| Project/Area Number |
14571413
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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 |
Orthopaedic surgery
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| Research Institution | National Cardiovascular Center Research Institute |
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Principal Investigator |
HINO Jun National Cardiovascular Center Research Institute, Head of Biochemistry, 生化学部, 室長 (40260351)
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| Co-Investigator(Kenkyū-buntansha) |
KAIYA Hiroyuki National Cardiovascular Center Research Institute, Staff of Biochemistry, 生化学部, 室員 (40300975)
KANGAWA Kenji National Cardiovascular Center Research Institute, Director of Biochemistry, 生化学部, 部長 (00112417)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | bone metabolism / biologically active peptide |
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| Research Abstract |
To discover novel biologically active peptides that regulate the bone metabolism, I performed screening using in vitro systematic bioassay for a Ca ion mobilization and cAMP elevating in osteoblastic cells. When I started this project, the paper regarding the peptides that regulate the bone metabolism from rat stomach had been published. In that paper, they discovered the evidence that there is a novel peptide in the stomach. I used the extract of stomach as starting material for screening. Fifty stomachs were collected and extracted in 1M acetic acid solution. The extract was separated by follwing column chromatography ; C-18 Sep-Pak column, ion-exchange column and gel filtration. I found several active fractions of gel filtration chromatography. First I used Ca ion mobilization as a bioactive marker in the assay. I succeeded in purification and isolation of these active substances and determined the amino acid sequences. The analyses showed that the active substances are neuromedin C
… More
(NMC), bradykinin and T-kinin. I first demonstrated in this project that NMC might act as bone metabolism modulator and T-kinin, which was thought to be specific molecular form in plasma, is also present in stomach. Next, some fractions that have cAMP elevating activity were isolated and carried out sequence analyses. The analyses showed that all amino acid sequences of active substances are identical to VIP (vasoactive intestinal polypeptide). But one of them contained some amino acids, which were not identical to VIP.
These Results suggested there are some post-translational modification of amino acid or splicing variants of VIP that have unidentified, although I need to perform further precise analysis. I am doing examination of physiological and biological functions of these four biologically active peptides using in vitiro and in vivo assay system I developed. Furthermore, I am planing to do this project to discover novel biologically active peptides that regulate or control bone metabolism. Less
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