Real-time imaging of single-molecule fluorescence with a zero-mode waveguide for the analysis of protein-protein interaction

Takeo Miyake, Takashi Tanii, Hironori Sonobe, Rena Akahori, Naonobu Shimamoto, Taro Ueno, Takashi Funatsu, Iwao Ohdomari

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54 Citations (Scopus)

Abstract

Real-time imaging of single-molecule fluorescence with a zero-mode waveguide (ZMW) was achieved. With modification of the ZMW geometry, the signal-to-background ratio is twice that obtainable with a conventional ZMW. The improved signal-to-background ratio makes it possible to visualize individual binding-release events between chaperonin GroEL and cochaperonin GroES at a concentration of 5 μM. Two rate constants representing two-timer kinetics in the release of GroES from GroEL were measured with the ZMW, and the measurements agreed well with those made with a total internal reflection fluorescence microscopy. These results indicate that the novel ZMW makes feasible the direct observation of protein-protein interaction at an intracellular concentration in real time.

Original languageEnglish
Pages (from-to)6018-6022
Number of pages5
JournalAnalytical chemistry
Volume80
Issue number15
DOIs
Publication statusPublished - 2008 Aug 1

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ASJC Scopus subject areas

  • Analytical Chemistry

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