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.
Red blood cell Proteome Membranar proteins Mass spectrometry Proteases Infection
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RM is supported by FCT investigator program 2012 (IF/01002/2012). ASC is supported by grant SFRH/BPD/85569/2012 funded by Fundação para a Ciência e Tecnologia.
Ringrose JH et al (2008) Highly efficient depletion strategy for the two most abundant erythrocyte soluble proteins improves proteome coverage dramatically. J Proteome Res 7:3060–3063. doi:10.1021/pr8001029CrossRefPubMedGoogle Scholar
Feliciano A et al (2016) Evening-morning peroxiredoxin-2 redox/oligomeric state changes in obstructive sleep apnea red blood cells: correlation with polysomnographic and metabolic parameters. Biochim Biophys Acta 1863(2):621–629. doi:10.1016/j.bbadis.2016.11.019CrossRefPubMedGoogle Scholar
Wisniewski JR, Zougman A, Mann M (2009) Combination of FASP and StageTip-based fractionation allows in-depth analysis of the hippocampal membrane proteome. J Proteome Res 8:5674–5678. doi:10.1021/pr900748nCrossRefPubMedGoogle Scholar
Cox J, Mann M (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26:1367–1372. doi:10.1038/nbt.1511CrossRefPubMedGoogle Scholar
Hegedus T et al (2015) Inconsistencies in the red blood cell membrane proteome analysis: generation of a database for research and diagnostic applications. Database (Oxford) 2015:bav056. doi:10.1093/database/bav056CrossRefGoogle Scholar