Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method

Chen Wang; Feng Zhang; Guiying Wang; Ya Cheng; Zhizhan Xu

doi:10.1016/j.ijleo.2008.05.012

Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method

Chen Wang, Feng Zhang, Guiying Wang, Ya Cheng, Zhizhan Xu

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

Fluorescence spectrometry based on fluorescence resonance energy transfer (FRET) principle is a simple but effective tool for investigating protein-protein interactions. In this paper, we report a spectrometry to quantify FRET efficiency based on our home-designed spectral probe system and spectral data-processing procedure. In our method, the fluorescence spectrum from each specimen is recorded at two wavelengths 454 and 502 nm. Least-squares linear fitting algorithm is applied directly to decompose the spectra of donor and acceptor under these two wavelengths to obtain FRET efficiency, which takes both spectral intensity and spectral profile into account compared with traditional three-step analysis. This system and the data-processing procedure enabled us to detect the homo-interaction and hetero-interaction of proteins in living cell.

Original language	English
Pages (from-to)	57-62
Number of pages	6
Journal	Optik
Volume	121
Issue number	1
DOIs	https://doi.org/10.1016/j.ijleo.2008.05.012
State	Published - Jan 2010
Externally published	Yes

Keywords

CFP
FRET
Fluorescence spectroscopy
Spectral unmixing
YFP

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10.1016/j.ijleo.2008.05.012

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title = "Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method",

abstract = "Fluorescence spectrometry based on fluorescence resonance energy transfer (FRET) principle is a simple but effective tool for investigating protein-protein interactions. In this paper, we report a spectrometry to quantify FRET efficiency based on our home-designed spectral probe system and spectral data-processing procedure. In our method, the fluorescence spectrum from each specimen is recorded at two wavelengths 454 and 502 nm. Least-squares linear fitting algorithm is applied directly to decompose the spectra of donor and acceptor under these two wavelengths to obtain FRET efficiency, which takes both spectral intensity and spectral profile into account compared with traditional three-step analysis. This system and the data-processing procedure enabled us to detect the homo-interaction and hetero-interaction of proteins in living cell.",

keywords = "CFP, FRET, Fluorescence spectroscopy, Spectral unmixing, YFP",

author = "Chen Wang and Feng Zhang and Guiying Wang and Ya Cheng and Zhizhan Xu",

year = "2010",

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doi = "10.1016/j.ijleo.2008.05.012",

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volume = "121",

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TY - JOUR

T1 - Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method

AU - Wang, Chen

AU - Zhang, Feng

AU - Wang, Guiying

AU - Cheng, Ya

AU - Xu, Zhizhan

PY - 2010/1

Y1 - 2010/1

N2 - Fluorescence spectrometry based on fluorescence resonance energy transfer (FRET) principle is a simple but effective tool for investigating protein-protein interactions. In this paper, we report a spectrometry to quantify FRET efficiency based on our home-designed spectral probe system and spectral data-processing procedure. In our method, the fluorescence spectrum from each specimen is recorded at two wavelengths 454 and 502 nm. Least-squares linear fitting algorithm is applied directly to decompose the spectra of donor and acceptor under these two wavelengths to obtain FRET efficiency, which takes both spectral intensity and spectral profile into account compared with traditional three-step analysis. This system and the data-processing procedure enabled us to detect the homo-interaction and hetero-interaction of proteins in living cell.

AB - Fluorescence spectrometry based on fluorescence resonance energy transfer (FRET) principle is a simple but effective tool for investigating protein-protein interactions. In this paper, we report a spectrometry to quantify FRET efficiency based on our home-designed spectral probe system and spectral data-processing procedure. In our method, the fluorescence spectrum from each specimen is recorded at two wavelengths 454 and 502 nm. Least-squares linear fitting algorithm is applied directly to decompose the spectra of donor and acceptor under these two wavelengths to obtain FRET efficiency, which takes both spectral intensity and spectral profile into account compared with traditional three-step analysis. This system and the data-processing procedure enabled us to detect the homo-interaction and hetero-interaction of proteins in living cell.

KW - CFP

KW - FRET

KW - Fluorescence spectroscopy

KW - Spectral unmixing

KW - YFP

UR - https://www.scopus.com/pages/publications/70749088297

U2 - 10.1016/j.ijleo.2008.05.012

DO - 10.1016/j.ijleo.2008.05.012

M3 - 文章

AN - SCOPUS:70749088297

SN - 0030-4026

VL - 121

SP - 57

EP - 62

JO - Optik

JF - Optik

IS - 1

ER -

Detection of the homo- and hetero-interaction of proteins using fluorescence resonance energy transfer spectrum method

Abstract

Keywords

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