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Electrophoretic separation of multiprotein complexes from blood platelets and cell lines: Technique for the analysis of diseases with defects in oxidative phosphorylation
Author(s) -
Schägger Hermann,
Bentlage Herman,
Ruitenbeek Wim,
Pfeiffer Kathy,
Rotter Stefan,
Rother Christof,
BöttcherPurkl Andrea,
Lodemann Edgar
Publication year - 1996
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/elps.1150170415
Subject(s) - oxidative phosphorylation , gel electrophoresis , platelet , microbiology and biotechnology , biochemistry , polyacrylamide gel electrophoresis , chemistry , biology , enzyme , immunology
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.