title: 80K-H interacts with inositol 1,4,5-trisphosphate (IP3) receptors and regulates IP3-induced calcium release activity.
abstract: Inositol 1,4,5-trisphosphate receptors (IP3Rs) are intracellular channel proteins that mediate calcium (Ca2+) release from the endoplasmic reticulum, and they are involved in many biological processes (e.g. fertilization, secretion, and synaptic plasticity). Recent reports show that IP3R activity is strictly regulated by several interacting molecules (e.g. IP3R binding protein released with inositol 1,4,5-trisphosphate, huntingtin, presenilin, DANGER, and cytochrome c), and perturbation of this regulation causes intracellular Ca2+ elevation leading to several diseases (e.g. Huntington disease and Alzheimer disease). In this study, we identified protein kinase C substrate 80K-H (80K-H) to be a novel molecule interacting with the COOH-terminal tail of IP3Rs by yeast two-hybrid screening. 80K-H directly interacted with IP3R type 1 (IP3R1) in vitro and co-immunoprecipitated with IP3R1 in cell lysates. Immunocytochemical and immunohistochemical staining revealed that 80K-H colocalized with IP3R1 in COS-7 cells and in hippocampal neurons. We also showed that the purified recombinant 80K-H protein directly enhanced IP3-induced Ca2+ release activity by a Ca2+ release assay using mouse cerebellar microsomes. Furthermore 80K-H was found to regulate ATP-induced Ca2+ release in living cells. Thus, our findings suggest that 80K-H is a novel regulator of IP3R activity, and it may contribute to neuronal functions.
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Kawaai
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Hisatsune
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kuroda
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Mizutani
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Tashiro
authors.affiliation: Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; Department of Chemistry and Biological Science, Aoyama-Gakuin University, 5-10-1 Fuchinobe, Sagamihara, Kanagawa 229-8558, Japan, the Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan, and the Calcium Oscillation Project, International Cooperative Research Project-Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan. Electronic address: mikosiba@brain.riken.jp.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Mikoshiba
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D043383:Adenosylhomocysteinase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002135:Calcium-Binding Proteins; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D002522:Chlorocebus aethiops; D045304:Cytochromes c / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism; D005959:Glucosidases / Q000235:genetics / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D006801:Humans; D000071058:Huntingtin Protein; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008861:Microsomes / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism*
chemical_list: C470184:CLECL1 protein, human; D015220:Calcium Channels; D002135:Calcium-Binding Proteins; C086055:HTT protein, human; C506812:Htt protein, mouse; D000071058:Huntingtin Protein; C506926:ITPR1 protein, human; C516570:ITPRIP protein, human; D053496:Inositol 1,4,5-Trisphosphate Receptors; D047908:Intracellular Signaling Peptides and Proteins; C025389:Itpr1 protein, mouse; D037181:Lectins, C-Type; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; C512949:Prkcsh protein, mouse; D018160:Receptors, Cytoplasmic and Nuclear; D000255:Adenosine Triphosphate; D045304:Cytochromes c; D005959:Glucosidases; C070368:PRKCSH protein, human; D043383:Adenosylhomocysteinase; C510392:IRBIT protein, mouse; D002118:Calcium