title: Distinct specificity in the binding of inositol phosphates by pleckstrin homology domains of pleckstrin, RAC-protein kinase, diacylglycerol kinase and a new 130 kDa protein.
abstract: The pleckstrin homology domains (PH domains) derived from four different proteins, the N-terminal part of pleckstrin, RAC-protein kinase, diacylglycerol kinase and the 130 kDa protein originally cloned as an inositol 1,4,5-trisphosphate binding protein, were analysed for binding of inositol phosphates and derivatives of inositol lipids. The PH domain from pleckstrin bound inositol phosphates according to a number of phosphates on the inositol ring, i.e. more phosphate groups, stronger the binding, but a very limited specificity due to the 2-phosphate was also observed. On the other hand, the PH domains from RAC-protein kinase and diacylglycerol kinase specifically bound inositol 1,3,4,5,6-pentakisphosphate and inositol 1,4,5,6-tetrakisphosphate most strongly. The PH domain from the 130 kDa protein, however, had a preference for inositol 1,4,5-trisphosphate and 1,4,5,6-tetrakisphosphate. Comparison was also made between binding of inositol 1,4,5-trisphosphate, inositol 1,3,4,5-tetrakisphosphate and soluble derivatives of their corresponding phospholipids. The PH domains examined, except that from pleckstrin, showed a 8- to 42-times higher affinity for inositol 1,4,5-trisphosphate than that for corresponding phosphoinositide derivative. However, all PH domains had similar affinity for inositol 1,3,4,5-tetrakisphosphate compared to the corresponding lipid derivative. The present study supports our previous proposal that inositol phosphates and/or inositol lipids could be important ligands for the PH domain, and therefore inositol phosphates/inositol lipids may have the considerable versatility in the control of diverse cellular function. Which of these potential ligands are physiologically relevant would depend on the binding affinities and their cellular abundance.
authors.affiliation: Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Takeuchi
authors.forename: T
authors.initials: T
authors.lastname: Kanematsu
authors.forename: Y
authors.initials: Y
authors.lastname: Misumi
authors.forename: F
authors.initials: F
authors.lastname: Sakane
authors.forename: H
authors.initials: H
authors.lastname: Konishi
authors.forename: U
authors.initials: U
authors.lastname: Kikkawa
authors.forename: Y
authors.initials: Y
authors.lastname: Watanabe
authors.forename: M
authors.initials: M
authors.lastname: Katan
authors.forename: M
authors.initials: M
authors.lastname: Hirata
mesh_terms: D000595:Amino Acid Sequence; D001798:Blood Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D019852:Diacylglycerol Kinase / Q000235:genetics / Q000378:metabolism*; D007295:Inositol Phosphates / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D050356:Lipid Metabolism; D008969:Molecular Sequence Data; D010750:Phosphoproteins; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D011993:Recombinant Fusion Proteins; D017386:Sequence Homology, Amino Acid
chemical_list: D001798:Blood Proteins; D002352:Carrier Proteins; D007295:Inositol Phosphates; D008024:Ligands; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; C039077:platelet protein P47; D019852:Diacylglycerol Kinase; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt