80K-H interacts with inositol 1,4,5-trisphosphate (IP3) receptors and regulates IP3-induced calcium release activity

Katsuhiro Kawaai; Chihiro Hisatsune; Yukiko Kuroda; Akihiro Mizutani; Tomoko Tashiro; Katsuhiko Mikoshiba

doi:10.1074/jbc.M805828200

80K-H interacts with inositol 1,4,5-trisphosphate (IP3) receptors and regulates IP3-induced calcium release activity

J Biol Chem. 2009 Jan 2;284(1):372-380. doi: 10.1074/jbc.M805828200. Epub 2008 Nov 6.

Authors

Katsuhiro Kawaai¹, Chihiro Hisatsune¹, Yukiko Kuroda², Akihiro Mizutani², Tomoko Tashiro², Katsuhiko Mikoshiba³

Affiliations

¹ 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.
² 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.
³ 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.

PMID: 18990696
DOI: 10.1074/jbc.M805828200

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Triphosphate / metabolism
Adenosylhomocysteinase / genetics
Adenosylhomocysteinase / metabolism
Animals
COS Cells
Calcium / metabolism*
Calcium Channels / genetics
Calcium Channels / metabolism*
Calcium Signaling / physiology*
Calcium-Binding Proteins
Cerebellum / cytology
Cerebellum / metabolism
Chlorocebus aethiops
Cytochromes c / genetics
Cytochromes c / metabolism
Endoplasmic Reticulum / genetics
Endoplasmic Reticulum / metabolism
Glucosidases / genetics
Glucosidases / metabolism*
Hippocampus / cytology
Hippocampus / metabolism
Humans
Huntingtin Protein
Inositol 1,4,5-Trisphosphate Receptors / genetics
Inositol 1,4,5-Trisphosphate Receptors / metabolism*
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism*
Lectins, C-Type / genetics
Lectins, C-Type / metabolism
Membrane Proteins / genetics
Membrane Proteins / metabolism
Mice
Mice, Inbred ICR
Microsomes / metabolism
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism
Neurons / cytology
Neurons / metabolism
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Receptors, Cytoplasmic and Nuclear / genetics
Receptors, Cytoplasmic and Nuclear / metabolism*

Substances

CLECL1 protein, human
Calcium Channels
Calcium-Binding Proteins
HTT protein, human
Htt protein, mouse
Huntingtin Protein
ITPR1 protein, human
ITPRIP protein, human
Inositol 1,4,5-Trisphosphate Receptors
Intracellular Signaling Peptides and Proteins
Itpr1 protein, mouse
Lectins, C-Type
Membrane Proteins
Nerve Tissue Proteins
Nuclear Proteins
Prkcsh protein, mouse
Receptors, Cytoplasmic and Nuclear
Adenosine Triphosphate
Cytochromes c
Glucosidases
PRKCSH protein, human
Adenosylhomocysteinase
IRBIT protein, mouse
Calcium