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. Prdx Ⅱ has been reported to protect a wide range of cellular environments as antioxidant enzyme, and its dysfunctions may be implicated in a variety of disease states associated with oxidative stress, including cancer and aging-associated pathologies. But, the precise mechanism is still obscure in various aspects of aging containing ovarian aging. Identification and relative quantification of the increased proteins affected by Prdx Ⅱ deficiency may help identify novel signaling mechanisms that are important for oxidative stress-related diseases. To identify the increased proteins in Prdx Ⅱ—/— mice, we performed RBC comparative proteome analysis in membrane fraction and cytosolic fractions by nano-UPLC-MSE shotgun proteomics. We found the increased 86 proteins in membrane (32 proteins) and cytosolic (54 proteins) fractions, 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 cellular morphology and assembly, cell-cell interaction, metabolism, and stress-induced signaling. Moreover, protein networks among the increased proteins were analyzed to associate with various diseases. Taken together, RBC proteome may provide clues to understand the clue about redox-imbalanced diseases.

ABSTRACT   INTRODUCTION   MATERIALS AND METHODS    Animals    RBC Fractionation    The RBC Proteome    Nano-UPLC-MSE Tandem Mass Spectrometry    Protein Identification and Quantitative Analysis    Bioinformatics Analysis    Western Blotting   RESULTS    Identification of Proteins Differentially Regulated in Prdx Ⅱ Deficient RBCs    Protein Network Comprised of Identified Proteins RegulatedBy Prdx Ⅱ Deficiency    Verification of the Expressional Changes in Prdx Ⅱ—/—RBCs   DISCUSSION   REFERENCES

• Hee-Young Yang(Department of Oral Biochemistry, Dental Science Research Institute and the BK21 Project, Medical Research Center for Biomineralization, Disorders, School of Dentistry, Chonnam National University, Gwangju 500-757, Korea.)
• Tae-Hoon Lee(Department of Oral Biochemistry, Dental Science Research Institute and the BK21 Project, Medical Research Center for Biomineralization, Disorders, School of Dentistry, Chonnam National University, Gwangju 500-757, Korea.) Corresponding author