Ca2+ and calmodulin-dependent inactivation of the L-type Ca2+ channel and its mutants

• Project/Area Number: 25460294
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General physiology
• Research Institution: Kagoshima University
• Principal Investigator: Minobe Etsuko 鹿児島大学, 医歯学域医学系, 講師 (00448581)
• Research Collaborator: MORI Masayuki 京都大学, 大学院工学研究科, 准教授 (80342640)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: カルシウムチャネル / カルモジュリン / パッチクランプ法 / inside-out / カルシウムイオン / ATP / run-down / リン酸化 / パッチクランプ / カルシウム / Cav1.2

Outline of Final Research Achievements

Calmodulin (CaM) binds to the channel directly and change channel activity in a Ca2+-dependent manner. We reported that the channel activity increases and decreases in a CaM-concentration-dependent manner. Based on our experiments, we propose a model for the regulation of Cav1.2 channel by two CaM binding sites, in which CaM binds to an activation site and then another CaM binds to an inactivation site. To test this hypothesis, we measured the channel activity of C-terminal deletion channel linked with CaM (CaM-linked-channel) and linked with Ca2+-insensitive CaM mutant (CaMmut-linked-channel), using the patch-clamp technique. In the whole-cell recording, the inactivation was diminished in CaMmut-linked-channel. In the inside-out recording, Ca2+-dependent inactivation was observed in the CaM-linked-channel, but not in the CaMmut-linked-channel. The CaM concentration-dependent inactivation was observed in both channels. These results support two-CaM binding site model.

