Propagation of a protease-resistant form of prion protein in long-term cultured human glioblastoma cell line T98G
| Autor: | Jun-ichi Sawada, Takeshi Yamazaki, Yutaka Kikuchi, Ken-ichi Tanamoto, Ayako Sakai, Tomoshi Kakeya, Kosuke Takatori, Haruo Matsuda, Naoto Nakamura |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Gene isoform
biology Prions medicine.drug_class animal diseases Immunoblotting Drug Resistance Intracellular Space Proteinase K Monoclonal antibody Virology nervous system diseases Cell culture Polyclonal antibodies Cell Line Tumor Genotype biology.protein medicine Humans Protein Isoforms Endopeptidase K Antibody Glioblastoma Gene |
| Zdroj: | Journal of General Virology. 85:3449-3457 |
| ISSN: | 1465-2099 0022-1317 |
| DOI: | 10.1099/vir.0.80043-0 |
| Popis: | Human prion diseases, such as Creutzfeldt–Jakob disease (CJD), a lethal, neurodegenerative condition, occur in sporadic, genetic and transmitted forms. CJD is associated with the conversion of normal cellular prion protein (PrPC) into a protease-resistant isoform (PrPres). The mechanism of the conversion has not been studied in human cell cultures, due to the lack of a model system. In this study, such a system has been developed by culturing cell lines. Human glioblastoma cell line T98G had no coding-region mutations of the prion protein gene, which was of the 129 M/V genotype, and expressed endogenous PrPC constitutively. T98G cells produced a form of proteinase K (PK)-resistant prion protein fragment following long-term culture and high passage number; its deglycosylated form was approximately 18 kDa. The PK-treated PrPres was detected by immunoblotting with the mAb 6H4, which recognizes residues 144–152, and a polyclonal anti-C-terminal antibody, but not by the mAb 3F4, which recognizes residues 109–112, or the anti-N-terminal mAb HUC2-13. These results suggest that PrPC was converted into a proteinase-resistant form of PrPres in T98G cells. |
| Databáze: | OpenAIRE |
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