Mutant D178N prion protein converts spontaneously in RT-QuIC assay.
- Author:
Chen GAO
1
;
Ke REN
;
Long-Zhu LI
;
Hui-Ying JIANG
;
Cao CHEN
;
Jin ZHANG
;
Jun HAN
;
Xiao-Ping DONG
Author Information
- Publication Type:Journal Article
- MeSH: Creutzfeldt-Jakob Syndrome; etiology; Humans; Mutant Proteins; genetics; Nucleic Acid Amplification Techniques; methods; Prions; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(5):370-373
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the conversion of mutant D178N prion protein in RT-QuIC assay.
METHODSThe D178N mutant prion PRNP was generated by the method of single site mutation. The mutant PRNP gene was inserted into plasmids of pET24. The full and N-truncated recombinant human prion proteins were expressed and purified. The fibril formations of these proteins were real-time monitored by the method of RT-QuIC. The ability to resist proteinase K (PK) of these fibrils was analyzed.
RESULTSWe succeed to construct human PrP-D178N plamids. The N-truncated human prion protein with D178N (PrP90-231-D178N) can convert spontaneously in RT-QuIC, while full length of human prion D178N protein (PrP23-231-D178N) fails to convert spontaneously. The spontaneously generated fibril has been domenstrated it is partily PK-resistant.
CONCLUSIONThe N-terminal of prion protein (23-90) plays an important role for the D178N mutant protein spontaneously conversion, which provide the clues for study the pathogenesis of genetic CJD.