Research Products

• [Journal Article] PKA and phosphatases attached to the Cav1.2 channel regulate channel activity in cell-free patches. (2016)
  • Author(s): Xu JJ, Yu LF, Minobe E, Lu LT, Lei M, Kameyama M
  • Journal Title: Am J Physiol Volume: 310 Issue: 2 Pages: C136-C141
  • DOI: 10.1152/ajpcell.00157.2015
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Regulation of the Cav1.2 cardiac channel by redox via modulation of CaM interaction with the channel. (2015)
  • Author(s): Sun Y, Xu JJ, Minobe E, Shimoara S, Hao LY, Kameyama M
  • Journal Title: J Pharmacol Sci Volume: 128 Issue: 3 Pages: 137-143
  • DOI: 10.1016/j.jphs.2015.06.003
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Nucleotides maintain the activity of Cav1.2 channels in guinea-pig ventricular myocytes. (2015)
  • Author(s): Liu SY, Xu JJ, Minobe E, Gao QH, Feng R, Zhao MM, Guo F, Yang L, Kameyama M, Hao LY
  • Journal Title: Biophys Biochem Res Comm Volume: 460 Issue: 3 Pages: 813-818
  • DOI: 10.1016/j.bbrc.2015.03.111
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Regulation of L-type (CaV1.2) Ca(2+) channels by calmodulin and ATP. (2014)
  • Author(s): Masaki Kameyama, Etsuko Minobe, Jian-Jun Xu, Dong-Yun Han, Hadhimulya Asmara, Asako Kameyama, Rui Feng, Shuyuan Liu, Li-Ying Hao
  • Journal Title: Nihon Yakurigaku Zasshi Volume: 144 Pages: 222-226
  • NAID: 130004704236
• [Journal Article] The individual N- and C-lobes of calmodulin tether to the Cav1.2 channel and rescue the channel activity from run-down in ventricular myocytes of guinea-pig heart. (2014)
  • Author(s): Dongxue Shao, Meimi Zhao, Jianjun Xu, Rui Feng, Feng Guo, Huiyuan Hu, Xuefei Sun, Qingha Gao, Guilin He, Wei Sun, Hongmei Wang, Lifeng Yu, Suyuan Liu, Yaonan Zhu, Etsuko Minobe, Tong Zhu, Masaki Kameyama, Liying Hao
  • Journal Title: FEBS Letters Volume: 588 Issue: 21 Pages: 3855-3861
  • DOI: 10.1016/j.febslet.2014.09.029
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A new phosphorylation site in cardiac L-type Ca2+ channels (Cav1.2) responsible for its cAMP-mediated modulation. (2014)
  • Author(s): Etsuko Minobe, Sachiko Maeda, Jianjun Xu, Liying Hao, Asako Kameyama, Masaki Kameyama
  • Journal Title: American Journal of Physiology Cell Physiology Volume: 307 Issue: 11 Pages: C999-C1009
  • DOI: 10.1152/ajpcell.00267.2014
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Adenosine triphosphate regulates the activity of guinea pig Cav1.2 channel by direct binding to the channel in a dose-dependent manner. (2014)
  • Author(s): Rui Feng, Jianjun Xu, Etsuko Minobe, Asako Kameyama, Lei Yang, Lifeng Yu, Liying Hao, Masaki Kameyama
  • Journal Title: American Journal of Physiology Cell Physiology Volume: 306 Issue: 9 Pages: C856-C863
  • DOI: 10.1152/ajpcell.00368.2013
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The Ca(2+)-dependent interaction of calpastatin domain L with the C-terminal tail of the Cav1.2 channel. (2014)
  • Author(s): Wei Sun, Rui Feng, Hujyuan Hu, Feng Guo, Qinghua Gao, Dongxue Shao, Dandan Yin, Hongmei Wang, Xuefei Sun, Meimi Zhao, Etsuko Minobe, Yu Sun, Guangyu Jiao, Masaki Kameyama, Liying Hao.
  • Journal Title: FEBS Letters Volume: 588 Issue: 5 Pages: 655-671
  • DOI: 10.1016/j.febslet.2014.01.019
  • Peer Reviewed
• [Journal Article] Mechanisms underlying the modulation of L-type Ca2+ channel by hydrogen peroxide in guinea pig ventricular myocytes. (2013)
  • Author(s): Lei Yang, Jianjun Xu, Etsuko Minobe, Li-Feng Yu, Rui Feng, Asako Kameyama, Kazuto Yazawa, Masaki Kameyama.
  • Journal Title: Journal of Physiological Sciences Volume: 3 Issue: 6 Pages: 419-426
  • DOI: 10.1007/s12576-013-0279-2
  • Peer Reviewed
• [Journal Article] Lobe-related concentration- and Ca2+-dependent interactions of calmodulin with C- and N-terminal tails of the CaV1.2 channel. (2013)
  • Author(s): Guilin He, Feng Guo, Zhu T, Dong-Xue Shao, Rui Feng, Yin D, Xuefer Sun, Huiyuan Hu, Hwang A, Etsuko Minobe, Masaki Kameyama, Liying Hao.
  • Journal Title: Journal of Physiological Sciences Volume: 63 Issue: 5 Pages: 345-353
  • DOI: 10.1007/s12576-013-0270-y
