title: Structure and phosphatidylinositol-(3,4)-bisphosphate binding of the C-terminal PH domain of human pleckstrin.
abstract: Pleckstrin is the major target of protein kinase C (PKC) in blood platelets. Its phosphorylation triggers responses that ultimately lead to platelet activation and blood clot formation. Pleckstrin consists of three domains: a pleckstrin homology (PH) domain at both termini and a central DEP (Dishevelled, Egl-1, Pleckstrin) domain. Here, we report the solution nuclear magnetic resonance (NMR) structure of the C-terminal PH domain (C-PH) of human pleckstrin-1. We show that this PH domain binds phosphatidylinositol-3,4-bisphosphate (PtdIns(3,4)P2) with high specificity in protein lipid overlay assays. Using NMR titration experiments and mutational analysis, residues involved in binding to PtdIns(3,4)P2 are identified. The binding site is formed by a patch of basic residues from the beta1 and beta2 strands and the beta1-beta2 loop. Since PtdIns(3,4)P2 is an important signaling molecule in platelets, our data suggest a C-PH dependent regulation of pleckstrin function in response to PtdIns(3,4)P2.
authors.affiliation: EMBL Heidelberg, Meyerhofstr. 1, 69117 Heidelberg, Germany. edlich@embl.de
authors.forename: Christian
authors.initials: C
authors.lastname: Edlich
authors.forename: Gunter
authors.initials: G
authors.lastname: Stier
authors.forename: Bernd
authors.initials: B
authors.lastname: Simon
authors.forename: Michael
authors.initials: M
authors.lastname: Sattler
authors.forename: Claudia
authors.initials: C
authors.lastname: Muhle-Goll
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D001798:Blood Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary
chemical_list: D001798:Blood Proteins; D008024:Ligands; D018129:Phosphatidylinositol Phosphates; D010750:Phosphoproteins; C060980:phosphatidylinositol 3,4-diphosphate; C039077:platelet protein P47