title: A comparative analysis of the phosphoinositide binding specificity of pleckstrin homology domains.
abstract: Pleckstrin homology (PH) and phosphotyrosine binding (PTB) domains are structurally related regulatory modules that are present in a variety of proteins involved in signal transduction, such as kinases, phospholipases, GTP exchange proteins, and adapter proteins. Initially these domains were shown to mediate protein-protein interactions, but more recently they were also found to bind phosphoinositides. Most studies to date have focused on binding of PH domains to phosphatidylinositol (PtdIns)-4-P and PtdIns-4,5-P2 and have not considered the lipid products of phosphoinositide 3-kinase: PtdIns-3-P, PtdIns-3,4-P2, and PtdIns-3,4,5-P3. Here we have compared the phosphoinositide specificity of six different PH domains and the Shc PTB domain using all five phosphoinositides. We show that the Bruton's tyrosine kinase PH domain binds to PtdIns-3,4, 5-P3 with higher affinity than to PtdIns-4,5-P2, PtdIns-3,4-P2 or inositol 1,3,4,5-tetrakisphosphate (Ins-1,3,4,5-P4). This selectivity is decreased by the xid mutation (R28C). Selective binding of PtdIns-3,4,5-P3 over PtdIns-4,5-P2 or PtdIns-3,4-P2 was also observed for the amino-terminal PH domain of T lymphoma invasion and metastasis protein (Tiam-1), the PH domains of Son-of-sevenless (Sos) and, to a lesser extent, the PH domain of the beta-adrenergic receptor kinase. The oxysterol binding protein and beta-spectrin PH domains bound PtdIns-3,4,5-P3 and PtdIns-4,5-P2 with similar affinities. PtdIns-3,4,5-P3 and PtdIns-4,5-P2 also bound to the PTB domain of Shc with similar affinities and lipid binding was competed with phosphotyrosine (Tyr(P)-containing peptides. These results indicate that distinct PH domains select for different phosphoinositides.
authors.affiliation: Department of Cell Biology, Harvard Medical School and Division of Signal Transduction, Beth Israel Hospital, Boston, Massachusetts 02115, USA.
authors.forename: L E
authors.initials: LE
authors.lastname: Rameh
authors.forename: A k
authors.initials: Ak
authors.lastname: Arvidsson
authors.forename: K L
authors.initials: KL
authors.lastname: Carraway
authors.forename: A D
authors.initials: AD
authors.lastname: Couvillon
authors.forename: G
authors.initials: G
authors.lastname: Rathbun
authors.forename: A
authors.initials: A
authors.lastname: Crompton
authors.forename: B
authors.initials: B
authors.lastname: VanRenterghem
authors.forename: M P
authors.initials: MP
authors.lastname: Czech
authors.forename: K S
authors.initials: KS
authors.lastname: Ravichandran
authors.forename: S J
authors.initials: SJ
authors.lastname: Burakoff
authors.forename: D S
authors.initials: DS
authors.lastname: Wang
authors.forename: C S
authors.initials: CS
authors.lastname: Chen
authors.forename: L C
authors.initials: LC
authors.lastname: Cantley
mesh_terms: D000077329:Agammaglobulinaemia Tyrosine Kinase; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001798:Blood Proteins / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010750:Phosphoproteins; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D011987:Receptors, Steroid / Q000378:metabolism; D020664:Son of Sevenless Proteins; D013049:Spectrin / Q000378:metabolism; D000098825:Oxysterol Binding Proteins
chemical_list: D001798:Blood Proteins; D008565:Membrane Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010750:Phosphoproteins; D011506:Proteins; D011987:Receptors, Steroid; D020664:Son of Sevenless Proteins; D000098825:Oxysterol Binding Proteins; C060974:phosphatidylinositol 3,4,5-triphosphate; C060980:phosphatidylinositol 3,4-diphosphate; C039077:platelet protein P47; D013049:Spectrin; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases; D000077329:Agammaglobulinaemia Tyrosine Kinase