence of the microtubule was 10s or 0.08 s,respectively, showing that ATPase activity of the kinesin is strongly activated by microtubules. As the present system isequipped with a nanometer sensor, elemental steps of a single kinesin molecule can also be measured. By simultaneouslymeasuring the individual ATP turnovers and elementary mechanical events of a single kinesin molecule, we will be able toobtain a clear answer to the fundamental problem of how the mechanical events are coupled to the ATPase reaction. 0 1997Elsevier Science B.V.
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