title: Quantitative analysis of protein-lipid interactions using tryptophan fluorescence.
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.
authors: Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
authors: Catherine A
authors: CA
authors: Kraft
authors: Jose Luis
authors: JL
authors: Garrido
authors: Luis
authors: L
authors: Leiva-Vega
authors: Guillermo
authors: G
authors: Romero
mesh_terms: D008055:Lipids / Q000737:chemistry*; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins; D008722:Methods; D011485:Protein Binding; D011494:Protein Kinases / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D019908:Proto-Oncogene Proteins c-raf / Q000737:chemistry; D014364:Tryptophan / Q000032:analysis* / Q000737:chemistry
chemical_list: D008055:Lipids; D008164:Luminescent Proteins; D011506:Proteins; D014364:Tryptophan; D011494:Protein Kinases; C097199:KSR-1 protein kinase; D019908:Proto-Oncogene Proteins c-raf