The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification
The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification
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DOI：
10.1074/jbc.c300064200
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发表时间：
2003-04-18
影响因子：
4.8
通讯作者：
Capaldi, RA
中科院分区：
文献类型：
--
作者：
Murray, J;Zhang, B;Capaldi, RA
NADH脱氢酶复合体的缺陷主要表现在线粒体疾病中，并与许多晚发型神经系统疾病的发展密切相关，如帕金森氏病。在这里，我们描述了一种从人体组织中分离这种多亚单位膜结合复合体的免疫捕获程序。利用少量的免疫分离蛋白、一维和二维凝胶电泳法、基质辅助激光解吸电离飞行时间(MALDI-TOF)多肽指纹图(PMF)和纳米流液相色谱质谱质谱(LCMS/MS)，我们可以分离和鉴定迄今在更广泛研究的牛心复合体1中检测到的42种多肽的人类同源物。这些多肽包括被认为参与细胞凋亡的GRIM-19蛋白，一种最先通过基因筛选a的神经元蛋白鉴定的多肽，以及一种被认为与分化相关的蛋白质。通过不同程序分离的人和牛复合体I的数据的一致性增加了这样一种确定性，即这些看似不同功能的新蛋白质是复合体1的一部分。
Defects of the NADH dehydrogenase complex are predominantly manifested in mitochondrial diseases and are significantly associated with the development of many late onset neurological disorders such as Parkinson's disease. Here we describe an immunocapture procedure for isolating this multisubunit membrane-bound complex from human tissue. Using small amounts of immunoisolated protein, one-dimensional and two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) peptide mass finger printing (PMF), and nanoflow liquid chromatography mass spectrometry/mass spectrometry (LCMS/MS), we can resolve and identify the human homologues of 42 polypeptides detected so far in the more extensively studied beef heart complex 1. These polypeptides include the GRIM-19 protein, which is claimed to be involved in apoptosis, a polypeptide first identified by gene screening a's a neuronal protein, as well as a protein thought to be in differentiation linked processes. The concordance of data from human and bovine complex I isolated by different procedures adds to the certainty that these novel proteins of seemingly diverse function are a part of complex 1.