Electrophoretic separation of multiprotein complexes from blood platelets and cell lines: technique for the analysis of diseases with defects in oxidative phosphorylation.
A two-dimensional electrophoretic technique combining blue native polyacrylamide gel electrophoresis (BN-PAGE) with Tricine sodium dodecyl sulfate (SDS)-PAGE was previously used for the localization of oxidative phosphorylation (OXPHOS) defects in human diseases starting from biopsy or autopsy tissues (Schägger, H., Electrophoresis 1995, 16, 763-770). In the present work the technique was extended for the resolution of OXPHOS enzymes from platelets and tissue-cultured cells. Silver staining is required to detect the protein subunits of OXPHOS complexes in two-dimensional gels. However, the use of cultured cells has major implications for patients with mitochondrial encephalomyopathies since it will reduce the number of invasive muscle biopsies. The ease of isolating the platelet membrane glycoprotein complex from a few milliliters of blood makes it possible to analyze this complex and its protein subunits in bleeding disorders like Glanzmann's thrombasthenia.
Schägger
H
H
Zentrum der Biologischen Chemie, Universitätsklinikum Frankfurt, Germany.
Bentlage
H
H
Ruitenbeek
W
W
Pfeiffer
K
K
Rotter
S
S
Rother
C
C
Böttcher-Purkl
A
A
Lodemann
E
E
D016475:3T3 Cells; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002460:Cell Line; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D010085:Oxidative Phosphorylation; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
D012967:Sodium Dodecyl Sulfate