SCREENING FOR THE FUNCTIONAL DOMAINS IN THE RYANODINE RECEPTOR
| Project/Area Number |
15590234
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General pharmacology
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| Research Institution | SHOWA UNIVERSITY |
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Principal Investigator |
OGUCHI Katsuji SHOWA UNIVERSITY, SCHOOL OF MEDICINE DEPARTMENT OF PHARMACOLOGY, MD., PhD., PROFESSOR, 医学部, 教授 (50129821)
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| Co-Investigator(Kenkyū-buntansha) |
OYAMADA Hideto SHOWA UNIVERSITY, SCHOOL OF MEDICINE DEPARTMENT OF PHARMACOLOGY, PhD., RESEACH ASSISTANT, 医学部, 助手 (50266160)
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| Project Period (FY) |
2003 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | 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 |
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| Research Abstract |
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
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Report
(5 results)
Research Products
(11 results)
