原创力文档relationships between prpsc stability and incubation time for united states scrapie isolates in a natural host system朊蛋白之间的关系稳定和孵化时间美国痒病隔离在一个自然的主机系统.pdfVIP
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relationships between prpsc stability and incubation time for united states scrapie isolates in a natural host system朊蛋白之间的关系稳定和孵化时间美国痒病隔离在一个自然的主机系统
Relationships between PrPSc Stability and Incubation
Time for United States Scrapie Isolates in a Natural Host
System
Catherine E. Vrentas, Justin J. Greenlee, Trudy L. Tatum, Eric M. Nicholson*
National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, Iowa, United States of America
Abstract
Transmissible spongiform encephalopathies (TSEs), including scrapie in sheep (Ovis aries), are fatal neurodegenerative
diseases caused by the misfolding of the cellular prion protein (PrPC ˆ Sc
) into a a-rich conformer (PrP ) that accumulates into
higher-order structures in the brain and other tissues. Distinct strains of TSEs exist, characterized by different pathologic
profiles upon passage into rodents and representing distinct conformations of PrPSc. One biochemical method of
distinguishing strains is the stability of PrPSc as determined by unfolding in guanidine hydrochloride (GdnHCl), which is
tightly and positively correlated with the incubation time of disease upon passage into mice. Here, we utilize a rapid,
protease-free version of the stability assay to characterize naturally occurring scrapie samples, including a fast-acting scrapie
inoculum for which incubation time is highly dependent on the amino acid at codon 136 of the prion protein. We utilize the
stability methodology to identify the presence of two distinct isolates in the inoculum, and compare isolate properties to
those of a host-stabilized reference scrapie isolate (NADC 13-7) in order to assess the stability/incubation time correlation in
a natural host system. We demonstrate the utility of the stability methodology in characterizing TSE isol
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