Elucidation of CPX‐1 involvement in RANKL‐induced osteoclastogenesis by a proteomics approach

FEBS Letters

Kim, Hyunsoo¹  Chang, Eun-Ju¹  Lee, Zang Hee³  Kwak, Han Bok³  Kim, Hong-Hee¹  Park, Joo-Cheol² 

[1] Department of Cell and Developmental Biology, Dental Research Institute, and Brain Korea 21 Program, College of Dentistry, Seoul National University, Seoul 110-749, South Korea;College of Dentistry, Chosun University, Gwangju 501-759, South Korea;National Research Laboratory for Bone Metabolism, Chosun University, Gwangju 501-759, South Korea

关键词: Osteoclast;    RANKL;    CPX-1;    Proteomics;    RANKL;    receptor activator of NF-κB ligand;    TRAP;    tartrate-resistant acid phosphatase;    2-DE;    two-dimensional gel electrophoresis;    MALDI-TOF;    matrix-assisted laser desorption/ionization-time of flight;   

DOI  :  10.1016/S0014-5793(04)00338-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To identify proteins potentially involved in osteoclast differentiation, we conducted a proteomics-based analysis using the osteoclastogenesis model cell line RAW264.7. Total proteins from undifferentiated cells, committed pre-osteoclasts, and differentiated osteoclasts were resolved by two-dimensional gel electrophoresis. Protein spots showing differential expression levels were processed for peptide mass fingerprinting. Among them, we identified the metallocarboxypeptidase CPX-1, which was prominently increased in pre-osteoclasts and then decreased in mature osteoclasts. Results of reverse transcription polymerase chain reaction, Western blot, and confocal microscopy were in agreement with the proteomics data. Notably, the forced overexpression of CPX-1 led to the inhibition of osteoclast formation, but not pre-osteoclast generation. Therefore, the transient up-regulation pattern of CPX-1 expression may be important for the successful progression from pre-osteoclasts to mature osteoclasts.

【 授权许可】

Unknown