Molecular Mechanism of Target Recognition by Calmodulin

钙调蛋白识别靶点的分子机制

• 批准号: 10450358
• 负责人: IZUMI Yoshinobu
• 金额: $ 9.02万
• 依托单位: Yamagata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10450358/
• 关键词: Calmodulin; Myosin light chain kinase; Calcineurin; Calmodulin-dependent protein kinase I, II, IV; CAP-23; NAP-22; W-7; TFP; Recognition Sequence; 標準的標的分子; 非標準的標的分子; カルモデュリン依存性プロテインキナーゼ; アポカルモデュリン; NAPIZZ; 標的ペプチド; 分子認識モチーフ; 放射光小角散乱; 多機能性発

9-1) Small-angle X-ray scattering (SAXS) was used to investigate the conformational changes of calmodulin on binding to a calmodulin-binding sequence in myosin light chain kinase, calcineurin, calmodulin-protein kinase. I, II, IV and these binding motifs were determined on the molecular level.9-2) SAXS was used to investigate the conformational changes of calmodulin on binding to a myristoylated N-terminal nonapeptide from neuron-specific protein CAP-23/NAP-22 and the binding motif was determined on the molecular level.9-3) SAXS was used to investigate the conformational changes of calmodulin on binding to an antagonist like W-7 and TFP and the binding motif was determined on the molecular level.9-4) In the absence of Ca^<2+>, a new binding motif was found in the calmodulin/TFP and calmodulin/calmodulin-dependent protein kinase TV complexes.9-5) SAXS was used to reveal the role of the central helix in calmodulin for its recognition.9-6) SAXS with stopped-flow technique was used to reveal the kinetic conformational change of Ca^<2+>-dependent target recognition by calmodulin.

9-1) 使用小角X射线散射(SAXS)来研究钙调蛋白与肌球蛋白轻链激酶、钙调神经磷酸酶、钙调蛋白蛋白激酶中的钙调蛋白结合序列结合时的构象变化。 I、II、IV 和这些结合基序在分子水平上确定。9-2) 使用 SAXS 研究钙调蛋白与神经元特异性蛋白 CAP-23/NAP- 的肉豆蔻酰化 N 末端九肽结合时的构象变化。 22并在分子水平上确定了结合基序。9-3) 使用SAXS研究了钙调蛋白与W-7和TFP等拮抗剂结合时的构象变化，并确定了结合基序在分子水平上测定。9-4)在Ca ^ 2+ 不存在的情况下，在钙调蛋白/TFP和钙调蛋白/钙调蛋白依赖性蛋白激酶TV复合物中发现了新的结合基序。9-5)使用SAXS揭示了中央螺旋在钙调蛋白识别中的作用。9-6)采用停流技术的SAXS技术揭示了钙调蛋白对Ca^2+依赖性靶标识别的动力学构象变化。

项目成果

Y.Izumi, H.Takezawa, N.Kikuta, S.Uemura, A.Tsutsumi: "Two-Stage Melting in Dilute Gels of poly (g-benzyl L-glutamate)"Macromlecules. 31. 430-435 (1998)

Y.Izumi、H.Takezawa、N.Kikuta、S.Uemura、A.Tsutsumi：“聚（g-苄基L-谷氨酸）稀凝胶中的两阶段熔化”大分子。

M.Osawa, S.Kuwamoto, Y.Izumi 他: "Evidence for calmodulin inter-domain compaction in solusion induced by W-7 binding" FEBs Lett.442. 173-177 (1999)

M.Osawa、S.Kuwamoto、Y.Izumi 等人：“W-7 结合诱导的溶液中钙调蛋白域间压缩的证据”FEBs Lett.442 (1999)。

N.Matsushima,N.hayachi,Y.Jinbo.Y-lzumi: "Ca^<2+>-bound calmodulin forms a compact globular structrue on binding four trifluoperazine molecules in solution"Biochem.J.. 347. 211-215 (2000)

N.Matsushima，N.hayachi，Y.Jinbo.Y-lzumi：“Ca^2-结合的钙调蛋白在溶液中结合四个三氟拉嗪分子时形成致密的球状结构”Biochem.J.. 347. 211-215 (2000

M.Osawa, S.Kuwamoto, Y.Izumi, K.L.Yorp, M.Ikura, N.Matsusima: "Evidenae for calmodulin inter-dowain compaition in solution"FEBS Letters. 442. 173-177 (1999)

M.Osawa、S.Kuwamoto、Y.Izumi、K.L.Yorp、M.Ikura、N.Matsusima：“溶液中钙调蛋白域间竞争的证据”FEBS 快报。

N.Matsushima, N.Hayasi, Y.Jinbo, Y.Izumi.: "Ca^<2+>-bound calmodulin forms a compact globular structure on binding four trifluoperazine molecules in solution"Biochem.J.. 347. 211-215 (2000)

N.Matsushima、N.Hayasi、Y.Jinbo、Y.Izumi.：“Ca^2-结合的钙调蛋白在溶液中结合四个三氟拉嗪分子时形成致密的球状结构”Biochem.J.. 347. 211-215 (