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Biochemical dissection of the mitochondrial proteome from Arabidopsis thaliana by three‐dimensional gel electrophoresis
Author(s) -
Werhahn Wolf,
Braun HansPeter
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200202)23:4<640::aid-elps640>3.0.co;2-f
Subject(s) - isoelectric focusing , gel electrophoresis , proteome , polyacrylamide gel electrophoresis , two dimensional gel electrophoresis , biochemistry , gel electrophoresis of proteins , biology , membrane protein , chemistry , electrophoresis , proteomics , enzyme , gene , membrane
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.