| Project/Area Number |
08670932
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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 |
Pediatrics
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| Research Institution | The Tokyo Metropolitan Institute of Medical Science |
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Principal Investigator |
SAKURABA Hitoshi The Tokyo Metropolitan Institute of Medical Science, Department of Clinical Genetics, Researcher, 臨床遺伝学研究部門, 研究員 (60114493)
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| Co-Investigator(Kenkyū-buntansha) |
KASE Ryoichi The Tokyo Metropolitan Institute of Medical Science, Department of Clinical Gene, 臨床遺伝学研究部門, 研究員 (20150203)
SHIMMOTO Michie The Tokyo Metropolitan Institute of Medical Science, Department of Clinical Gene, 臨床遺伝学研究部門, 研究員 (20216237)
ITOH Kohji The Tokyo Metropolitan Institute of Medical Science, Department of Clinical Gene, 臨床遺伝学研究部門, 研究員 (00184656)
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| Project Period (FY) |
1996 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1998: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | alpha-galactosidase / Fabry disease / protective protein / galactosialidosis / GM2 gangliosidosis / GM2 activator / メタノール資化酵母 / X線結晶構造解析 / 遺伝子変異 / アンチセンスオリゴDNA / トランスジェニックマウス / バキュロウイルス / サポシン / 遺伝子解析 / SSCP |
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| Research Abstract |
1. alpha-Galactosidase and Fabry disease
We have expressed recombinant human alpha -galactosidase in Pichia pastoris and determined its crystal structure at 2.6A resolution. The human alpha-galactosidase consists of a catalytic domain, which forms barrel-like structure, and a subdomain including beta-stranded sheets . The structural analysis would facilitate development of therapy for Fabry disease.
2. Protective protein and galactosialidosis
We characterized a defective protective protein gene product with a K453E mutation newly found in a patient with galactosialidosis. Immunocytochemical, expression and metabolic studies revealed that the precursor protective protein was synthesized but it hardly processed to the mature form and degraded in the mutant. Structural model of the mutant protective protein was constructed by replacement of the amino acid residue on the crystal structure of the wild type protective protein precursor reported. The result showed that the K453E mutation would locate at the dimer interface of the protective protein and reduce the hydrogen bond formation in the dimer. The structural change might cause instability of the protective protein dimer.
3. GM2 activator and GM2 gangliosidosis AB variant
We have determined clinical features and biochemical basis of a Japanese patient with GM2 gangliosidosis AB variant, In the patient's cells no mature GM2 activator was detected and the catabolism of GM1 ganglioside was blocked at the level of GM2 ganglioside.
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