title: Distinct specificity in the recognition of phosphoinositides by the pleckstrin homology domains of dynamin and Bruton's tyrosine kinase.
abstract: Pleckstrin homology (PH) domains may act as membrane localization modules through specific interactions with phosphoinositide phospholipids. These interactions could represent responses to second messengers, with scope for regulation by soluble inositol polyphosphates. A biosensor-based assay was used here to probe interactions between PH domains and unilamellar liposomes containing different phospholipids and to demonstrate specificity for distinct phosphoinositides. The dynamin PH domain specifically interacted with liposomes containing phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] and, more weakly, with liposomes containing phosphatidylinositol-4-phosphate [PI(4)P]. This correlates with phosphoinositide activation of the dynamin GTPase. The functional GTPase of a dynamin mutant lacking the PH domain, however, cannot be activated by PI(4,5)P2. The phosphoinositide-PH domain interaction can be abolished selectively by point mutations in the putative binding pocket predicted by molecular modelling and NMR spectroscopy. In contrast, the Bruton's tyrosine kinase (Btk)PH domain specifically bound liposomes containing phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3]: an interaction requiring Arg28, a residue found to be mutated in some X-linked agammaglobulinaemia patients. A rational explanation for these different specificities is proposed through modelling of candidate binding pockets and is supported by NMR spectroscopy.
authors.affiliation: Ludwig Institute for Cancer Research, London, UK.
authors.forename: K
authors.initials: K
authors.lastname: Salim
authors.forename: M J
authors.initials: MJ
authors.lastname: Bottomley
authors.forename: E
authors.initials: E
authors.lastname: Querfurth
authors.forename: M J
authors.initials: MJ
authors.lastname: Zvelebil
authors.forename: I
authors.initials: I
authors.lastname: Gout
authors.forename: R
authors.initials: R
authors.lastname: Scaife
authors.forename: R L
authors.initials: RL
authors.lastname: Margolis
authors.forename: R
authors.initials: R
authors.lastname: Gigg
authors.forename: C I
authors.initials: CI
authors.lastname: Smith
authors.forename: P C
authors.initials: PC
authors.lastname: Driscoll
authors.forename: M D
authors.initials: MD
authors.lastname: Waterfield
authors.forename: G
authors.initials: G
authors.lastname: Panayotou
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015374:Biosensing Techniques; D034281:Dynamins; D004789:Enzyme Activation / Q000187:drug effects; D020558:GTP Phosphohydrolases / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D008081:Liposomes; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; C060974:phosphatidylinositol 3,4,5-triphosphate; C037178:phosphatidylinositol 4-phosphate; D011505:Protein-Tyrosine Kinases; D020558:GTP Phosphohydrolases; D034281:Dynamins