| Project/Area Number |
06672173
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Biological pharmacy
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| Research Institution | Tokyo Medical & Dental University |
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Principal Investigator |
ISHIDATE Kozo Tokyo Medical & Dental University, Medical Research Institute, Associate Professor., 難治疾患研究所, 助教授 (40014287)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUO Ritsuko Tokyo Medical & Dental University, Medical Research Institute, Research Associat, 難治疾患研究所, 助手 (00126260)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | choline (ethanolamine) kinase / phosphatidylcholine / phosphatidylethanolamine / cDNA cloning / phosphocholine / phosphoethanolamine / ホスホコリン / ホスホエタノールアミン / コリンキナーゼ |
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| Research Abstract |
Choline (ethanolamine) kinase, a key enzyme of phosphatidylcholine (phosphatidylethanolamine) biosynthesis, has been shown to exist in animal cells as several isozymes. Its cDNA cloning has been done for the yeast enzyme, the major from of rat liver enzyme as well as for the human glioblastoma enzyme. Rat kidney enzyme has been purified to homogeneity and its molecular size (42 kDa) is shown to be rather small when compared to either rat liver or human glioblastoma enzyme, which has been reported to have 50 kDa or 50 kDa size, respectively. In this project, we first purified 42 kDa choline (ethanolamine) kinase (CK,EK) and obtained the partial amino acid (AA) sequences of several peptides derived from the purified enzyme by trypsin digestion. Any of the AA sequences thus obtained were not identical to the reported AA sequences of not only other CK (EK) isozymes but also any other proteins. Then, we prepared several mixed oligonucleotide probes to screen the cDNA library from rat kidney, which had been constructed on lambda gt 10. By now, several positive clones have been isolated and the nucleotide sequence of inserted cDNAs has been determined. However, any of these clones have not been shown to have exact rat kidney 42 kDa CK (EK) sequences. The search for the kidney type CK cDNA is still under investigation.
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