| Project/Area Number |
16591779
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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 |
Ophthalmology
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| Research Institution | National Hospital Organization Tokyo Medical Center |
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Principal Investigator |
IWATA Takeshi National Hospital Organization Tokyo Medical Center, National Institute of Sensory Organs, Laboratory Head, 視覚研究部門, 室長 (90374157)
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| Co-Investigator(Kenkyū-buntansha) |
NISHIMURA Toshihide Tokyo Medical College, Clinical Proteome Research Center, Professor, 臨床プロテオーム研究センター, 教授 (40366092)
西村 俊秀 東京医科大学, 臨床プロテオーム研究センター, 教授 (40663092)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | glaucoma / optineurin / Rab8 / retinal ganglion cells / protein interaction / quartz crystal balance / gene mutation / intra ocular pressure / タンパク質相互作用 / 小胞体 / 正常眼圧力緑内障 / アストロサイト |
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| Research Abstract |
Glaucoma is one of the leading causes of blindness affecting 70 million people world wide. Glaucoma is often associated with elevated intraocular pressure (IOP) which leads to specific cell death of the retinal ganglion cells (RGC). However, patients with normal tension glaucoma (NTG), a subtype of primary open angle glaucoma (POAG), are affected without the elevation of IOP. In Japan, more than 98% of patients affected by POAG are diagnosed as NTG. Molecular pathways leading to the pathology of the disease are still unclear mainly due to the lack of information of proteins responsible for the disease.
We have focused on optineurin (OPTN), which was identified as the first gene for NTG. Do to the location of the mutation E50K, which is responsible for severe NTG, we decided to analyze the protein-protein interaction between OPTN and Rab8. Interaction was measured by Quartz-Crystal Microbalance (QCM) and GST-pull down assay. All OPTN mutants and Rab8 where expressed in bacteria and purified by affinity chromatography and gel filtration. Each mutant was added to the reaction chamber where wild type Rab8 was fixed on QCM sensor chip. Only E50K showed no interaction with Rab8. This result was later confirmed by the pull down assay.
Rab8 belongs to the family of Rab8 GTP-binding proteins which is functionally known to regulate protein trafficking in the cell. Disruption of interaction between OPTN-Rab8 may explain the cause of specific cell death of retinal ganglion cells (RGC). We are currently over expressing the mutant E50K to observe organelle change in RGC.
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