  • Peer Reviewed
• [Presentation] Two calmodulin binding site model for the regulation of Cav1.2 channel (2016)
  • Author(s): Kodai Kawaji, Etsuko Minobe, Masayuki X Mori, Masaki Kameyama
  • Organizer: 日本生理学会大会
  • Place of Presentation: 札幌コンベンションセンター（北海道 札幌市）
  • Year and Date: 2016-03-22
• [Presentation] Mechanisms underlying PKA-mediated facilitation of cardiac Cav1.2 channels (2015)
  • Author(s): Liting Lu, Ming Mei, Jianjun Xu, Etsuko Minobe, Kazuto Yazawa, Masaki Kameyama
  • Organizer: 西日本生理学会
  • Place of Presentation: 久留米大学筑水会館（福岡県 久留米市）
  • Year and Date: 2015-10-09
• [Presentation] Cav1.2型チャネルC末端遠位部切断変異体に対するカルモジュリンとATPの作用 (2015)
  • Author(s): 蓑部悦子、森誠之、亀山正樹
  • Organizer: 西日本生理学会
  • Place of Presentation: 久留米大学筑水会館（福岡県 久留米市）
  • Year and Date: 2015-10-09
• [Presentation] PKA-mediated facilitation of cardiac Cav1.2 channel through uncovering calmodulin binding sites by distal C-terminus of α1c subunit. (2015)
  • Author(s): Jianjun Xu, Etsuko Minobe, Yu Sun, Liting Lu, Ming Lei, Kazuto Yazawa, Masaki Kameyama
  • Organizer: 日本生理学会大会
  • Place of Presentation: 兵庫 (神戸国際会議場・展示場）
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] Regulation of cardiac Cav1.2 channel by redox via modulation of CaM interaction with channel (2015)
  • Author(s): Yu Sun, Jianjun Xu, Etsuko Minobe, Shoken, Shimoara, Liying Hao, Masaki Kameyama
  • Organizer: 日本生理学会大会
  • Place of Presentation: 兵庫 (神戸国際会議場・展示場）
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] Lobe-specific interactions of calmodulin with C-terminal tails of cardiac CaV1.2 channel. (2014)
  • Author(s): Dongxue Shao, Jianjun Xu, Meimi Zhao, Etsuko Minobe,Liying Hao, Masaki Kameyamao
  • Organizer: 西日本生理学会
  • Place of Presentation: 沖縄 (琉球大学研究者交流施設50周年記念館）
  • Year and Date: 2014-10-23 – 2014-10-24
• [Presentation] Cav1.2Ca2+チャネルとCaMの相互作用の分子機構 (2014)
  • Author(s): Dongxue Shao, Meimi Zhao, 何桂林、郭鳳、呂力婷、蓑部悦子、徐建軍、亀山亜砂子、Tong Zhu、Liying Hao、亀山正樹
  • Organizer: 生理学研究所研究会
  • Place of Presentation: 愛知 (自然科学研究機構生理学研究所）
  • Year and Date: 2014-09-04 – 2014-09-05
• [Presentation] Regulation and molecular mechanisms of Cav1.2 channel by calmodulin and ATP (2014)
  • Author(s): Estuko Minobe, Dong-Yun Han, Hadhimulya Asmara, Rui Feng, Jianjun Xu, Masaki Kameyama
  • Organizer: 日本生理学会大会
  • Place of Presentation: 鹿児島
• [Presentation] The role of nucleotides in regulation of cardiac Cav1.2 channel (2014)
  • Author(s): Shuyuan Liu, Jianjyn Xu, Etsuko Minobe, Asako Kameyama, Liying Hao, Masaki Kameyama
  • Organizer: 日本生理学会大会
  • Place of Presentation: 鹿児島
• [Presentation] Regulation of L-type Cav1.2 Ca2+ chanels by ATP (2014)
  • Author(s): Masaki Kameyama, Rui Feng, Shunyuan Liu, Etsuko Minobe, Jianjun Xu, Asako Kameyama, Hao Liying
  • Organizer: 日本生理学会大会
  • Place of Presentation: 鹿児島
• [Presentation] The lobe-specific interaction of calmodulin with the cardiac CaV1.2 channel (2014)
  • Author(s): Dong-Xue Shao, Jianjun Xu, Meimi Ahao, Yu Sun, Etsuko Minobe, Guilin, Rui Feng, Sun Xuefer, Feng Guo、Masaki Kameyama, Liying Hao
  • Organizer: 日本生理学会大会
  • Place of Presentation: 鹿児島
• [Presentation] Regulation of L-type (Cav1.2) Ca2+ channes by calmodulin and ATP. (2014)
  • Author(s): Masaki Kameyama, Etsuko Minobe, Jianjun Xu, Dongyun Han, Hadmulya Asmara, Asako Kameyama, Rui Feng, Shuyan Liu, Liying Hao
  • Organizer: 日本薬理学会年会
  • Place of Presentation: 宮城
• [Presentation] Properties of the ATP-binding site responsible for regulation of cardiac Ca2+ channels. (2013)
  • Author(s): Rui Feng, Shuyuan Liu, Jian-Jun Xu, Etsuko Minobe, Asako Kameyama, Li-Ying Hao, Masaki Kameyama
  • Organizer: 西日本生理学会
  • Place of Presentation: 福岡
• [Remarks] 鹿児島大学大学院医歯学総合研究科神経筋生理学講座
  • URL: http://www.kufm.kagoshima-u.ac.jp/^physiol2/
• [Remarks] イオンチャネルに作用する薬物と毒物
  • URL: http://www.rdc.kagoshima-u.ac.jp/rdc/search/upload/kameyama-med.pdf
• [Remarks] 鹿児島大学大学院医歯学総合研究科神経筋生理学講座
  • URL: http://www.kufm.kagoshima-u.ac.jp/~physiol2/
• [Remarks] イオンチャンネルに作用する薬物と毒物
  • URL: http://www.rdc.kagoshima-u.ac.jp/rdc/search/upload/kameyama-med.pdf
• [Remarks] http://www.kufm.kagoshima-u.ac.jp/~physiol2/