Mapping proximity within proteins using fluorescence spectroscopy. A study of T4 lysozyme showing that tryptophan residues quench bimane fluorescence

Steven E. Mansoor, Hassane S. Mchaourab, David Farrens

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

We present a novel method for mapping proximity within proteins. The method exploits the quenching of the fluorescent label bimane by nearby Trp residues. In studies of T4 lysozyme we show that this effect appears to be distance dependent and orientation specific. Specifically, we show that a proximal Trp residue can reduce bimane fluorescence intensity by up to 500% and induce complicated fluorescence decay kinetics. Replacing the neighboring Trp residue with phenylalanine removes these spectral perturbations. The advantages of using the Trp quenching of bimane fluorescence for protein structural studies include the low amount of protein required and the substantial simplification of labeling strategies. We anticipate this method will prove suitable for a wide array of high-throughput protein studies such as protein folding, the detection of protein-protein interactions, and, most importantly, the dynamic monitoring of conformational changes.

Original languageEnglish (US)
Pages (from-to)2475-2484
Number of pages10
JournalBiochemistry
Volume41
Issue number8
DOIs
StatePublished - Feb 26 2002

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Fluorescence Spectrometry
Fluorescence spectroscopy
Muramidase
Tryptophan
Fluorescence
Proteins
Quenching
Protein folding
Protein Folding
Phenylalanine
Labeling
bimanes
Labels
Throughput
Kinetics
Monitoring

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mapping proximity within proteins using fluorescence spectroscopy. A study of T4 lysozyme showing that tryptophan residues quench bimane fluorescence. / Mansoor, Steven E.; Mchaourab, Hassane S.; Farrens, David.

In: Biochemistry, Vol. 41, No. 8, 26.02.2002, p. 2475-2484.

Research output: Contribution to journalArticle

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