| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
We have investigated to elucidate functional of small GTPase rad, which is highly expressed in skeletal muscle of diabetic patients. First we tried to identify rad interacting proteins. We produced and purified GST-tagged rad. Then, we tried the affinity column chromatography uising the reombinant rad. However, we could not identify such proteins.Then, we tried the ligand overlay method. We separated proteins in SDS-PAGE and tranfered tyo nitrocelluloce filter. Then, we incubated the membrane with 32P-rad. After washeng it, the autoradiography was performed but we cannot detect rad interacting proteins. At the moment, since we succeeded in producing with monoclonal antibody agains rad, we are trying coimmmunoprecipitation experiments with the antibody. Furthermore, we are trying cross-linking experiments with 125I-labeled rad and skeletal muscle extract. Then, we detected several rad interacting protein candidates. We are currently purifying them.
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