Analysis of a newly cloned stretch-activated ion channel from cardiac cells

分析新克隆的心肌细胞拉伸激活离子通道

• 批准号: 13470009
• 负责人: NARUSE Keiji
• 金额: $ 6.08万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470009/
• 关键词: SA channel; Ca-activated K chanel; SAKCA; structure-function relationship; Stretch-activated channel; 心筋; 伸展刺激; パッチクランプ法

Stretch-sensing mechanisms of stretch-activated (SA) channels have been extensively studied and many molecular models have been presented. Here we cloned a stretch-activated, large-conductance (BK) channel from cultured chick embryonic heart and found out that the channel contains "STREX" sequence, originally reported to be a target sequence of PKA and whose expression is regulated by stress hormones, and that it turned out to contain a mechano-sensive domain of the channel. Deletion of STREX or substitution of Ala^<674> to Thr^<674> diminished stretch-sensitivity of the channel. When compared the chick STREX with mouse, rabbit STREX, which did not have any stretch-sensitivity in CHO cells, it turned out that the sequence ERA is most likely to be important. When chick STREX-EGFP was co-expressed with SAKCA in CHO cells, a strong signal was detected at cell membrane and stretch-sensitivity of the channel was significantly inhibited.

牵张激活（stretch-activated，SA）通道的牵张敏感机制已被广泛研究，并提出了许多分子模型。本研究从鸡胚心脏中克隆了一个牵张激活的大电导（BK）通道，并发现该通道含有“STREX”序列，该序列最初被报道为PKA的靶序列，其表达受应激激素的调节，而且该通道含有一个机械敏感结构域。缺失STREX或将Ala^替换<674>为Thr^会<674>降低通道的拉伸敏感性。当将鸡STREX与在CHO细胞中没有任何拉伸敏感性的小鼠、兔STREX进行比较时，结果表明ERA序列最有可能是重要的。当鸡STREX-EGFP与SAKCA在CHO细胞中共表达时，在细胞膜上检测到强信号，并且通道的拉伸敏感性被显著抑制。

项目成果

Wang N, Naruse K, Stamenovic O, Fredberg JJ, M SM, Tolic-Norrelykke IM, Polte T, Mannix R: "Mechanical behavior in living cells consistent with the tensegrity model"Proc Natl Acad Sci USA. 98. 7765-7770 (2001)

Wang N、Naruse K、Stamenovic O、Fredberg JJ、M SM、Tolic-Norrelykke IM、Polte T、Mannix R：“活细胞中的机械行为与张拉整体模型一致”Proc Natl Acad Sci USA。

Kawal M, Naruse K, Komatsu S, Kobayashi S, Nagino M, Nimura Y, Sokabe M.: "Mechanical stress-dependent secretion of interleukin 6 by endothelial cells after portal vein embolization : clinical and experimental studies"J Hepatol. 37. 240-246 (2002)

Kawal M、Naruse K、Komatsu S、Kobayashi S、Nagino M、Nimura Y、Sokabe M.：“门静脉栓塞后内皮细胞机械应激依赖性白介素 6 分泌：临床和实验研究”J Hepatol。

Wang JG, Miyazu M, Matsushita E, Sokabe M, Naruse K: "Uniaxial cyclic stretch induces focal adhesion kinase (FAK) tyrosine phosphorylation followed by mitogen-activated protein kinase (MAPK) activation"Biochem Biophys Res Commun. 288. 356-361 (2001)

Wang JG、Miyazu M、Matsushita E、Sokabe M、Naruse K：“单轴循环拉伸诱导粘着斑激酶 (FAK) 酪氨酸磷酸化，随后激活丝裂原激活蛋白激酶 (MAPK)”Biochem Biophys Res Commun。

S.Takayama, E.Ostuni, P.LeDuc, K.Naruse, D.Ingber, G.Whitesides: "Selective Chemical Treatment of Cellular Microdomains Using Multiple Laminar Streams"Chemistry and Biology. (印刷中).

S. Takayama、E. Ostuni、P. LeDuc、K. Naruse、D. Ingber、G. Whitesides：“使用多重层流对细胞微域进行选择性化学处理”化学与生物学（正在出版）。

Aikawa K: "SA channel mediates superoxide production in HUVECs"Life Sciences. 69. 1717-1724 (2001)

Aikawa K：“SA 通道介导 HUVEC 中超氧化物的产生”生命科学。