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Sci. Signal., 1 December 2009
Vol. 2, Issue 99, p. pl4
[DOI: 10.1126/scisignal.299pl4]

PROTOCOLS

Quantitative Analysis of Protein-Lipid Interactions Using Tryptophan Fluorescence

Catherine A. Kraft1,*, Jose Luis Garrido1, Luis Leiva-Vega1, and Guillermo Romero1{dagger}

1 The Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
* Present address: Department of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.

Abstract: The fluorescent properties of the amino acid tryptophan make it a useful tool for fluorometric assays. Because tryptophan fluorescence is remarkably sensitive to the polarity of the environment, it can be used to determine the affinity of tryptophan-containing peptides for phospholipid vesicles of varying compositions. Here, we describe a method for using tryptophan fluorescence to determine the binding affinities of peptides derived from the proteins Raf-1 and KSR-1 to small unilamellar vesicles containing phosphatidic acid. The method can be extrapolated to measure the binding of other tryptophan-containing peptides or proteins to lipid vesicles.

{dagger} Corresponding author. E-mail, ggr{at}pitt.edu

Citation: C. A. Kraft, J. L. Garrido, L. Leiva-Vega, G. Romero, Quantitative Analysis of Protein-Lipid Interactions Using Tryptophan Fluorescence. Sci. Signal. 2, pl4 (2009).

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