Abstract
Prion diseases are neurodegenerative disorders associated with the accumulation of an abnormal isoform of the mammalian prion protein (PrP). Fourier transform infrared spectroscopy (FTIR) has previously been used to show that the conformation of aggregated, infectious PrP (PrP Sc) varies between prion strains and these unique conformations may determine strain-specific disease phenotypes. However, the relative amounts of α-helix, β-sheet and other secondary structures have not always been consistent between studies, suggesting that other proteins might be confounding the analysis of PrP Sc secondary structure. We have used FTIR and LC-MS/MS to analyze enriched PrP Sc from mouse and hamster prion strains both before and after the removal of protein contaminants that commonly co-purify with PrP Sc. Our data show that non-PrP proteins do contribute to absorbances that have been associated with α-helical, loop, turn and β-sheet structures attributed to PrP Sc. The major contaminant, the α-helical protein ferritin, absorbs strongly at 1652cm -1 in the FTIR spectrum associated with PrP Sc. However, even the removal of more than 99% of the ferritin from PrP Sc did not completely abolish absorbance at 1652cm -1. Our results show that contaminating proteins alter the FTIR spectrum attributed to PrP Sc and suggest that the α-helical, loop/turn and β-sheet secondary structure that remains following their removal are derived from PrP Sc itself.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3853-3865 |
| Number of pages | 13 |
| Journal | Proteomics |
| Volume | 11 |
| Issue number | 19 |
| DOIs | |
| State | Published - Oct 2011 |
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Keywords
- Animal proteomics
- Apolipoprotein E
- Ferritin
- Infrared spectroscopy
- Prion protein purification
- Prion strains
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry
Cite this
Identification and removal of proteins that co-purify with infectious prion protein improves the analysis of its secondary structure. / Moore, Roger A.; Timmes, Andrew G.; Wilmarth, Phillip; Safronetz, David; Priola, Suzette A.
In: Proteomics, Vol. 11, No. 19, 10.2011, p. 3853-3865.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Identification and removal of proteins that co-purify with infectious prion protein improves the analysis of its secondary structure
AU - Moore, Roger A.
AU - Timmes, Andrew G.
AU - Wilmarth, Phillip
AU - Safronetz, David
AU - Priola, Suzette A.
PY - 2011/10
Y1 - 2011/10
N2 - Prion diseases are neurodegenerative disorders associated with the accumulation of an abnormal isoform of the mammalian prion protein (PrP). Fourier transform infrared spectroscopy (FTIR) has previously been used to show that the conformation of aggregated, infectious PrP (PrP Sc) varies between prion strains and these unique conformations may determine strain-specific disease phenotypes. However, the relative amounts of α-helix, β-sheet and other secondary structures have not always been consistent between studies, suggesting that other proteins might be confounding the analysis of PrP Sc secondary structure. We have used FTIR and LC-MS/MS to analyze enriched PrP Sc from mouse and hamster prion strains both before and after the removal of protein contaminants that commonly co-purify with PrP Sc. Our data show that non-PrP proteins do contribute to absorbances that have been associated with α-helical, loop, turn and β-sheet structures attributed to PrP Sc. The major contaminant, the α-helical protein ferritin, absorbs strongly at 1652cm -1 in the FTIR spectrum associated with PrP Sc. However, even the removal of more than 99% of the ferritin from PrP Sc did not completely abolish absorbance at 1652cm -1. Our results show that contaminating proteins alter the FTIR spectrum attributed to PrP Sc and suggest that the α-helical, loop/turn and β-sheet secondary structure that remains following their removal are derived from PrP Sc itself.
AB - Prion diseases are neurodegenerative disorders associated with the accumulation of an abnormal isoform of the mammalian prion protein (PrP). Fourier transform infrared spectroscopy (FTIR) has previously been used to show that the conformation of aggregated, infectious PrP (PrP Sc) varies between prion strains and these unique conformations may determine strain-specific disease phenotypes. However, the relative amounts of α-helix, β-sheet and other secondary structures have not always been consistent between studies, suggesting that other proteins might be confounding the analysis of PrP Sc secondary structure. We have used FTIR and LC-MS/MS to analyze enriched PrP Sc from mouse and hamster prion strains both before and after the removal of protein contaminants that commonly co-purify with PrP Sc. Our data show that non-PrP proteins do contribute to absorbances that have been associated with α-helical, loop, turn and β-sheet structures attributed to PrP Sc. The major contaminant, the α-helical protein ferritin, absorbs strongly at 1652cm -1 in the FTIR spectrum associated with PrP Sc. However, even the removal of more than 99% of the ferritin from PrP Sc did not completely abolish absorbance at 1652cm -1. Our results show that contaminating proteins alter the FTIR spectrum attributed to PrP Sc and suggest that the α-helical, loop/turn and β-sheet secondary structure that remains following their removal are derived from PrP Sc itself.
KW - Animal proteomics
KW - Apolipoprotein E
KW - Ferritin
KW - Infrared spectroscopy
KW - Prion protein purification
KW - Prion strains
UR - http://www.scopus.com/inward/record.url?scp=80052976271&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=80052976271&partnerID=8YFLogxK
U2 - 10.1002/pmic.201100253
DO - 10.1002/pmic.201100253
M3 - Article
C2 - 21805638
AN - SCOPUS:80052976271
VL - 11
SP - 3853
EP - 3865
JO - Proteomics
JF - Proteomics
SN - 1615-9853
IS - 19
ER -