Details
| Original language | English |
|---|---|
| Pages (from-to) | 640-646 |
| Number of pages | 7 |
| Journal | ELECTROPHORESIS |
| Volume | 23 |
| Issue number | 4 |
| Publication status | Published - 20 Feb 2002 |
Abstract
We report a subdivision of the mitochondrial proteome into defined sets of proteins, which is based on the combination of three different gel electrophoresis procedures. First, Blue-native polyacrylamide gel electrophoresis is employed to separate mitochondrial protein complexes. The protein complexes are electroeluted and completely detached from Coomasssie blue. Subsequently the subunits of the protein complexes are separated by isoelectric focusing and finally by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The resolution capacity of the procedure is demonstrated for the ATP synthase complex, the cytochrome c reductase complex and the preprotein translocase of the outer mitochondrial membrane (the TOM complex). The method allows the separation of isoforms of subunits forming part of protein complexes, whose occurrence seems to be rather a rule than an exception in higher eukaryotes. Furthermore, extremely hydrophobic proteins are detectable on the gels.
Keywords
- Arabidopsis thaliana, Plant mitochondria, Three-dimensional gel electrophoresis
ASJC Scopus subject areas
- Chemistry(all)
- Analytical Chemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Clinical Biochemistry
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In: ELECTROPHORESIS, Vol. 23, No. 4, 20.02.2002, p. 640-646.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Biochemical dissection of the mitochondrial proteome from Arabidopsis thaliana by three-dimensional gel electrophoresis
AU - Werhahn, Wolf
AU - Braun, Hans Peter
PY - 2002/2/20
Y1 - 2002/2/20
N2 - We report a subdivision of the mitochondrial proteome into defined sets of proteins, which is based on the combination of three different gel electrophoresis procedures. First, Blue-native polyacrylamide gel electrophoresis is employed to separate mitochondrial protein complexes. The protein complexes are electroeluted and completely detached from Coomasssie blue. Subsequently the subunits of the protein complexes are separated by isoelectric focusing and finally by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The resolution capacity of the procedure is demonstrated for the ATP synthase complex, the cytochrome c reductase complex and the preprotein translocase of the outer mitochondrial membrane (the TOM complex). The method allows the separation of isoforms of subunits forming part of protein complexes, whose occurrence seems to be rather a rule than an exception in higher eukaryotes. Furthermore, extremely hydrophobic proteins are detectable on the gels.
AB - We report a subdivision of the mitochondrial proteome into defined sets of proteins, which is based on the combination of three different gel electrophoresis procedures. First, Blue-native polyacrylamide gel electrophoresis is employed to separate mitochondrial protein complexes. The protein complexes are electroeluted and completely detached from Coomasssie blue. Subsequently the subunits of the protein complexes are separated by isoelectric focusing and finally by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The resolution capacity of the procedure is demonstrated for the ATP synthase complex, the cytochrome c reductase complex and the preprotein translocase of the outer mitochondrial membrane (the TOM complex). The method allows the separation of isoforms of subunits forming part of protein complexes, whose occurrence seems to be rather a rule than an exception in higher eukaryotes. Furthermore, extremely hydrophobic proteins are detectable on the gels.
KW - Arabidopsis thaliana
KW - Plant mitochondria
KW - Three-dimensional gel electrophoresis
UR - http://www.scopus.com/inward/record.url?scp=0036119404&partnerID=8YFLogxK
U2 - 10.1002/1522-2683(200202)23:4<640::AID-ELPS640>3.0.CO;2-F
DO - 10.1002/1522-2683(200202)23:4<640::AID-ELPS640>3.0.CO;2-F
M3 - Article
C2 - 11870776
AN - SCOPUS:0036119404
VL - 23
SP - 640
EP - 646
JO - ELECTROPHORESIS
JF - ELECTROPHORESIS
SN - 0173-0835
IS - 4
ER -