title: Design and synthesis of biotinylated inositol 1,3,4,5-tetrakisphosphate targeting Grp1 pleckstrin homology domain.
abstract: A bifunctional molecule containing biotin and d-myo-inositol 1,3,4,5-tetrakisphosphate was synthesized. This molecule was designed on the basis of X-ray structure of the complex of d-myo-inositol 1,3,4,5-tetrakisphosphates, Ins(1,3,4,5)P(4), and Grp1 PH (general receptor of phosphoinositides pleckstrin homology) domain for the application to the widely employed biotin-avidin techniques. The building block of inositol moiety was synthesized starting with myo-inositol and assembled with the biotin-linker moiety through a phosphate linkage. The equilibrium dissociation constant K(D) of biotinylated Ins(1,3,4,5)P(4) binding of original Grp1 PH domain was 0.14 μM in pull-down analysis, which was comparable to that of unmodified Ins(1,3,4,5)P(4). Furthermore, biotinylated Ins(1,3,4,5)P(4) had an ability to distinguish Grp1 PH domain from PLCδ(1) PH domain. Thus, biotinylated Ins(1,3,4,5)P(4) retained the binding affinity and selectivity of original Grp1 PH domain, and realized the intracellular Ins(1,3,4,5)P(4) despite a tethering at the 1-phosphate group of inositol.
authors: Institute of Health Sciences, Kumamoto Health Science University, Kumamoto, Japan.
authors: Kensaku
authors: K
authors: Anraku
authors: Teruhiko
authors: T
authors: Inoue
authors: Kenji
authors: K
authors: Sugimoto
authors: Kota
authors: K
authors: Kudo
authors: Yoshinari
authors: Y
authors: Okamoto
authors: Takashi
authors: T
authors: Morii
authors: Yasuo
authors: Y
authors: Mori
authors: Masami
authors: M
authors: Otsuka
mesh_terms: D000818:Animals; D001710:Biotin / Q000737:chemistry*; D015195:Drug Design; D006801:Humans; D007295:Inositol Phosphates / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
chemical_list: D007295:Inositol Phosphates; D018160:Receptors, Cytoplasmic and Nuclear; C040985:phosphatidylinositol receptors; C081655:inositol-1,3,4,5-tetrakisphosphate; D001710:Biotin