Intrinsic Tryptophan Fluorescence in the Detection and Analysis of Proteins: A Focus on Förster Resonance Energy Transfer Techniques

International Journal of Molecular Sciences

Amar B. T. Ghisaidoobe²  Sang J. Chung¹ 

[1] Department of Chemistry, Dongguk University, Seoul 100-715, Korea; E-Mail

关键词: FRET;    label free detection;    tryptophan fluorescence;    intrinsic fluorescence;    protein imaging;    biosensors;    immunoassay;   

DOI  :  10.3390/ijms151222518

来源: mdpi

【 摘 要 】

Förster resonance energy transfer (FRET) occurs when the distance between a donor fluorophore and an acceptor is within 10 nm, and its application often necessitates fluorescent labeling of biological targets. However, covalent modification of biomolecules can inadvertently give rise to conformational and/or functional changes. This review describes the application of intrinsic protein fluorescence, predominantly derived from tryptophan (λ_EX ∼ 280 nm, λ_EM ∼ 350 nm), in protein-related research and mainly focuses on label-free FRET techniques. In terms of wavelength and intensity, tryptophan fluorescence is strongly influenced by its (or the protein’s) local environment, which, in addition to fluorescence quenching, has been applied to study protein conformational changes. Intrinsic Förster resonance energy transfer (iFRET), a recently developed technique, utilizes the intrinsic fluorescence of tryptophan in conjunction with target-specific fluorescent probes as FRET donors and acceptors, respectively, for real time detection of native proteins.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.