Determination of the affinity of talin and vinculin to charged lipid vesicles: a light scatter study.

Recent experimental findings have demonstrated that both talin and vinculin bind to phospholipids and insert into the hydrophobic region of lipid membranes. Here, we show that the light scatter method can be used for measuring the affinity of proteins to phospholipid membranes. Large unilamellar DMPC/DMPG vesicles were produced by the extrusion technique (LUVETs). We have used repeated heating/cooling scans between 15 degrees C and 35 degrees C to ensure protein-lipid interaction/insertion. A molar affinity of talin, K = 2.9 x 10(6) M-1 and of vinculin, K = 3.3 x 10(5) M-1 to lipid vesicles, respectively, was determined from the plot; light scatter signal at 380 nm against protein concentrations by fitting the term, ln (Io/I-1) = A-K x c to the data.