1999 Fiscal Year Final Research Report Summary
The role of glycocylphosphatidylinositol-anchor proteins in cell proliferation and differenciation.
| Project/Area Number |
10672080
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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 |
Biological pharmacy
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| Research Institution | Mukogawa Women's University |
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Principal Investigator |
NAKABAYASHI Toshikatsu Mukogawa Women's University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (30128665)
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| Project Period (FY) |
1998 – 1999
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| Keywords | TBR 31-2 cells / proliferation and differentiation / osteoblast / adipocyte / differentiation factor / differentiation marker / GPI-anchor proteins |
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| Research Abstract |
(1) Differentiation capacity of TBR 31-2 cells
To study the differentiation capacity of TBR 31-2 cells, the m-RNA expression of differentiation factor, osteoblast core binding factor 1and differentiation phenotypes such as alkaline phosphatase, type I collagen, osteocalcin, osteopontin, PTH receptor and vitamin D receptor were examined by reverse transcription-polymerase chain reaction. Expression of these proteins was only observed in the stage of cell differentiation. In addition, the increase of m-RNA expression level in differentiation factor, peroxisome proliferator-activated receptor γ and differentiation markers such as lipoprotein lipase and adipsin were also observed. These observations suggest that TBR 31-2 cells show bipotential characters, differentiation toward adipocytes and osteoblasts.
(2) Purification and identification of GPI-anchor proteins in TBR 31-2 cells
After biotinylation of peripheral proteins present on the plasma membranes of TBR 31-2 cells, GPI-anchor proteins were released by the treatment of PI-PLC. Solubilized GPI-anchor proteins were separated by SDS-PAGE, blotted to nitrocellulose membrane, and then analyzed chemiluminescence using peroxidase-conjugated streptoavidin. Four proteins having molecular weights of 120, 112, 96 and 60kDa were specifically solubilized by the action of PI-PLC.
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