| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Outline of Final Research Achievements |
To find cellular components affecting the conformation of the cellular prion protein (PrPC), we constructed an assay system measuring trypsin susceptibility of a recombinant cellular prion protein (rPrP). Consequently, we purified heat shock protein 90 (hsp90) from mouse neuroblastoma Neuro-2a cells. Hsp90 is a 90-kDa chaperone protein abundantly expressed in the cytosol, nuclear and extracellular space. In the in vitro assay, 1) Hsp90 unfolded the central region of rPrP, 2) the Hsp90 C-terminal chaperon domain is necessary for the unfolding activity; 3) any nucleotides such as ATP were not necessary for the unfolding activity; 3) Hsp90, however, unfolded the copper-loaded rPrP, which is more resistant to trypsin digestion, in the presence of ATP or ADP, but not AMP. In addition, we obtained novel Hsp90 C-terminal chaperon domain inhibitors, cisplatin derivatives, and confirmed that these inhibitors suppressed the prion formation in prion-infected neuroblastoma cells.
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