title: De novo generation of infectious prions with bacterially expressed recombinant prion protein.
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.
authors: 2Department of Molecular and Cellular Biochemistry, 1645 Neil Ave., Rm. 457A Hamilton Hall, Ohio State University, Columbus, OH 43210, USA. ma.131@osu.edu.
authors: Zhihong
authors: Z
authors: Zhang
authors: Yi
authors: Y
authors: Zhang
authors: Fei
authors: F
authors: Wang
authors: Xinhe
authors: X
authors: Wang
authors: Yuanyuan
authors: Y
authors: Xu
authors: Huaiyi
authors: H
authors: Yang
authors: Guohua
authors: G
authors: Yu
authors: Chonggang
authors: C
authors: Yuan
authors: Jiyan
authors: J
authors: Ma
mesh_terms: D000818:Animals; D001921:Brain / Q000473:pathology; D051379:Mice; D011328:Prions / Q000737:chemistry* / Q000472:pathogenicity; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D011328:Prions; D011994:Recombinant Proteins
keywords: bioassay; recombinant PrP; sPMCA
references: J Biol Chem. 2005 Mar 25;280(12):11247-58
references: 15618225
references: Nat Med. 2005 Sep;11(9):982-5
references: 16127436
references: Cell. 2005 Apr 22;121(2):195-206
references: 15851027
references: J Biol Chem. 2006 Nov 17;281(46):35245-52
references: 16982620
references: Trends Cell Biol. 1997 Feb;7(2):56-62
references: 17708907
references: Biochemistry. 2007 Jun 12;46(23):7045-53
references: 17503780
references: Methods. 2011 Mar;53(3):208-13
references: 21176786
references: Proc Natl Acad Sci U S A. 2012 May 29;109(22):8546-51
references: 22586108
references: Acta Biochim Biophys Sin (Shanghai). 2013 Jun;45(6):485-93
references: 23583976
references: Nat Rev Mol Cell Biol. 2010 Apr;11(4):301-7
references: 20308987
references: FEBS Lett. 2008 Sep 22;582(21-22):3161-6
references: 18706416
references: J Biol Chem. 2010 May 7;285(19):14083-7
references: 20304915
references: Cell. 2011 Oct 28;147(3):498-508
references: 22036560
references: Proc Natl Acad Sci U S A. 2007 Jun 5;104(23):9741-6
references: 17535913
references: Science. 2004 Jul 30;305(5684):673-6
references: 15286374
references: Science. 2010 Feb 26;327(5969):1132-5
references: 20110469
references: PLoS Pathog. 2009 May;5(5):e1000421
references: 19436715
references: Biochem Soc Symp. 2005;(72):211-22
references: 15649144
references: Biochemistry. 2003 Mar 25;42(11):3295-304
references: 12641461
references: Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):10168-72
references: 10963679
references: Nature. 2001 Jun 14;411(6839):810-3
references: 11459061
references: PLoS Pathog. 2010 Jan 22;6(1):e1000736
references: 20107515
references: Proc Natl Acad Sci U S A. 2007 May 1;104(18):7546-51
references: 17456603
references: J Biol Chem. 2008 Jul 11;283(28):19616-25
references: 18456654
references: Nat Protoc. 2012 Jun 28;7(7):1397-409
references: 22743831
references: J Gen Virol. 2008 Jul;89(Pt 7):1777-1788
references: 18559949
references: J Mol Biol. 2003 Sep 5;332(1):47-57
references: 12946346
references: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13363-83
references: 9811807
references: PLoS Pathog. 2009 Jul;5(7):e1000535
references: 19649330
references: Science. 2006 Jul 7;313(5783):92-4
references: 16825570
references: Science. 2012 Jun 22;336(6088):1511-3
references: 22723400
references: Nature. 2009 Jun 18;459(7249):924-5
references: 19536253
references: PLoS Pathog. 2011 Nov;7(11):e1002370
references: 22114554
references: Acta Neuropathol. 2010 Feb;119(2):177-87
references: 20052481
references: J Virol. 2011 Mar;85(6):2582-8
references: 21228241
references: Biochim Biophys Acta. 2009 Jun;1792(6):555-63
references: 19281844
references: J Biol Chem. 1999 Dec 24;274(52):36859-65
references: 10601237
references: J Mol Biol. 2000 Aug 18;301(3):567-73
references: 10966770
references: PLoS Pathog. 2012;8(4):e1002589
references: 22511864
references: J Exp Med. 2012 May 7;209(5):889-93
references: 22566400
references: J Biol Chem. 2012 Oct 26;287(44):37219-32
references: 22948149
references: J Biol Chem. 2008 Jun 6;283(23):15988-96
references: 18400757
references: Proc Natl Acad Sci U S A. 2012 Jul 10;109(28):E1938-46
references: 22711839
references: J Virol. 2012 Feb;86(3):1874-6
references: 22090130
references: Cell. 2012 May 25;149(5):968-77
references: 22632966
references: Neuron. 2009 Dec 24;64(6):783-90
references: 20064386
references: Biochemistry. 2010 Sep 21;49(37):8169-76
references: 20718504