Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126

Y. Gu, Y. Jing, A. Kumar, Y. Sharma, H. Fujioka, N. Singh

Research output: Contribution to journalArticle

Abstract

Prion diseases or transmissible spongiform encephalopathies, are neurodegenerative disorders that are genetic, sporadic, or infectious. The pathogenetic event common to all prion disorders is the conformational transformation of the cellular prion protein (PrPC) to the scrapie form (PrPSc), that deposits in the brain parenchyma and induces neuronal death. Infectious prion disorders are caused by exogenously introduced PrPSc that acts as a template in the conversion of endogenous PrPC to nascent PrPSc, and subsequently the process becomes autocatalytic. To understand the process of cellular uptake of PrPSc and its mechanism of cellular toxicity, previous studies have used a PrP fragment spanning residues 106-126 (PrPTx) that is toxic to primary neurons in culture, and mimics PrPSc in its biophysical properties [9,11,14]. Several possible mechanisms of cell death by PrPTx have been proposed [2,3,10,11,18], but the existing data are unclear. To identify the biochemical pathways of neurotoxicity by this fragment, we have isolated mutant neuroblastoma and NT-2 cells that are resistant to toxicity by PrPTx. We show that these cells bind and internalize PrPTx in a temperature dependent fashion, and the peptide accumulates in intracellular compartments, probably lysosomes, where it has an unusually long half-life. The PrPTx-resistant phenotype of the cells reported in this study could result from aberrant binding or internalization of the peptide, or due to an abnormality in the downstream pathway(s) of neuronal toxicity. The PrPTx-resistant cells are therefore a useful tool for evaluating the cellular and biochemical pathways that lead to cell death by this peptide, and will provide insight into the mechanism(s) of neurotoxicity by PrPSc.

Original languageEnglish (US)
Pages (from-to)169-180
Number of pages12
JournalJournal of Alzheimer's Disease
Volume3
Issue number2
StatePublished - 2001
Externally publishedYes

Fingerprint

Scrapie
Prion Diseases
Prions
Peptides
Cell Death
Poisons
Lysosomes
Neuroblastoma
Neurodegenerative Diseases
Half-Life
peptide 106
prion protein (106-126)
Phenotype
Neurons
Temperature
Brain

Keywords

  • Prion
  • Resistant cells
  • Toxic peptide

ASJC Scopus subject areas

  • Neuropsychology and Physiological Psychology

Cite this

Gu, Y., Jing, Y., Kumar, A., Sharma, Y., Fujioka, H., & Singh, N. (2001). Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126. Journal of Alzheimer's Disease, 3(2), 169-180.

Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126. / Gu, Y.; Jing, Y.; Kumar, A.; Sharma, Y.; Fujioka, H.; Singh, N.

In: Journal of Alzheimer's Disease, Vol. 3, No. 2, 2001, p. 169-180.

Research output: Contribution to journalArticle

Gu, Y, Jing, Y, Kumar, A, Sharma, Y, Fujioka, H & Singh, N 2001, 'Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126', Journal of Alzheimer's Disease, vol. 3, no. 2, pp. 169-180.
Gu, Y. ; Jing, Y. ; Kumar, A. ; Sharma, Y. ; Fujioka, H. ; Singh, N. / Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126. In: Journal of Alzheimer's Disease. 2001 ; Vol. 3, No. 2. pp. 169-180.
@article{8a5b78ab062e4f1d8baf2b8fd97a1217,
title = "Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126",
abstract = "Prion diseases or transmissible spongiform encephalopathies, are neurodegenerative disorders that are genetic, sporadic, or infectious. The pathogenetic event common to all prion disorders is the conformational transformation of the cellular prion protein (PrPC) to the scrapie form (PrPSc), that deposits in the brain parenchyma and induces neuronal death. Infectious prion disorders are caused by exogenously introduced PrPSc that acts as a template in the conversion of endogenous PrPC to nascent PrPSc, and subsequently the process becomes autocatalytic. To understand the process of cellular uptake of PrPSc and its mechanism of cellular toxicity, previous studies have used a PrP fragment spanning residues 106-126 (PrPTx) that is toxic to primary neurons in culture, and mimics PrPSc in its biophysical properties [9,11,14]. Several possible mechanisms of cell death by PrPTx have been proposed [2,3,10,11,18], but the existing data are unclear. To identify the biochemical pathways of neurotoxicity by this fragment, we have isolated mutant neuroblastoma and NT-2 cells that are resistant to toxicity by PrPTx. We show that these cells bind and internalize PrPTx in a temperature dependent fashion, and the peptide accumulates in intracellular compartments, probably lysosomes, where it has an unusually long half-life. The PrPTx-resistant phenotype of the cells reported in this study could result from aberrant binding or internalization of the peptide, or due to an abnormality in the downstream pathway(s) of neuronal toxicity. The PrPTx-resistant cells are therefore a useful tool for evaluating the cellular and biochemical pathways that lead to cell death by this peptide, and will provide insight into the mechanism(s) of neurotoxicity by PrPSc.",
keywords = "Prion, Resistant cells, Toxic peptide",
author = "Y. Gu and Y. Jing and A. Kumar and Y. Sharma and H. Fujioka and N. Singh",
year = "2001",
language = "English (US)",
volume = "3",
pages = "169--180",
journal = "Journal of Alzheimer's Disease",
issn = "1387-2877",
publisher = "IOS Press",
number = "2",

