Mutant D178N prion protein converts spontaneously in RT-QuIC assay / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To study the conversion of mutant D178N prion protein in RT-QuIC assay.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>We 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.</p><p><b>CONCLUSION</b>The 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.</p>