1995 Fiscal Year Annual Research Report

カルシウム/カルモデュリン依存性多機能性タンパク質リン酸化酵素の活性制御機構

Research Project

Project/Area Number	07457578
Research Institution	Asahikawa Medical College
Principal Investigator	藤澤仁旭川医科大学, 医学部, 教授 (10027039)
Co-Investigator(Kenkyū-buntansha)	竹内昌之旭川医科大学, 医学部, 助手 (40226999) 石田敦彦旭川医科大学, 医学部, 助手 (90212886) 亀下勇旭川医科大学, 医学部, 助教授 (60127941)
Keywords	カルシウム / カルモデュリン / 脳 / タンパク質リン酸化酵素 / シグナル伝達 / 酵素活性の調節機構 / タンパク質リン酸化反応 / 阻害剤
Research Abstract	1.カルシウム/カルモデュリン依存性タンパク質リン酸化酵素II(CaMキナーゼII) 1)CaMキナーゼIIの特異的な活性阻害ペプチド(AIP)を開発し、これまで市販され使われてきた阻害剤、KN-93などと比べると桁違いに強力且つ特異的な阻害剤であることを示した。 2)上記AIPを用いて膵臓からのインスリン分泌に関与するサイクリックADPリボースによる小胞体からのカルシウム遊離にCaMキナーゼIIが作用していることを明らかにした。 3)CaMキナーゼIIの自己活性抑制ドメインは酵素を不活性化しているだけでなく酵素の安定化もしていることを示し、その機能について調べた。 4)CaMキナーゼIIのスレオニン^<28.6>の自己リン酸化がCaMキナーゼIIの著明な活性化を引き起こすという我々の考えを証明した。 2.カルシウム/カルモデュリン依存性タンパク質リン酸化酵素IV(CaMキナーゼIV) 1)CaMキナーゼIVの活性発現のために必要なカルシウム/カルモデュリンは酵素の不活性化作用も併せ持っていることを示した。 2)CaMキナーゼIVはCaMキナーゼIVキナーゼによって一個のリン酸化を受けて活性化されるが、その後カルシウムイオン濃度が低下すると、セリン^<332>を自己リン酸化してカルシウム/カルモデュリン依存性活性を失うことを明らかにした。 3)CaMキナーゼIVキナーゼをラットの脳からクローニングし明らかにしたアミノ酸配列から特異抗体を調製した。 4)CaMキナーゼおよびCaMキナーゼIVキナーゼの脳内分布、細胞内分布を免疫組織学的に調べ、両者の分布を比較検討した。

Research Products
(13 results)

All Other

All Publications (13 results)

[Publications] I.Kameshita: "Preparation and Characterization of Calmodulin-dependent Protein Kinase IV (CaM-kinase IV) free of CaM-kinase IV Kinase from Rat Cerebral Cortex" J.Biochem.117. 85-90 (1995)
[Publications] A.Ishida: "Stabilization of Calmodulin-dependent Protein Kinase II through the Autoinhibitory Domain" J.Biol.Chem.270. 2163-2170 (1995)
[Publications] M.Takeuchi: "Contact Transfer Method : DNA transfer from polyacrylamide gel" Anal.Biochem.224. 611-612 (1995)
[Publications] Y.Nakamura: "An Immunohistochemical study of Ca^<2+>/Calmodulin-dependent Protein Kinase IV in the Rat Central Nervous system : Light and Electron Microscopic Observations" Neuroscience. 68. 181-194 (1995)
[Publications] S.Okuno: "Full Activation of Brain Calmodulin-Dependent Protein Kinase IV Requires Phosphorylation of the Amino-Terminal Serine-Rich Region by Calmodulin-Dependent Protein Kinase IV Kinase" J.Biochem.117. 686-690 (1995)
[Publications] T.Kitani: "Inactivation of Ca^<2+>/Calmodulin-Dependent Protein Kinase IV by Ca^<2+>/Calmodulin and Restoration of the Activity by Mg^<2+>/EGTA" J.Biochem.117. 1070-1075 (1995)
[Publications] A.Ishida: "A Novel Highly Specific and Potent Inhibitor of Calmodulin-Dependent Protein Kinase II" Biochem.Biophys.Res.Commun.212. 806-812 (1995)
[Publications] T.Kitani: "Comparison of Ca^<2+>/Calmodulin-Dependent Protein Kinase IV from Rat Brain,Expressed in Insect Cells,and Expressed in Escherichia coli" J.Biochem.118. 364-370 (1995)
[Publications] S.Takasawa: "Requirement of calmodulin-dependent protein kinase II in cyclic ADP-ribose-mediated intracellular Ca^<2+> mobilization" J.Biol.Chem.270. 30257-30259 (1995)
[Publications] K.Yoshida: "Differential Distribution fo CaM Kinases and Induction of c-fos Expression by Flashing and Sustained Light in Rat Retinal Cells" Inv.Ophthalmol.Vis.Sci.(in press).
[Publications] A.Ishida: "Evidence that autophosphorylation at Thr-286/Thr-287 is required for full activation of calmodulin-dependent protein kinase II" Biochim.Biophys.Acta. (in press).
[Publications] Y.Nakamura: "Distribution of Ca^<2+>/calmodulin-dependent protein kinase kinase α in the rat central nervous system : an immunohistochemical study" Neuroscience Letters. (in press).
[Publications] S.Watanabe: "Inactivation of calmodulin-dependent protein kinase IV by autophosphorylation of serine 332 within the putative calmodulin-binding domain" J.Biol.Chem.(in press).