}

TY - JOUR

T1 - Isolation of human neuronal cells resistant to toxicity by the prion protein peptide 106-126

AU - Gu, Y.

AU - Jing, Y.

AU - Kumar, A.

AU - Sharma, Y.

AU - Fujioka, H.

AU - Singh, N.

PY - 2001

Y1 - 2001

N2 - Prion diseases or transmissible spongiform encephalopathies, are neurodegenerative disorders that are genetic, sporadic, or infectious. The pathogenetic event common to all prion disorders is the conformational transformation of the cellular prion protein (PrPC) to the scrapie form (PrPSc), that deposits in the brain parenchyma and induces neuronal death. Infectious prion disorders are caused by exogenously introduced PrPSc that acts as a template in the conversion of endogenous PrPC to nascent PrPSc, and subsequently the process becomes autocatalytic. To understand the process of cellular uptake of PrPSc and its mechanism of cellular toxicity, previous studies have used a PrP fragment spanning residues 106-126 (PrPTx) that is toxic to primary neurons in culture, and mimics PrPSc in its biophysical properties [9,11,14]. Several possible mechanisms of cell death by PrPTx have been proposed [2,3,10,11,18], but the existing data are unclear. To identify the biochemical pathways of neurotoxicity by this fragment, we have isolated mutant neuroblastoma and NT-2 cells that are resistant to toxicity by PrPTx. We show that these cells bind and internalize PrPTx in a temperature dependent fashion, and the peptide accumulates in intracellular compartments, probably lysosomes, where it has an unusually long half-life. The PrPTx-resistant phenotype of the cells reported in this study could result from aberrant binding or internalization of the peptide, or due to an abnormality in the downstream pathway(s) of neuronal toxicity. The PrPTx-resistant cells are therefore a useful tool for evaluating the cellular and biochemical pathways that lead to cell death by this peptide, and will provide insight into the mechanism(s) of neurotoxicity by PrPSc.

AB - Prion diseases or transmissible spongiform encephalopathies, are neurodegenerative disorders that are genetic, sporadic, or infectious. The pathogenetic event common to all prion disorders is the conformational transformation of the cellular prion protein (PrPC) to the scrapie form (PrPSc), that deposits in the brain parenchyma and induces neuronal death. Infectious prion disorders are caused by exogenously introduced PrPSc that acts as a template in the conversion of endogenous PrPC to nascent PrPSc, and subsequently the process becomes autocatalytic. To understand the process of cellular uptake of PrPSc and its mechanism of cellular toxicity, previous studies have used a PrP fragment spanning residues 106-126 (PrPTx) that is toxic to primary neurons in culture, and mimics PrPSc in its biophysical properties [9,11,14]. Several possible mechanisms of cell death by PrPTx have been proposed [2,3,10,11,18], but the existing data are unclear. To identify the biochemical pathways of neurotoxicity by this fragment, we have isolated mutant neuroblastoma and NT-2 cells that are resistant to toxicity by PrPTx. We show that these cells bind and internalize PrPTx in a temperature dependent fashion, and the peptide accumulates in intracellular compartments, probably lysosomes, where it has an unusually long half-life. The PrPTx-resistant phenotype of the cells reported in this study could result from aberrant binding or internalization of the peptide, or due to an abnormality in the downstream pathway(s) of neuronal toxicity. The PrPTx-resistant cells are therefore a useful tool for evaluating the cellular and biochemical pathways that lead to cell death by this peptide, and will provide insight into the mechanism(s) of neurotoxicity by PrPSc.

KW - Prion

KW - Resistant cells

KW - Toxic peptide

UR - http://www.scopus.com/inward/record.url?scp=0034990259&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034990259&partnerID=8YFLogxK

M3 - Article

VL - 3

SP - 169

EP - 180

JO - Journal of Alzheimer's Disease

JF - Journal of Alzheimer's Disease

SN - 1387-2877

IS - 2

ER -