Investigation of PrPC Metabolism and Function in Live Cells

Cathryn L Haigh, David R Brown

Research output: Contribution to journalArticle

  • 3 Citations

Abstract

Prion protein (PrP)C expression levels and protein localization are known to be affected by factors such as metal ions and oxidative stress. By the development of a green fluorescent protein (GFP)-PrPC fusion protein, the movement of PrP can be followed in real time. Furthermore, alterations in cellular metabolism can be detected while cells are still viable. The internalization response of PrP to 20 μM manganese (Mn) in divalent metal ion-depleted media is used to demonstrate the movement of GFP-tagged proteins in live cells and real tim0e. A live cell microtiter plate assay shows that PrP null cells are less capable of dealing with Mn-induced oxidative stress. In addition, this chapter outlines several complementary techniques for studying live cells and GFP fusion proteins.
LanguageEnglish
Pages21-34
Number of pages14
JournalMethods in Molecular Biology
Volume459
DOIs
StatusPublished - 2008

Fingerprint

Protein C
Green Fluorescent Proteins
Manganese
PrPC Proteins
Oxidative Stress
Metals
Ions
Null Lymphocytes
Proteins
Prion Proteins

Cite this

Investigation of PrPC Metabolism and Function in Live Cells. / Haigh, Cathryn L; Brown, David R.

In: Methods in Molecular Biology, Vol. 459, 2008, p. 21-34.

Research output: Contribution to journalArticle

@article{b9c9f578c9c34b6eb3eae096e6fc12d6,
title = "Investigation of PrPC Metabolism and Function in Live Cells",
abstract = "Prion protein (PrP)C expression levels and protein localization are known to be affected by factors such as metal ions and oxidative stress. By the development of a green fluorescent protein (GFP)-PrPC fusion protein, the movement of PrP can be followed in real time. Furthermore, alterations in cellular metabolism can be detected while cells are still viable. The internalization response of PrP to 20 μM manganese (Mn) in divalent metal ion-depleted media is used to demonstrate the movement of GFP-tagged proteins in live cells and real tim0e. A live cell microtiter plate assay shows that PrP null cells are less capable of dealing with Mn-induced oxidative stress. In addition, this chapter outlines several complementary techniques for studying live cells and GFP fusion proteins.",
author = "Haigh, {Cathryn L} and Brown, {David R}",
year = "2008",
doi = "10.1007/978-1-59745-234-2_2",
language = "English",
volume = "459",
pages = "21--34",
journal = "Methods in Molecular Biology",
issn = "1064-3745",
publisher = "Humana Press",

}

TY - JOUR

T1 - Investigation of PrPC Metabolism and Function in Live Cells

AU - Haigh,Cathryn L

AU - Brown,David R

PY - 2008

Y1 - 2008

N2 - Prion protein (PrP)C expression levels and protein localization are known to be affected by factors such as metal ions and oxidative stress. By the development of a green fluorescent protein (GFP)-PrPC fusion protein, the movement of PrP can be followed in real time. Furthermore, alterations in cellular metabolism can be detected while cells are still viable. The internalization response of PrP to 20 μM manganese (Mn) in divalent metal ion-depleted media is used to demonstrate the movement of GFP-tagged proteins in live cells and real tim0e. A live cell microtiter plate assay shows that PrP null cells are less capable of dealing with Mn-induced oxidative stress. In addition, this chapter outlines several complementary techniques for studying live cells and GFP fusion proteins.

AB - Prion protein (PrP)C expression levels and protein localization are known to be affected by factors such as metal ions and oxidative stress. By the development of a green fluorescent protein (GFP)-PrPC fusion protein, the movement of PrP can be followed in real time. Furthermore, alterations in cellular metabolism can be detected while cells are still viable. The internalization response of PrP to 20 μM manganese (Mn) in divalent metal ion-depleted media is used to demonstrate the movement of GFP-tagged proteins in live cells and real tim0e. A live cell microtiter plate assay shows that PrP null cells are less capable of dealing with Mn-induced oxidative stress. In addition, this chapter outlines several complementary techniques for studying live cells and GFP fusion proteins.

UR - http://www.scopus.com/inward/record.url?scp=48849105384&partnerID=8YFLogxK

UR - http://dx.doi.org/10.1007/978-1-59745-234-2

UR - http://dx.doi.org/10.1007/978-1-59745-234-2_2

U2 - 10.1007/978-1-59745-234-2_2

DO - 10.1007/978-1-59745-234-2_2

M3 - Article

VL - 459

SP - 21

EP - 34

JO - Methods in Molecular Biology

T2 - Methods in Molecular Biology

JF - Methods in Molecular Biology

SN - 1064-3745

ER -