| Normal and scrapie-associated forms of prion protein differ in their sensitivities to phospholipase and proteases in intact neuroblastoma cells. | |
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MedLine Citation:
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PMID: 1968104 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Previous studies have indicated that scrapie infection results in the accumulation of a proteinase K-resistant form of an endogenous brain protein generally referred to as prion protein (PrP). The molecular nature of the scrapie-associated modification of PrP accounting for proteinase K resistance is not known. As an approach to understanding the cellular events associated with the PrP modification in brain tissue, we sought to identify proteinase K-resistant PrP (PrP-res) in scrapie-infected neuroblastoma cells in vitro and to compare properties of PrP-res with those of its normal proteinase K-sensitive homolog, PrP-sen. PrP-res was detected by immunoblot in scrapie-infected but not uninfected neuroblastoma clones. Densitometry of immunoblots indicated that there was two- to threefold more PrP-res than PrP-sen in one infected clone. Metabolic labeling and membrane immunofluorescence experiments indicated that PrP-sen was located on the cell surface and could be removed from intact cells by phosphatidylinositol-specific phospholipase C and proteases. In contrast, PrP-res was not removed after reaction with these enzymes. Thus, either the scrapie-associated PrP-res was not on the cell surface or it was there in a form that is resistant to these hydrolytic enzymes. Attempts to detect intracellular PrP-res by immunofluorescent staining of fixed and permeabilized cells revealed that PrP was present in discrete perinuclear Golgi-like structures. However, the staining pattern was similar in both scrapie-infected and uninfected clones, and thus the intracellular staining may have represented only PrP-sen. Analysis of scrapie infectivity in cells treated with extracellular phospholipase, proteinase K, and trypsin indicated that, like PrP-res, the scrapie agent was not removed from the infected cells by any of these enzymes. |
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Authors:
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B Caughey; K Neary; R Buller; D Ernst; L L Perry; B Chesebro; R E Race |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Journal of virology Volume: 64 ISSN: 0022-538X ISO Abbreviation: J. Virol. Publication Date: 1990 Mar |
Date Detail:
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Created Date: 1990-03-26 Completed Date: 1990-03-26 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 0113724 Medline TA: J Virol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 1093-101 Citation Subset: IM |
Affiliation:
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Laboratory of Persistent Viral Diseases, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Endopeptidase K Flow Cytometry Fluorescent Antibody Technique Immunoblotting Neuroblastoma PrPSc Proteins Prions / metabolism* Serine Endopeptidases / metabolism* Trypsin / metabolism* Type C Phospholipases / metabolism* Viral Proteins / analysis, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/PrPSc Proteins; 0/Prions; 0/Viral Proteins; EC 3.1.4.-/Type C Phospholipases; EC 3.4.21.-/Serine Endopeptidases; EC 3.4.21.4/Trypsin; EC 3.4.21.64/Endopeptidase K |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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