Carvalho Ana Sofia, Rodriguez Manuel S, Matthiesen Rune
Computational and Experimental Biology Group, Nova Medical School/ Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Rua Câmara Pestana, 6-6A, Lisboa, 1150-082, Portugal.
Advanced Technology Institute in Life Sciences (ITAV), CNRS-USR3505, 31106, Toulouse, France.
Methods Mol Biol. 2017;1619:173-181. doi: 10.1007/978-1-4939-7057-5_13.
Red blood cells (RBCs) are known for their role in oxygen and carbon dioxide transport. The main function of RBCs is directly linked to many diseases that cause low oxygen levels in tissues such as congenital heart disease in adults, chronic obstructive pulmonary disease, sleep apnea, sickle cell disease, etc. Red blood cells are a direct target for a number of parasitic diseases such as malaria (Plasmodium) and similar parasites of the phylum Apicomplexa (Toxoplasma, Theileria, Eimeria, Babesia, and Cryptosporidium). RBC membrane components, in particular, are suitable targets for the discovery of drugs against parasite interaction. There is also evidence that RBCs release growth and survival factors, thereby linking RBCs with cancer. RBCs are abundant and travel throughout the body; consequently changes in RBC proteome potentially reflect other diseases as well. This chapter describes erythrocyte isolation from blood and its fractionation into RBC membrane and soluble cytosolic fractions. Alternative procedures for mass spectrometry analysis of RBC membrane proteome will be presented.
红细胞(RBCs)以其在氧气和二氧化碳运输中的作用而闻名。红细胞的主要功能与许多导致组织中氧含量低的疾病直接相关,如成人先天性心脏病、慢性阻塞性肺疾病、睡眠呼吸暂停、镰状细胞病等。红细胞是多种寄生虫病的直接靶点,如疟疾(疟原虫)以及顶复门的类似寄生虫(弓形虫、泰勒虫、艾美球虫、巴贝斯虫和隐孢子虫)。特别是红细胞膜成分,是发现抗寄生虫相互作用药物的合适靶点。也有证据表明红细胞会释放生长和存活因子,从而将红细胞与癌症联系起来。红细胞数量丰富且遍布全身;因此,红细胞蛋白质组的变化也可能反映其他疾病。本章描述了从血液中分离红细胞及其分离成红细胞膜和可溶性胞质部分的过程。还将介绍红细胞膜蛋白质组质谱分析的替代方法。