2000 Fiscal Year Final Research Report Summary

Functional expression of small GTPase Rad, highly expressed in skeletal muscle in diabetic patients

Research Project

Project/Area Number	11680629
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	Functional biochemistry
Research Institution	KYOTO UNIVERSITY
Principal Investigator	HISANORI Horinow Graduate school of Medicin, Kyoto University, Instructor, 医学研究科, 助手 (90291426)
Project Period (FY)	1999 – 2000
Keywords	Rad / GTPase / diabetes mellitus / skeletal muscle
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.

(4 results)