Schägger H, Bentlage H, Ruitenbeek W, Pfeiffer K, Rotter S, Rother C, Böttcher-Purkl A, Lodemann E
Zentrum der Biologischen Chemie, Universitätsklinikum Frankfurt, Germany.
Electrophoresis. 1996 Apr;17(4):709-14. doi: 10.1002/elps.1150170415.
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.
一种将蓝色天然聚丙烯酰胺凝胶电泳（BN-PAGE）与十二烷基硫酸钠-三羟甲基氨基甲烷（SDS）-PAGE相结合的二维电泳技术，先前已用于从活检或尸检组织开始的人类疾病中氧化磷酸化（OXPHOS）缺陷的定位（沙格，H.，《电泳》，1995年，16卷，763 - 770页）。在本研究中，该技术被扩展用于从血小板和组织培养细胞中分离氧化磷酸化酶。在二维凝胶中检测氧化磷酸化复合物的蛋白质亚基需要银染。然而，使用培养细胞对线粒体脑肌病患者有重大影响，因为这将减少侵入性肌肉活检的次数。从几毫升血液中轻松分离血小板膜糖蛋白复合物，使得在诸如血小板无力症等出血性疾病中分析该复合物及其蛋白质亚基成为可能。
