ID | 57503 |
フルテキストURL | |
著者 |
Takabatake, Shota
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Ohtsuka, Satomi
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Sugawara, Takeyuki
Department of Anatomy, Kitasato University School of Medicine
Hatano, Naoya
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Kanayama, Naoki
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
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Magari, Masaki
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Sakagami, Hiroyuki
Department of Anatomy, Kitasato University School of Medicine
Tokumitsu, Hiroshi
Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
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抄録 | BACKGROUND:
Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) is a pivotal activator of CaMKI, CaMKIV and 5'-AMP-activated protein kinase (AMPK), controlling Ca2+-dependent intracellular signaling including various neuronal, metabolic and pathophysiological responses. Recently, we demonstrated that CaMKKβ is feedback phosphorylated at Thr144 by the downstream AMPK, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme. However, the regulatory phosphorylation of CaMKKβ at Thr144 in intact cells and in vivo remains unclear. METHODS: Anti-phosphoThr144 antibody was used to characterize the site-specific phosphorylation of CaMKKβ in immunoprecipitated samples from mouse cerebellum and in transfected mammalian cells that were treated with various agonists and protein kinase inhibitors. CaMKK activity assay and LC-MS/MS analysis were used for biochemical characterization of phosphorylated CaMKKβ. RESULTS: Our data suggest that the phosphorylation of Thr144 in CaMKKβ is rapidly induced by cAMP/cAMP-dependent protein kinase (PKA) signaling in CaMKKβ-transfected HeLa cells, that is physiologically relevant in mouse cerebellum. We confirmed that the catalytic subunit of PKA was capable of directly phosphorylating CaMKKβ at Thr144 in vitro and in transfected cells. In addition, the basal phosphorylation of CaMKKβ at Thr144 in transfected HeLa cells was suppressed by AMPK inhibitor (compound C). PKA-catalyzed phosphorylation reduced the autonomous activity of CaMKKβ in vitro without significant effect on the Ca2+/CaM-dependent activity, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme. CONCLUSION: cAMP/PKA signaling may confer Ca2+-dependency to the CaMKKβ-mediated signaling pathway through direct phosphorylation of Thr144 in intact cells. GENERAL SIGNIFICANCE: Our results suggest a novel cross-talk between cAMP/PKA and Ca2+/CaM/CaMKKβ signaling through regulatory phosphorylation. |
キーワード | CaMKK
Calmodulin
Intracellular Ca(2+)
PKA
Phosphorylation
Signal transduction
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発行日 | 2019-01-17
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出版物タイトル |
Biochimica et Biophysica Acta (BBA) - General Subjects
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巻 | 1863巻
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号 | 4号
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出版者 | Elsevier Science
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開始ページ | 672
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終了ページ | 680
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ISSN | 0304-4165
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NCID | AA00564679
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資料タイプ |
学術雑誌論文
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言語 |
英語
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OAI-PMH Set |
岡山大学
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論文のバージョン | author
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PubMed ID | |
DOI | |
Web of Science KeyUT | |
関連URL | isVersionOf https://doi.org/10.1016/j.bbagen.2018.12.012
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Citation | Shota Takabatake, Satomi Ohtsuka, Takeyuki Sugawara, Naoya Hatano, Naoki Kanayama, Masaki Magari, Hiroyuki Sakagami, Hiroshi Tokumitsu, Regulation of Ca2+/calmodulin-dependent protein kinase kinase β by cAMP signaling, Biochimica et Biophysica Acta (BBA) - General Subjects, Volume 1863, Issue 4, 2019, Pages 672-680, ISSN 0304-4165, https://doi.org/10.1016/j.bbagen.2018.12.012.
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助成機関名 |
日本学術振興会
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助成番号 | 18K06113
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