| Project/Area Number |
12680691
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Cell biology
|
|---|
| Research Institution | Mie University |
|---|
Principal Investigator |
OKAGAKI Tsuyoshi Mie University, Department of Bioresource, assistant professor, 生物資源学部, 助教授 (80185412)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KOHAMA Kazuhiro Gunma University, Department of Medicine, professor, 医学部, 教授 (30101116)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | vascular cell / smooth muscle cell / myosin / myosin filament |
|---|
| Research Abstract |
1. Dedifferentiation of smooth muscle cell is key event for induction of atherosclerosis. We isolated and characterized a protein with 38kD that stabilize myosin filament from chicken smooth muscle. From deduced amino acid sequence of cDNA, it was homologue of human p32. This protein has been identified as a mitochondrial protein in some types of cell. Upon incorporation of the protein into mitochondria, N-terminal 70 amino acid residue are deleted by intrinsic protease. We found that the protein is distributed in cultured smooth muscle cell depending on the state of differentiation. This means that the function of the protein is changed upon dedifferentiation. To understand the N-terminal signal sequence on the change of localization, we tried to obtain full length cDNA of the protein. We made cDNA library of chicken gizzard and screened the cDNA, but could not obtained the full length cDNA. The main reason was that the nucleotide coding the signal sequence is quite GC-rich, so that cDNA synthesis was not efficient. Next we obtained pre-made cDNA library and also screened the full length cDNA. Since we could not obtained the full length cDNA, we made cDNA library of HeLa, now we are still doing effort to obtain the full length cDNA
2. To understand molecular mechanism of atherosclerosis, we tried to make model system by means of vascular smooth muscle cell line ACO1. The expression level of smooth muscle specific protein such as α-actin, high molecular weight caldesmon, and desmin was increased upon serum deprivation. By addition of PDGF, expression level of these protein was remarkably reduced. By observation of the localization of myofibrillar protein, cytoskeletal structure was clearly diminished in the presence of PDGF. Thus we could control the level of differentiation by changing the culture condition in vitro.
|
