Long-Term Observation of Fluorescence of Free Single Molecules To Explore Protein-Folding Energy Landscapes

Autor:	Tamiki Komatsuzaki, Satoshi Takahashi, Yoshitomo Iwahashi, Yoshihiro Sambongi, Kazuya Fujimoto, Kiyoto Kamagata, Toshifumi Kawaguchi, Akinori Baba, Yuji Goto
Rok vydání:	2012
Předmět:	Protein Folding Millisecond Chemistry Confocal Analytical chemistry Curved mirror Energy landscape General Chemistry Biochemistry Fluorescence Catalysis Characterization (materials science) Colloid and Surface Chemistry Protein folding Biological system Energy (signal processing)
Zdroj:	Journal of the American Chemical Society. 134:11525-11532
ISSN:	1520-5126 0002-7863
DOI:	10.1021/ja3020555
Popis:	A method was developed to detect fluorescence intensity signals from single molecules diffusing freely in a capillary cell. A unique optical system based on a spherical mirror was designed to enable quantitative detection of the fluorescence intensity. Furthermore, "flow-and-stop" control of the sample can extend the observation time of single molecules to several seconds, which is more than 1000 times longer than the observation time available using a typical confocal method. We used this method to scrutinize the fluorescence time series of the labeled cytochrome c in the unfolded state. Time series analyses of the trajectories based on local equilibrium state analysis revealed dynamically differing substates on a millisecond time scale. This system presents a new avenue for experimental characterization of the protein-folding energy landscape.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c09a4590d68e1255375bde6145f9d409 https://doi.org/10.1021/ja3020555 Zobrazit plný text záznamu