title: In-depth analysis of cysteine oxidation by the RBC proteome: advantage of peroxiredoxin II knockout mice.
abstract: Peroxiredoxin II (Prdx II, a typical 2-Cys Prdx) has been originally isolated from erythrocytes, and its structure and peroxidase activity have been adequately studied. Mice lacking Prdx II proteins had heinz bodies in their peripheral blood, and morphologically abnormal cells were detected in the dense red blood cell (RBC) fractions, which contained markedly higher levels of reactive oxygen species (ROS). In this study, a labeling experiment with the thiol-modifying reagent biotinylated iodoacetamide (BIAM) in Prdx II-/- mice revealed that a variety of RBC proteins were highly oxidized. To identify oxidation-sensitive proteins in Prdx II-/- mice, we performed RBC comparative proteome analysis in membrane and cytosolic fractions by nano-UPLC-MSE shotgun proteomics. We found oxidation-sensitive 54 proteins from 61 peptides containing cysteine oxidation, and analyzed comparative expression pattern in healthy RBCs of Prdx II+/+ mice, healthy RBCs of Prdx II-/- mice, and abnormal RBCs of Prdx II-/- mice. These proteins belonged to cellular functions related with RBC lifespan maintain, such as cytoskeleton, stress-induced proteins, metabolic enzymes, signal transduction, and transporters. Furthermore, protein networks among identified oxidation-sensitive proteins were analyzed to associate with various diseases. Consequently, we expected that RBC proteome might provide clues to understand redox-imbalanced diseases.
authors: Department of Oral Biochemistry, Dental Science Research Institute, The 2nd Stage of Brain Korea 21 for Dental School, Chonnam National University, Gwangju, Republic of Korea.
authors: Hee-Young
authors: HY
authors: Yang
authors: Joseph
authors: J
authors: Kwon
authors: Hoon-In
authors: HI
authors: Choi
authors: Seong Hwa
authors: SH
authors: Park
authors: Ung
authors: U
authors: Yang
authors: Hyang-Rim
authors: HR
authors: Park
authors: Lina
authors: L
authors: Ren
authors: Kyoung-Jin
authors: KJ
authors: Chung
authors: Youn U
authors: YU
authors: Kim
authors: Byung-Ju
authors: BJ
authors: Park
authors: Sang-Hun
authors: SH
authors: Jeong
authors: Tae-Hoon
authors: TH
authors: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006706:Homeostasis; D007460:Iodoacetamide / Q000737:chemistry; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry; D054464:Peroxiredoxins / Q000235:genetics* / Q000378:metabolism; D060066:Protein Interaction Maps; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003598:Cytoskeletal Proteins; D010446:Peptide Fragments; D020543:Proteome; D054464:Peroxiredoxins; D003545:Cysteine; D007460:Iodoacetamide