Electron transfer reaction in a single protein molecule observed by total internal reflection fluorescence microscopy

Yoshiaki Furukawa, Tadato Ban, Daizo Hamada, Koichiro Ishimori, Yuji Goto, Isao Morishima

研究成果: Article査読

11 被引用数 (Scopus)

抄録

To observe an electron transfer (ET) process in a single protein molecule, we constructed a model system, Alexa-HCytb5, in which cytochrome b5 (Cytb5) is modified with a fluorescent probe, Alexa Fluor 647 dye. In this model system, intramolecular transfer of an electron from the Alexa dye to heme in Cytb5 is supposed to oxidize the probe and quench its fluorescence, and the ET reaction at the single-molecule level can be monitored as the intermittent change in the fluorescence intensity. Alexa-HCytb5 was fixed on the glass surface, and illumination of laser light by the total internal reflection resulted in blinking of the fluorescence from the single Alexa-HCytb5 molecule in the time scale of several hundred milliseconds. Each Alexa-HCytb5 molecule is characterized by its own rate constant of the blinking, corresponding to the ET rate constant at the single-molecule level, and its variation ranges between 1 and 10 s-1. The current system thus enables us to visualize the ET reaction in the single protein molecule, and the protein ET reaction was found to be explained by the distribution of the rate constants. On the basis of the Marcus theory, we suggest that the origin of this rate distribution is the distance change associated with the structural fluctuation in the protein molecule.

本文言語English
ページ(範囲)2098-2103
ページ数6
ジャーナルJournal of the American Chemical Society
127
7
DOI
出版ステータスPublished - 2005 2月 23
外部発表はい

ASJC Scopus subject areas

  • 触媒
  • 化学一般
  • 生化学
  • コロイド化学および表面化学

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