Out with the old, in with the new? Comparing methods for measuring protein degradation.
Publication
, Journal Article
Yewdell, JW; Lacsina, JR; Rechsteiner, MC; Nicchitta, CV
Published in: Cell Biol Int
May 2011
Protein degradation is a critical factor in controlling cellular protein abundance. Here, we compare classical methods for determining protein degradation rates to a novel GFP (green fluorescent protein) fusion protein based method that assesses the intrinsic stability of cloned cDNA library products by flow cytometry [Yen et al. (2008) Science 322, 918]. While no method is perfect, we conclude that chimeric gene reporter approaches, though powerful, should be applied cautiously, due principally to GFP (or other reporter tag) interference with protein organelle targeting or incorporation into macromolecular assemblies, both of which cause spuriously high degradation rates.
Duke Scholars
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Published In
Cell Biol Int
DOI
EISSN
1095-8355
Publication Date
May 2011
Volume
35
Issue
5
Start / End Page
457 / 462
Location
England
Related Subject Headings
- Recombinant Fusion Proteins
- Proteins
- Humans
- Green Fluorescent Proteins
- Flow Cytometry
- Cloning, Molecular
- Biochemistry & Molecular Biology
- Biochemistry
- Animals
- 3101 Biochemistry and cell biology
Citation
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Chicago
ICMJE
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NLM
Yewdell, J. W., Lacsina, J. R., Rechsteiner, M. C., & Nicchitta, C. V. (2011). Out with the old, in with the new? Comparing methods for measuring protein degradation. Cell Biol Int, 35(5), 457–462. https://doi.org/10.1042/CBI20110055
Yewdell, Jonathan W., Joshua R. Lacsina, Martin C. Rechsteiner, and Christopher V. Nicchitta. “Out with the old, in with the new? Comparing methods for measuring protein degradation.” Cell Biol Int 35, no. 5 (May 2011): 457–62. https://doi.org/10.1042/CBI20110055.
Yewdell JW, Lacsina JR, Rechsteiner MC, Nicchitta CV. Out with the old, in with the new? Comparing methods for measuring protein degradation. Cell Biol Int. 2011 May;35(5):457–62.
Yewdell, Jonathan W., et al. “Out with the old, in with the new? Comparing methods for measuring protein degradation.” Cell Biol Int, vol. 35, no. 5, May 2011, pp. 457–62. Pubmed, doi:10.1042/CBI20110055.
Yewdell JW, Lacsina JR, Rechsteiner MC, Nicchitta CV. Out with the old, in with the new? Comparing methods for measuring protein degradation. Cell Biol Int. 2011 May;35(5):457–462.
Published In
Cell Biol Int
DOI
EISSN
1095-8355
Publication Date
May 2011
Volume
35
Issue
5
Start / End Page
457 / 462
Location
England
Related Subject Headings
- Recombinant Fusion Proteins
- Proteins
- Humans
- Green Fluorescent Proteins
- Flow Cytometry
- Cloning, Molecular
- Biochemistry & Molecular Biology
- Biochemistry
- Animals
- 3101 Biochemistry and cell biology