SCREENING FOR THE FUNCTIONAL DOMAINS IN THE RYANODINE RECEPTOR

兰尼碱受体功能域的筛选

• 批准号: 15590234
• 负责人: OGUCHI Katsuji
• 金额: $ 2.24万
• 依托单位: SHOWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590234/
• 关键词: calcium ion; calcium release channel; ryanodine receptor; green fluorescent protein(GFP); endoplasmic reticulum; 部分領域; 部分欠損クローン

1.We divided the ryanodine receptor (RyR1) cDNA (about 15000bp) from a rabbit into 11 cassettes by introducing unique restriction endonuclease sites (5'-XbaI, 677SalI, 2349Bsu36I, 3798BstBI, 5355MluI, 6822SpeI, 8476BsiWI, 9804AvrII, 11290NdeI, 12675NheI, 14427ClaI, and edge-HindIII-3'end).2.We expressed the recombinant RyR1 mutants encoded by partly deleted clones (16 clones) or by partial clones (12 clones) reconstructed by the divided cassettes fused with a green fluorescent protein (GFP) in CHO cells.3.The GFP fluorescent signals from the CHO cells transfected with the cassettes 1,5,6,7 or 11 were observed as the dot-like structures in the cytoplasm except for the nucleus and could be remained even after the plasma membranes permeabilized as seen in those cells having the full-length of RyR1 expressed.4.These results reveals that there are several regions (the amino terminal, the central and the carboxyl terminal regions) of the RyR1 molecule which are responsible for the retention / retrieval signals to the endoplasmic reticulum as the intracellular calcium stores of the cells. These regions for the ER retention are corresponded to the genetic mutations of RyR1 clustered in the malignant hyperthermia patients also suggesting that these regions have the important roles for the calcium release channel

1.我们克隆了ryanodine受体（RyR 1）cDNA，通过引入独特的限制性内切酶位点，将来自兔的约15000 bp的片段插入11个盒中（5'-XbaI，677SalI，2349Bsu36I，3798BstBI，5355MluI，6822SpeI，8476BsiWI，9804AvrII，11290NdeI，12675NheI，14427ClaI，2.表达了部分缺失的RyR 1突变体（16个克隆）或部分克隆（12个克隆）。3.来自转染有盒1，5，6，7或11在细胞质中观察到除了细胞核之外的点状结构，并且即使在质膜透化之后也可以保留，如在具有全长RyR 1表达的那些细胞中所观察到的。（氨基末端，RyR 1分子的中心和羧基末端区域），其负责保留/回收信号传递到内质网作为细胞的细胞内钙储存。这些ER滞留区域与恶性高热患者聚集的RyR 1基因突变相对应，也提示这些区域对钙释放通道具有重要作用

项目成果

悪性高熱症for professional anesthesiologist(菊地博達 編)

专业麻醉师的恶性高热（菊地博达主编）

• 发表时间: 2006
• 作者: Takashi Makino;Takashi Makino et al.;Hideto Oyamada;Masaaki Tanaka;Masaaki Tanaka;Masaaki Tanaka et al.;Takuya Kikuchi;Takuya Kikuchi et al.;小山田英人
• 通讯作者: 小山田英人

A novel role for the amino terminus of the ryanodine receptor, a Ca release channel, as the retention signal to the endoplasmic reticulum

兰尼碱受体氨基末端（Ca 释放通道）的新作用，作为内质网的保留信号

• 发表时间: 2005
• 期刊: Showa University Journal of Medical Sciences (印刷中)
• 作者: Takashi Makino;Takashi Makino et al.;Hideto Oyamada;Masaaki Tanaka
• 通讯作者: Masaaki Tanaka

Distribution of retention signals to the endoplasmic reticulum in the type 1 ryanodine receptor (Ca^<2+> release channel)

1型兰尼定受体内质网滞留信号的分布（Ca^2释放通道）

• 发表时间: 2007
• 期刊: Showa University Journal of Medical Sciences (印刷中)
• 作者: Takashi Makino

Construction and Expression of Ryanodine Receptor Serial deletion Clones in Chinese Hamster Ovary Cells.

中国仓鼠卵巢细胞中兰尼定受体序列缺失克隆的构建和表达。

• 发表时间: 2003
• 期刊: Showa University Journal of Medical Sciences 15
• 作者: Takashi Makino;Takashi Makino et al.;Hideto Oyamada;Masaaki Tanaka;Masaaki Tanaka;Masaaki Tanaka et al.;Takuya Kikuchi
• 通讯作者: Takuya Kikuchi

MALIGNANT HYPERTHERMIA for Professional Anesthesiologists (ed by Hirosatsu Kikuchi, MD and Ph.D)

专业麻醉师的恶性高热（由医学博士和博士 Hirosatsu Kikuchi 编辑）

• 发表时间: 2006
• 期刊: Kokuseido Co LTD
• 作者: Takashi Makino;Takashi Makino et al.;Hideto Oyamada
• 通讯作者: Hideto Oyamada