Expression of cytosolic PrP and analysis of its cytotoxic activities.
- Author:
Xin WANG
1
;
Chen-fang DONG
;
Qi SHI
;
Song SHI
;
Gui-rong WANG
;
Yan-jun LEI
;
Run AN
;
Kun XU
;
Hui-ying JIANG
;
Jun HAN
;
Yun-jun ZHAO
;
Xiao-ping DONG
Author Information
1. College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China. wangxin82-82@163.com
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Cell Survival;
Cytosol;
chemistry;
Endopeptidase K;
pharmacology;
Humans;
Prions;
genetics;
physiology;
Transfection
- From:
Chinese Journal of Virology
2008;24(4):277-281
- CountryChina
- Language:Chinese
-
Abstract:
In order to study the physicochemical characteristics of cytosolic PrP (CytoPrP) and evaluate its possible influence on cell viability, a recombinant plasmid expressing human CytoPrP eukaryoticly was constructed and transfected into human neuroblastoma cell line SH-SY5Y transiently. Proteinase-resistant activities of CytoPrP were evaluated by a proteinase K (PK) digestion and cytotoxic effects of CytoPrP were tested by MTT assay and Trypan Blue cell-counting. The presence of CytoPrP in cytoplasm after transfection was controlled by the presence of protease inhibitor. Compared with wild-type PrP, CytoPrP possessed relatively stronger PK-resistant activities. Obvious cytotoxic effects were observed in the cells after inducement of CytoPrP in cytoplasm by protease inhibitor, showing a dose-dependent manner. The results provide useful scientific evidences for further studies of potential role of CytoPrP in pathological mechanism of prion disease.
