title: In-depth analysis of the membrane and cytosolic proteome of red blood cells.
abstract: In addition to transporting oxygen and carbon dioxide to and from the tissues, a range of other functions are attributed to red blood cells (RBCs) of vertebrates. Diseases compromising RBC performance 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 high-accuracy, 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.
authors.affiliation: Department of Proteomics and Signal Transduction, Am Klopferspitz 18, D-82152 Martinsried, Germany.
authors.forename: Erica M
authors.initials: EM
authors.lastname: Pasini
authors.forename: Morten
authors.initials: M
authors.lastname: Kirkegaard
authors.forename: Peter
authors.initials: P
authors.lastname: Mortensen
authors.forename: Hans U
authors.initials: HU
authors.lastname: Lutz
authors.forename: Alan W
authors.initials: AW
authors.lastname: Thomas
authors.forename: Matthias
authors.initials: M
authors.lastname: Mann
mesh_terms: D003600:Cytosol / Q000737:chemistry*; D004910:Erythrocyte Membrane / Q000737:chemistry*; D004912:Erythrocytes / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008565:Membrane Proteins / Q000032:analysis* / Q000502:physiology; D020033:Protein Isoforms / Q000032:analysis; D011506:Proteins / Q000032:analysis* / Q000502:physiology; D020543:Proteome / Q000032:analysis* / Q000502:physiology; D040901:Proteomics / Q000379:methods; D012995:Solubility
chemical_list: D008565:Membrane Proteins; D020033:Protein Isoforms; D011506:Proteins; D020543:Proteome