NR AWPH
AU Tsukui,K.; Tadokoro,K.; Takata,M.; Yokoyama,T.; Onodera,T.
TI PrPres-like proteins in scrapie-infected hamster plasma similar with the PrPres in brain
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions DIA-57
PT Konferenz-Poster
AB PrPres has hardly been detected in the blood except in leukocytes even during the disease period of animal models, when PrPres is found at high amounts in sensitive tissues. We examined detecting PrPres in scrapie infected hamster plasma (scHaPl) by partial purification and using highly sensitive chemiluminescence immunoblot (IB). Brain homogenate of scrapie-infected hamster (scHaBrh) and scHaPl were obtained at the terminal stage of scrapie (Sc237) infection. The scHaBrh and the scHaPl were digested with PK or PK and PNGase F then purified partially by acidic SDS precipitation. Preparations has analyzed by conventional IB using anti-PrP mAbs and a highly sensitive chemiluminescence reagent (SSWF). Acidic SDS condition was successfully precipitated the PrPres in scHaBrh as well as 3F4-reactive proteins in scHaPl selectively and the SSWF enable the detection of the PrP protein at concentrations as low as in 10-9 g brain equivalent. In combination with an image analyzer, treatment of both scHaBrh and scHaPl with PK and PNGase F resulted in the formation of 19-20kDa 3F4-reactive proteins, indicated similar peptide backbones. Acidic SDS precipitation after the PK treatment of the scHaPl discriminated the scHaPl and mock infected (mc) HaPl but between the 3F4 reactive protein species and the protein species of PrPres in scHaBrh showed extreme Mw similarities. Furthermore, 2DE analysis also showed highly similar patterns between scHaBrh and scHaPl on PrPres and 3F4-reactive protein spots, respectively. In conclusion, the 3F4-reactive proteins in scHaPl are expected to be the PrPres in plasma. From these results, a practical screening test for TSE using plasma is expected to develop on the near future.
AD K. Tsukui, K. Tadokoro: Central Blood Institute, The Japanese Red Cross Society 2-1-67 Tatsumi, Koto-ku, Tokyo135-8521, Japan; M. Takata, T. Yokoyama: Research Center for Prion Diseases, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba-shi, Ibaraki 305-0856, Japan; T. Onodera: School of Agriculture and Life Sciences, Tokyo University, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. E-mail: t141810a@s4.dion.ne.jp
SP englisch
PO Italien