Using a gel filtration assay, we have characterized the binding of acidic phospholipids to vinculin. Vinculin binds phosphatidyinositol (Kd ∼ 5 μM) in a reversible, saturable manner in low ionic strength buffers. This interaction is inhibited substantially at 100 mM NaCl and therefore may not be of physiological interest. In contrast, the carboxy-terminal 30-kDa fragment of vinculin, produced by S. aureus V8 protease cleavage, binds acidic phospholipids more tightly than the intact protein, and in a manner insensitive to 100 mM NaCl. Re-addition of the 95-kDa head fragment to the tail restores salt-sensitivity to the tail-lipid interaction. These data indicate that under physiologic ionic conditions, the intramolecular head-tail interaction in vinculin masks a high affinity acidic phospholipid binding site present in the tail domain.
|Original language||English (US)|
|Number of pages||6|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - May 5 1995|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology