Structural analysis of PLN/SERCA regulatome toward understanding of the mechanism of Ca2+-regulation in the cardiac muscle cells

• Project/Area Number: 25291012
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: The University of Tokyo
• Principal Investigator: Ogawa Haruo 東京大学, 分子細胞生物学研究所, 准教授 (40292726)
• Project Period (FY): 2013-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥18,200,000 (Direct Cost: ¥14,000,000、Indirect Cost: ¥4,200,000); Fiscal Year 2015: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2013: ¥9,620,000 (Direct Cost: ¥7,400,000、Indirect Cost: ¥2,220,000)
• Keywords: 膜蛋白質 / イオンポンプ蛋白質 / X線結晶解析 / X線結晶構造解析

Outline of Final Research Achievements

SR calcium uptake is mediated by a SERCA2a, whose activity is reversibly regulated by its direct mediator phospholamban (PLN). PLN is the most important mediator of β adrenergic signal, and it controls SERCA2a depending on its phosphorylation state. Recently many regulators that controls the PLN/SERCA2a complex was discovered. They are thought to form a large-scale complex (PLN/SERCAa regulatome). However, due to lack of efficient expression/purification system for SERCA2/PLN, the details of the mechanisms how they regulate PLN/SERCAa complex is unclear. Thus, we aimed to elucidate the mechanism of regulation of PLN/SERCA2a by regulators with using our own large expression and purification system.

