Details
| Original language | English |
|---|---|
| Pages (from-to) | 583-594 |
| Number of pages | 12 |
| Journal | Plant Journal |
| Volume | 52 |
| Issue number | 3 |
| Publication status | Published - Nov 2007 |
| Externally published | Yes |
Abstract
Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability of free-flow electrophoresis to assist conventional, centrifugation-based techniques in the preparation of plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy and differential gel electrophoresis. Results indicated a significant increase in purity of the mitochondrial samples, highlighted specific contaminants previously reported as mitochondrial proteins, and also pointed to new means for separating plastids and peroxisomes from mitochondria in plant organellar extracts by exploiting differences in surface charge. This approach has the potential to allow a deeper and more comprehensive investigation of the Arabidopsis organellar proteomes, by providing a second dimension of separation based on surface charge in addition to conventional centrifugation purification protocols relying on size and density.
Keywords
- Centrifugation, Free-flow electrophoresis, Organellar contamination, Plant mitochondria, Proteomics, Sample purity
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
- Agricultural and Biological Sciences(all)
- Plant Science
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: Plant Journal, Vol. 52, No. 3, 11.2007, p. 583-594.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Free-flow electrophoresis for purification of plant mitochondria by surface charge
AU - Eubel, Holger
AU - Lee, Chun Pong
AU - Kuo, John
AU - Meyer, Etienne H.
AU - Taylor, Nicolas L.
AU - Millar, A. Harvey
N1 - Copyright: Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2007/11
Y1 - 2007/11
N2 - Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability of free-flow electrophoresis to assist conventional, centrifugation-based techniques in the preparation of plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy and differential gel electrophoresis. Results indicated a significant increase in purity of the mitochondrial samples, highlighted specific contaminants previously reported as mitochondrial proteins, and also pointed to new means for separating plastids and peroxisomes from mitochondria in plant organellar extracts by exploiting differences in surface charge. This approach has the potential to allow a deeper and more comprehensive investigation of the Arabidopsis organellar proteomes, by providing a second dimension of separation based on surface charge in addition to conventional centrifugation purification protocols relying on size and density.
AB - Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability of free-flow electrophoresis to assist conventional, centrifugation-based techniques in the preparation of plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy and differential gel electrophoresis. Results indicated a significant increase in purity of the mitochondrial samples, highlighted specific contaminants previously reported as mitochondrial proteins, and also pointed to new means for separating plastids and peroxisomes from mitochondria in plant organellar extracts by exploiting differences in surface charge. This approach has the potential to allow a deeper and more comprehensive investigation of the Arabidopsis organellar proteomes, by providing a second dimension of separation based on surface charge in addition to conventional centrifugation purification protocols relying on size and density.
KW - Centrifugation
KW - Free-flow electrophoresis
KW - Organellar contamination
KW - Plant mitochondria
KW - Proteomics
KW - Sample purity
UR - http://www.scopus.com/inward/record.url?scp=35448934538&partnerID=8YFLogxK
U2 - 10.1111/j.1365-313X.2007.03253.x
DO - 10.1111/j.1365-313X.2007.03253.x
M3 - Article
C2 - 17727614
AN - SCOPUS:35448934538
VL - 52
SP - 583
EP - 594
JO - Plant Journal
JF - Plant Journal
SN - 0960-7412
IS - 3
ER -