Biochemical dissection of the mitochondrial proteome from Arabidopsis thaliana by three-dimensional gel electrophoresis

authored by: Wolf Werhahn, Hans Peter Braun
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.
Organisation(s): Institute of Plant Genetics
Type: Article
Journal: ELECTROPHORESIS
Volume: 23
Pages: 640-646
No. of pages: 7
ISSN: 0173-0835
Publication date: 20.02.2002
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Analytical Chemistry, Biochemistry, Clinical Biochemistry
Electronic version(s): https://doi.org/10.1002/1522-2683(200202)23:4<640::AID-ELPS640>3.0.CO;2-F (Access: Unknown)