Research Products

• [Journal Article] Genotype-phenotype correlations of malignant hyperthermia and central core disease mutations in the central region of the RYR1 channel. (2016)
  • Author(s): Murayama, T., Kurebayashi, N., Ogawa, H., Yamazawa, T., Oyamada, H., Suzuki, J., Kanemaru, K., Oguchi, K., Iino, M. and Sakurai, T.
  • Journal Title: Hum Mutat. Volume: 37 Issue: 11 Pages: 1231-1241
  • DOI: 10.1002/humu.23072
  • Peer Reviewed / Open Access
• [Journal Article] Sequential substitution of K+ bound to Na+,K+-ATPase visualized by X-ray crystallography (2015)
  • Author(s): Ogawa, H., Cornelius, F., Hirata, A., Toyoshima, C.
  • Journal Title: Nat Commun. Volume: 6 Issue: 1 Pages: 8004-8004
  • DOI: 10.1038/ncomms9004
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional analysis of SERCA1b, a highly expressed SERCA1 variant in myotonic dystrophy type 1 muscle (2015)
  • Author(s): Zhao, Y., Ogawa, H., Yonekura, S., Mitsuhashi, H., Mitsuhashi, S., Nishino, I., Toyoshima, C., Ishiura, S.
  • Journal Title: Biochim. Biophys. Acta. Volume: 1852 Issue: 10 Pages: 2042-2047
  • DOI: 10.1016/j.bbadis.2015.07.006
  • Peer Reviewed
• [Journal Article] Biselyngbyasides, Cytotoxic Marine Macrolides, are Novel and Potent Inhibitors of the Ca2+ Pumps with a Unique Mode of Binding (2015)
  • Author(s): Morita, M., Ogawa, H., Ohno, O., Yamori, T., Suenaga, K., Toyoshima, C.
  • Journal Title: FEBS Lett. Volume: 589 Issue: 13 Pages: 1406-1411
  • DOI: 10.1016/j.febslet.2015.04.056
  • Peer Reviewed
• [Journal Article] Stimulation, inhibition or stabilization of Na, K-ATPase caused by specific lipid interactions at distinct sites (2015)
  • Author(s): M. Habeck, H. Haviv, A. Katz, E. Kapri-Pardes, S. Ayciriex, A. Shevchenko, H. Ogawa, C. Toyoshima, and S.J.D. Karlish
  • Journal Title: J. Biol. Chem. Volume: 290 Issue: 8 Pages: 4829-4842
  • DOI: 10.1074/jbc.m114.611384
  • Peer Reviewed
• [Journal Article] ナトリウムポンプ蛋白質がナトリウムを選択的に運搬する機構 - How the Na+-pump recognizes and transports Na+ selectively. (2015)
  • Author(s): 小川治夫, 金井隆太, 豊島近
  • Journal Title: 医学のあゆみ Volume: 254 Pages: 1186-1187
• [Journal Article] Crystal structure of a Na^+-bound Na^+, K^+-ATPase preceding the E1P state. (2013)
  • Author(s): R. Kanai, H. Ogawa, B. Vilsen, F. Cornelius and C. Toyoshima
  • Journal Title: Nature Volume: 502 Issue: 7470 Pages: 201-206
  • DOI: 10.1038/nature12578
  • Peer Reviewed
• [Journal Article] Crystal structures of the calcium pump and sarcolipin in the E1·Mg2+ state. (2013)
  • Author(s): C. Toyoshima
  • Journal Title: Nature Volume: 495 Issue: 7440 Pages: 260-264
  • DOI: 10.1038/nature11899
  • Peer Reviewed
• [Presentation] Recent advances in structural studies of SR Ca2+-ATPase (2017)
  • Author(s): 小川治夫
  • Organizer: 第94回日本生理学会
  • Place of Presentation: 浜松アクトシティコングレスセンター（静岡県浜松市）
  • Year and Date: 2017-03-28
  • Invited
• [Presentation] Large-scale production of mammalian membrane proteins toward determination of high resolution structures (2017)
  • Author(s): Haruo Ogawa, Chikashi Toyoshima
  • Organizer: Biophysical Society 61st Annual Meeting
  • Place of Presentation: New Orleans, Louisiana, USA
  • Year and Date: 2017-02-11
  • Int'l Joint Research
• [Presentation] Ca2+-ポンプ構造研究の最前線 (2016)
  • Author(s): 小川治夫、樺島佳樹、豊島近
  • Organizer: 日本生体エネルギー研究会 第42回討論会
  • Place of Presentation: 名古屋工業大学4号館ホール（愛知県名古屋市）
  • Year and Date: 2016-12-19
• [Presentation] 高分解能構造解析へ向けた哺乳類由来膜蛋白質の大量生産 (2016)
  • Author(s): 小川治夫、村山尚、呉林なごみ、豊島近
  • Organizer: 第89回 日本生化学会大会シンポジウム
  • Place of Presentation: 仙台国際センター/東北大学川内北キャンパス（宮城県仙台市）
  • Year and Date: 2016-09-25
  • Invited
• [Presentation] Kinetics by X-ray crystallography:sequential substitution of K+ bound to Na+,K+-ATPase (2016)
  • Author(s): Haruo Ogawa, Flemming Cornelius,Ayami Hirata, Chikashi Toyoshima
  • Organizer: Biophysical Society 60th annual meeting
  • Place of Presentation: Los Angeles Convention center, Los Angeles, California (USA)
  • Year and Date: 2016-02-27
  • Int'l Joint Research
• [Presentation] X線結晶構造解析によるキネティックス測定：Na+,K+-ATPaseに結合したK+は段階的に置換される (2015)
  • Author(s): 小川治夫, 平田絢美, Flemming Cornelius, 豊島近
  • Organizer: 日本生体エネルギー研究会 第41回討論会
  • Place of Presentation: 東京大学医学部１号館講堂（東京都・文京区）
  • Year and Date: 2015-12-21
• [Presentation] Kinetics by X-ray crystallography: sequential substitution of K+ bound to Na+,K+-ATPase. (2015)
  • Author(s): Haruo Ogawa, Flemming Cornelius,Ayami Hirata, Chikashi Toyoshima
  • Organizer: BMB2015（第38回日本分子生物学会 第88回日本生化学会合同大会）ワークショップ
  • Place of Presentation: 神戸ポートアイランド（兵庫県・神戸市）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] Sequential substitution of bound K+ in the transmembrane binding sites of Na+,K+-ATPase, kinetics by X-ray crystallography. (2015)
  • Author(s): Haruo Ogawa, Flemming Cornelius,Ayami Hirata, Chikashi Toyoshima
  • Organizer: 日本生物物理学会第53回年会シンポジウム
  • Place of Presentation: 金沢大学 角間キャンパス自然科学本館（石川県・金沢市）
  • Year and Date: 2015-09-13
  • Invited
• [Presentation] X-線結晶構造解析が明かすビセリングビアサイド類のCa2+ポンプ阻害機構 (2015)
  • Author(s): 森田 真布, 小川 治夫, 杖田 淳子, 大野 修, 矢守 隆夫, 豊島 近, 末永 聖武
  • Organizer: 日本化学会第95春季年会2015
  • Place of Presentation: 日本大学理工学部船橋キャンパス/薬学部（千葉県船橋市）
  • Year and Date: 2015-03-26 – 2015-03-29
• [Presentation] X-ray crystallographic study of Na,K-ATPase in complex with cardiotonic steroids (2015)
  • Author(s): Haruo Ogawa, Kanna Motoyama, Flemming Cornelius, Bente Vilsen, Chikashi Toyoshima
  • Organizer: Biophysical Society 59th Annual meeting
  • Place of Presentation: Baltimore Convention Center, Baltimore, MD, USA
  • Year and Date: 2015-02-07 – 2015-02-11
• [Presentation] Na+,K+-ATPaseと強心配糖体複合体のX線結晶構造解析 (2014)
  • Author(s): 小川 治夫, 元山 かん奈, Cornelius Flemming, Vilsen Bente, 豊島 近
  • Organizer: 第87回日本生化学会
  • Place of Presentation: 京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] 筋強直性ジストロフィーの病態に関わる筋小胞体Ca2+-ATPase 1スプライスバリアントの機能的差異 (2014)
  • Author(s): 趙 一夢, 小川 治夫, 米倉 慎一郎, 三橋 弘明, 豊島 近, 石浦 章一
  • Organizer: 第87回日本生化学会
  • Place of Presentation: 京都国際会館（京都府京都市）
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] Large Production of SERCA toward determination of three-dimensional structures (2014)
  • Author(s): Haruo Ogawa, Yimeng Zhao, Ayami Hirata, Junko Tsueda, Shoichi Ishiura, Giuseppe Inesi, Chikashi Toyoshima
  • Organizer: ASMB special symposia series, Na,K-ATPase and related transport ATPases: Structure, Mechanism, Cell Biology, Health and Disease
  • Place of Presentation: De Werelt Conference Centre, Lunteren, Netherlands
  • Year and Date: 2014-08-30 – 2014-09-05
• [Presentation] Extensive analysis on the interaction between sarcolipin and Ca2+-ATPase (2013)
  • Author(s): Ogawa, H., Iwasawa, S., Hirata, A., Tsueda, J., Toyoshima, C.
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: パシフィコ横浜
• [Presentation] Large production of mammalian membrane proteins toward determination of three-dimensional structures (2013)
  • Author(s): Ogawa, H., Hirata, A., Tsueda, J., Inesi, G., Toyoshima, C.
  • Organizer: The 2nd HD Physiology International Symposium
  • Place of Presentation: 丸の内 My Plaza Hall