Quantitative analysis of protein-lipid interactions using tryptophan fluorescence.
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.
Kraft
Catherine A
CA
Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Garrido
Jose Luis
JL
Leiva-Vega
Luis
L
Romero
Guillermo
G
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
D008055:Lipids; D008164:Luminescent Proteins; D011506:Proteins; D014364:Tryptophan; D011494:Protein Kinases; C097199:KSR-1 protein kinase; D019908:Proto-Oncogene Proteins c-raf