Department of Molecular Genetics, Room 4280 Medical Sciences Building, 1 King's College Circle, Toronto, ON M5S1A8, Canada.
Fungal Genet Biol. 2011 May;48(5):526-36. doi: 10.1016/j.fgb.2010.11.011. Epub 2010 Dec 7.
We have used gel electrophoretic techniques including isoelectric focusing, blue native, acid-urea, 16-benzyldimethyl-n-hexadecylammonium chloride or SDS first dimensions and SDS Laemmli or tricine second dimensions to separate the proteins from highly-purified Neurospora mitochondria and sub-mitochondrial fractions (membrane, soluble, protein complexes and ribonucleoproteins). The products of 260 genes, many of them in multiple processed or modified forms, were identified by MALDI-TOF mass spectrometry. This work confirms the existence, expression, and mitochondrial localization of the products of 55 Neurospora genes previously annotated only as predicted or hypothetical, and of 101 genes not identified in previous mass spectrometry studies. Combined with previous mass spectrometry studies, and re-evaluation of genome annotations, we have compiled a curated list of 358 proteins identified in proteomic studies that are likely to be bona fide mitochondrial proteins plus 80 other identified proteins that may be mitochondrial. Literature data mining and computational predictions suggest that Neurospora mitochondria also contain another 299 proteins not yet identified in proteomics projects. Taken together, these data suggest that the products of at least 738 genes comprise the Neurospora mitochondrial proteome.
我们已经使用凝胶电泳技术，包括等电聚焦、蓝色-native、酸-尿素、16-苄基二甲基-n-十六烷基氯化铵或 SDS 第一维以及 SDS Laemmli 或 tricine 第二维，从高度纯化的 Neurospora 线粒体和亚线粒体部分（膜、可溶性、蛋白质复合物和核糖核蛋白）中分离蛋白质。通过 MALDI-TOF 质谱鉴定了 260 个基因的产物，其中许多是经过多个处理或修饰形式的。这项工作证实了以前仅注释为预测或假设的 55 个 Neurospora 基因产物的存在、表达和线粒体定位，以及以前的质谱研究中未鉴定出的 101 个基因产物的存在、表达和线粒体定位。结合以前的质谱研究和对基因组注释的重新评估，我们编译了一份经过精心整理的 358 种蛋白质列表，这些蛋白质是在蛋白质组学研究中鉴定出来的，可能是真正的线粒体蛋白质，加上另外 80 种已鉴定的可能是线粒体的蛋白质。文献数据挖掘和计算预测表明，Neurospora 线粒体还含有另外 299 种尚未在蛋白质组学项目中鉴定出的蛋白质。总之，这些数据表明，Neurospora 线粒体蛋白质组至少由 738 个基因的产物组成。
