Centre for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen , Øster Voldgade 5-7, 1350, Copenhagen, Denmark.
J Proteome Res. 2012 Feb 3;11(2):917-26. doi: 10.1021/pr200721u. Epub 2011 Dec 14.
We used high-sensitivity, high-resolution tandem mass spectrometry to shotgun sequence ancient protein remains extracted from a 43 000 year old woolly mammoth ( Mammuthus primigenius ) bone preserved in the Siberian permafrost. For the first time, 126 unique protein accessions, mostly low-abundance extracellular matrix and plasma proteins, were confidently identified by solid molecular evidence. Among the best characterized was the carrier protein serum albumin, presenting two single amino acid substitutions compared to extant African ( Loxodonta africana ) and Indian ( Elephas maximus ) elephants. Strong evidence was observed of amino acid modifications due to post-mortem hydrolytic and oxidative damage. A consistent subset of this permafrost bone proteome was also identified in more recent Columbian mammoth ( Mammuthus columbi ) samples from temperate latitudes, extending the potential of the approach described beyond subpolar environments. Mass spectrometry-based ancient protein sequencing offers new perspectives for future molecular phylogenetic inference and physiological studies on samples not amenable to ancient DNA investigation. This approach therefore represents a further step into the ongoing integration of different high-throughput technologies for identification of ancient biomolecules, unleashing the field of paleoproteomics.
我们使用高灵敏度、高分辨率串联质谱技术对保存在西伯利亚永冻层中的一只 43000 年前的长毛猛犸象（Mammuthus primigenius）骨骼中提取的古代蛋白质进行了鸟枪法测序。这是首次通过确凿的分子证据鉴定出 126 种独特的蛋白质，这些蛋白质主要是低丰度的细胞外基质和血浆蛋白。其中特征最为明显的是载脂蛋白血清白蛋白，与现存的非洲象（Loxodonta africana）和印度象（Elephas maximus）相比，它有两个单一氨基酸的替换。大量证据表明，由于死后水解和氧化损伤，氨基酸发生了修饰。在来自温带地区的更近的哥伦比亚猛犸象（Mammuthus columbi）样本中，也发现了这种永冻土骨骼蛋白质组的一个一致子集，这将该方法的应用范围扩展到了亚极地区域之外。基于质谱的古代蛋白质测序为未来对不适于进行古 DNA 研究的样本进行分子系统发育推断和生理学研究提供了新的视角。因此，这种方法代表了不同高通量技术在鉴定古代生物分子方面的不断整合的又一步，释放了古蛋白质组学领域的潜力。
