| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
α-Synuclein is regarded as one of the key molecules when considering the pathogenic mechanisms of a group of Lewy body diseases (LBD), including Parkinson's disease (PD) and diffuse Lewy body disease, in that, first, some mutations of this protein cause familial PD and, second, it is a main proteineous component of the Lewy bodies. Identification and characterization of molecules interacting with this protein is indispensable for the elucidation of the pathogenic mechanisms of LBD.This project focused on the following two points ; first, identification of new molecules interacting with α-synuclein, and second, detailed analyses of the interaction between synuclein and the known interacting molecules. In regard to the first point, we screened a human fetal brain cDNA library using A53T PD-pathogenic mutant synuclein as a bait and obtained some candidate clones, characterization of which is now under way. In addition, by probing metabolically radioisotope-labeled COS-1 proteins with GST-synucleins, we identified a polypeptide with the apparent molecular weight of 72 kDa. which interacts with GST-α-synuclein but not with GST alone. Concerning the latter point, we mainly concentrated on synphilin, which was reported during the period of our project as a protein interacting with α-synuclein. In contradiction to the previously reported results, our results indicate that, as compared with the interaction of an N- and C-terminally truncated synphilin with synuclein, that of full-length wild type one is very week if any, suggesting that presence of either of the N-terminal or C-terminal region acts inhibitory on the interaction.
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