Details
| Original language | English |
|---|---|
| Title of host publication | Plant Membrane Proteomics |
| Publisher | Humana Press |
| Pages | 163-183 |
| Number of pages | 21 |
| Publication status | Published - 31 Oct 2017 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1696 |
| ISSN (Print) | 1064-3745 |
Abstract
Containing plastids and vacuoles in addition to those organelles also found in other (heterotrophic) cells, the plant cell displays an extraordinary level of compartmentalization, largely obtained by the utilization of membranes. These membranes not only confine reaction spaces but must also facilitate cross-talk between organelles and other cell compartments. They also host important components of the plant energy metabolism, i.e., the electron transport chains of mitochondria and chloroplasts. Characterization of the proteomes of these membranes requires isolation of pure and intact organelles from plant tissues followed by subsequent purification of their respective membranes. Membrane fractions are then amenable for further analyses using gel electrophoresis procedures or gel-free proteomic approaches. Here, we describe the preparation of intact mitochondria from Arabidopsis thaliana cell-culture, the isolation of outer and inner mitochondrial membranes and downstream proteomic applications for analyzing their membrane protein content.
Keywords
- BN-PAGE, Isolation, Mass spectrometry, Mitochondria, Subfractionation, Trypsin digestion
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Genetics
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Plant Membrane Proteomics. Humana Press, 2017. p. 163-183 (Methods in Molecular Biology; Vol. 1696).
Research output: Chapter in book/report/conference proceeding › Contribution to book/anthology › Research › peer review
}
TY - CHAP
T1 - Sample preparation for analysis of the plant mitochondrial membrane proteome
AU - Schikowsky, Christine
AU - Thal, Beate
AU - Braun, Hans Peter
AU - Eubel, Holger
PY - 2017/10/31
Y1 - 2017/10/31
N2 - Containing plastids and vacuoles in addition to those organelles also found in other (heterotrophic) cells, the plant cell displays an extraordinary level of compartmentalization, largely obtained by the utilization of membranes. These membranes not only confine reaction spaces but must also facilitate cross-talk between organelles and other cell compartments. They also host important components of the plant energy metabolism, i.e., the electron transport chains of mitochondria and chloroplasts. Characterization of the proteomes of these membranes requires isolation of pure and intact organelles from plant tissues followed by subsequent purification of their respective membranes. Membrane fractions are then amenable for further analyses using gel electrophoresis procedures or gel-free proteomic approaches. Here, we describe the preparation of intact mitochondria from Arabidopsis thaliana cell-culture, the isolation of outer and inner mitochondrial membranes and downstream proteomic applications for analyzing their membrane protein content.
AB - Containing plastids and vacuoles in addition to those organelles also found in other (heterotrophic) cells, the plant cell displays an extraordinary level of compartmentalization, largely obtained by the utilization of membranes. These membranes not only confine reaction spaces but must also facilitate cross-talk between organelles and other cell compartments. They also host important components of the plant energy metabolism, i.e., the electron transport chains of mitochondria and chloroplasts. Characterization of the proteomes of these membranes requires isolation of pure and intact organelles from plant tissues followed by subsequent purification of their respective membranes. Membrane fractions are then amenable for further analyses using gel electrophoresis procedures or gel-free proteomic approaches. Here, we describe the preparation of intact mitochondria from Arabidopsis thaliana cell-culture, the isolation of outer and inner mitochondrial membranes and downstream proteomic applications for analyzing their membrane protein content.
KW - BN-PAGE
KW - Isolation
KW - Mass spectrometry
KW - Mitochondria
KW - Subfractionation
KW - Trypsin digestion
UR - http://www.scopus.com/inward/record.url?scp=85032625009&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7411-5_11
DO - 10.1007/978-1-4939-7411-5_11
M3 - Contribution to book/anthology
C2 - 29086403
AN - SCOPUS:85032625009
T3 - Methods in Molecular Biology
SP - 163
EP - 183
BT - Plant Membrane Proteomics
PB - Humana Press
ER -