Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

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).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
A simple guide to biochemical approaches for analyzing protein-lipid interactions.
H. Zhao and P. Lappalainen (2012)
Mol. Biol. Cell 23, 2823-2830
   Abstract »    Full Text »    PDF »
Effects of alkyl chain length of gallate on self-association and membrane binding.
E. Takai, A. Hirano, and K. Shiraki (2011)
J. Biochem. 150, 165-171
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882