De novo generation of infectious prions with bacterially expressed recombinant prion protein. Issue 12 (22nd August 2013)

• Record Type: Journal Article
• Authors: Zhang, Zhihong; Zhang, Yi; Wang, Fei; Wang, Xinhe; Xu, Yuanyuan; Yang, Huaiyi; Yu, Guohua; Yuan, Chonggang; Ma, Jiyan
• Is Part Of: FASEB journal. Volume 27:Issue 12(2013)
• Page Start: 4768
• Page End: 4775
• Publication Date: 2013-08-22
• Subjects: sPMCA -- recombinant PrP -- bioassay; Biology -- Periodicals; Biology, Experimental -- Periodicals; 570
• DOI: 10.1096/fj.13-233965
• Languages: English
• ISSNs: 0892-6638

Abstract

Abstract : The prion hypothesis is strongly supported by the fact that prion infectivity and the pathogenic conformer of prion protein (PrP) are simultaneously propagated in vitro by the serial protein misfolding cyclic amplification (sPMCA). However, due to sPMCA's enormous amplification power, whether an infectious prion can be formed de novo with bacterially expressed recombinant PrP (rPrP) remains to be satisfactorily resolved. To address this question, we performed unseeded sPMCA with rPrP in a laboratory that has never been exposed to any native prions. Two types of proteinase K (PK)‐resistant and self‐perpetuating recombinant PrP conformers (rPrP‐res) with PK‐resistant cores of 17 or 14 kDa were generated. A bioassay revealed that rPrP‐res 17kDa was highly infectious, causing prion disease in wild‐type mice with an average survival time of about 172 d. In contrast, rPrP‐res 14kDa completely failed to induce any disease. Our findings reveal that sPMCA is sufficient to initiate various self‐perpetuating PK‐resistant rPrP conformers, but not all of them possess in vivo infectivity. Moreover, generating an infectious prion in a prion‐free environment establishes that an infectious prion can be formed de novo with bacterially expressed rPrP.—Zhang, Z., Zhang, Y., Wang, F., Wang, X., Xu, Y., Yang, H., Yu, G., Yuan, C., Ma, J., De novo generation of infectious prions with bacterially expressed recombinant prion protein. FASEB J. 27, 4768–4775 (2013). www.fasebj.org