In-depth analysis of the membrane and cytosolic proteome of red blood cells
In-depth analysis of the membrane and cytosolic proteome of red blood cells
复制标题
DOI：
10.1182/blood-2005-11-007799
复制
发表时间：
2006-08-01
期刊：
影响因子：
20.3
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Pasini, Erica M.;Kirkegaard, Morten;Mann, Matthias
除了将氧气和二氧化碳运输到组织和从组织运输之外，脊椎动物的红细胞（RBC）还具有一系列其他功能。在这些功能中的任何一个中损害RBC性能的疾病都需要深入研究。此外，人类红细胞是疟原虫的重要宿主细胞。从经典的生物化学方法中已经了解到许多关于RBC组成和膜组织的信息。在这里，我们使用质谱（MS）为基础的蛋白质组学来表征正常红细胞蛋白质谱。本研究的目的是获得最完整和信息丰富的人类红细胞蛋白质组可能结合高精度，高灵敏度的蛋白质鉴定技术（四极杆飞行时间和傅立叶变换MS）与选定的生化程序的样品制备。共鉴定、验证了340种膜蛋白和252种可溶性蛋白，并根据亚细胞定位、蛋白质家族和功能进行了分类。剪接异构体的蛋白质被确定，和多肽，迁移与令人不安的高或低的表观分子量可以被分组到泛素化，部分降解，或酯连接的复合物。我们的数据揭示了RBC蛋白质组的意外复杂性，提供了丰富的数据，其组成，揭示了RBC生物学中的几个开放问题，并形成了全面了解RBC功能的出发点。
In addition to transporting oxygen and carbon dioxide to and from the tissues, a rang a of other functions are attributed to red blood cells (RBCs) of vertebrates. Diseases compromising RBC performani.e in any of these functions warrant in-depth study. Furthermore, the human RBC is a vital host cell for the malaria parasite. Much has been learned from classical biochemical approaches about RBC composition and membrane organization. Here, we use mass spectrometry (MS)-based proteomics to characterize the normal RBC protein profile. The aim of this study was to obtain the most complete and informative human RBC proteome possible by combining highaccuracy, high-sensitivity protein identification technology (quadrupole time of flight and Fourier transform MS) with selected biochemical procedures for sample preparation. A total of 340 membrane proteins and 252 soluble proteins were identified, validated, and categorized in terms of subcellular localization, protein family, and function. Splice isoforms of proteins were identified, and polypeptides that migrated with anomalously high or low apparent molecular weights could be grouped into either ubiquitinylated, partially degraded, or ester-linked complexes. Our data reveal unexpected complexity of the RBC proteome, provide a wealth of data on its composition, shed light on several open issues in RBC biology, and form a departure point for comprehensive understanding of RBC functions.