1995 Fiscal Year Annual Research Report

カルシウム/カルモデュリン依存性多機能性タンパク質リン酸化酵素の活性制御機構

Principal Investigator

藤澤 仁 旭川医科大学, 医学部, 教授 (10027039)

Research Products

[Publications] I.Kameshita: "Preparation and Characterization of Calmodulin-dependent Protein Kinase IV (CaM-kinase IV) free of CaM-kinase IV Kinase from Rat Cerebral Cortex" J.Biochem.117. 85-90 (1995)

[Publications] A.Ishida: "Stabilization of Calmodulin-dependent Protein Kinase II through the Autoinhibitory Domain" J.Biol.Chem.270. 2163-2170 (1995)

[Publications] M.Takeuchi: "Contact Transfer Method : DNA transfer from polyacrylamide gel" Anal.Biochem.224. 611-612 (1995)

[Publications] Y.Nakamura: "An Immunohistochemical study of Ca^<2+>/Calmodulin-dependent Protein Kinase IV in the Rat Central Nervous system : Light and Electron Microscopic Observations" Neuroscience. 68. 181-194 (1995)

[Publications] S.Okuno: "Full Activation of Brain Calmodulin-Dependent Protein Kinase IV Requires Phosphorylation of the Amino-Terminal Serine-Rich Region by Calmodulin-Dependent Protein Kinase IV Kinase" J.Biochem.117. 686-690 (1995)

[Publications] T.Kitani: "Inactivation of Ca^<2+>/Calmodulin-Dependent Protein Kinase IV by Ca^<2+>/Calmodulin and Restoration of the Activity by Mg^<2+>/EGTA" J.Biochem.117. 1070-1075 (1995)

[Publications] A.Ishida: "A Novel Highly Specific and Potent Inhibitor of Calmodulin-Dependent Protein Kinase II" Biochem.Biophys.Res.Commun.212. 806-812 (1995)

[Publications] T.Kitani: "Comparison of Ca^<2+>/Calmodulin-Dependent Protein Kinase IV from Rat Brain,Expressed in Insect Cells,and Expressed in Escherichia coli" J.Biochem.118. 364-370 (1995)

[Publications] S.Takasawa: "Requirement of calmodulin-dependent protein kinase II in cyclic ADP-ribose-mediated intracellular Ca^<2+> mobilization" J.Biol.Chem.270. 30257-30259 (1995)

[Publications] K.Yoshida: "Differential Distribution fo CaM Kinases and Induction of c-fos Expression by Flashing and Sustained Light in Rat Retinal Cells" Inv.Ophthalmol.Vis.Sci.(in press).

[Publications] A.Ishida: "Evidence that autophosphorylation at Thr-286/Thr-287 is required for full activation of calmodulin-dependent protein kinase II" Biochim.Biophys.Acta. (in press).

[Publications] Y.Nakamura: "Distribution of Ca^<2+>/calmodulin-dependent protein kinase kinase α in the rat central nervous system : an immunohistochemical study" Neuroscience Letters. (in press).

[Publications] S.Watanabe: "Inactivation of calmodulin-dependent protein kinase IV by autophosphorylation of serine 332 within the putative calmodulin-binding domain" J.Biol.Chem.(in press).

藤澤仁旭川医科大学, 医学部, 教授 (10027039)