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

verfasst von

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.

Organisationseinheit(en)

Institut für Pflanzengenetik

Typ

Artikel

Journal

ELECTROPHORESIS

Band

23

Seiten

640-646

Anzahl der Seiten

7

ISSN

0173-0835

Publikationsdatum

20.02.2002

Publikationsstatus

Veröffentlicht

Peer-reviewed

Ja

ASJC Scopus Sachgebiete

Analytische Chemie, Biochemie, Klinische Biochemie

Elektronische Version(en)

https://doi.org/10.1002/1522-2683(200202)23:4<640::AID-ELPS640>3.0.CO;2-F (Zugang: Unbekannt)