title: Polyphosphoinositides inhibit the interaction of vinculin with actin filaments.
abstract: Binding of vinculin to adhesion plaque proteins is restricted by an intramolecular association of vinculin's head and tail regions. Results of previous work suggest that polyphosphoinositides disrupt this interaction and thereby promote binding of vinculin to both talin and actin. However, data presented here show that phosphatidylinositol 4,5-bisphosphate (PI4,5P2) inhibits the interaction of purified tail domain with F-actin. Upon re-examining the effect of PI4,5P2 on the actin and talin-binding activities of intact vinculin, we find that when the experimental design controls for the effect of magnesium on aggregation of PI4,5P2 micelles, polyphosphoinositides promote interactions with the talin-binding domain, but block interactions of the actin-binding domain. In contrast, if vinculin is trapped in an open confirmation by a peptide specific for the talin-binding domain of vinculin, actin binding is allowed. These results demonstrate that activation of the actin-binding activity of vinculin requires steps other than or in addition to the binding of PI4,5P2.
authors.affiliation: Department of Biological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA. pas5@po.cwru.edu
authors.forename: P A
authors.initials: PA
authors.lastname: Steimle
authors.forename: J D
authors.initials: JD
authors.lastname: Hoffert
authors.forename: N B
authors.initials: NB
authors.lastname: Adey
authors.forename: S W
authors.initials: SW
authors.lastname: Craig
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D015153:Blotting, Western; D002645:Chickens; D015636:Magnesium Chloride / Q000494:pharmacology; D008823:Micelles; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000008:administration & dosage / Q000494:pharmacology; D018129:Phosphatidylinositol Phosphates / Q000494:pharmacology*; D016608:Talin / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism*
chemical_list: D000199:Actins; D008823:Micelles; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D016608:Talin; D015636:Magnesium Chloride; D016596:Vinculin