[Characterization of the interaction between low density lipoprotein receptor-related protein-associated protein 1 and the cancer and embryo expression protein 65]
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Published on: March 3, 2011
Geng-lin Jin1, Jian-zhi Zhang, Rong Su
1Peking University School of Oncology, Beijing Cancer Hospital, Beijing 100034, China.
Objective:
To understand the mechanism of cancer and embryo expression protein 65 (CEP65) in cancer development.
Methods:
CEP65 was used as a bait protein to isolate the partners of CEP65 from a human placenta cDNA library with yeast two hybrid system. The DNA fragments from positive clone were expressed prokaryotic and mammalian cells respectively, the GST pull-down experiment was performed to ascertain the binding activity.
Results:
After screening a human placenta cDNA library, the low density lipoprotein receptor-related protein-associated protein 1 (RAP) was isolated and shown to interact with CEP65. GST pull-down assay results indicated that His-CEP65 and GST-RAP expressed by prokaryotic system were immunoprecipitated, and so were Myc-RAP and GST-P65 expressed by mammalian cells.
Conclusion:
RAP can interact with CEP65 and RAP may be a candidate to mediate the function of CEP65.
Insights
Cancer and embryo expression protein 65 (CEP65) interacts with low-density lipoprotein receptor-associated protein 1 (RAP). This interaction suggests RAP may mediate CEP65 functions in cancer development, offering new insights into cancer mechanisms.
Area of Science:
- Molecular Biology
- Cancer Research
- Protein Interactions
Background:
- Cancer and embryo expression protein 65 (CEP65) is implicated in cancer development.
- Understanding CEP65's molecular interactions is crucial for elucidating its role in oncogenesis.
Purpose of the Study:
- To identify proteins interacting with CEP65.
- To investigate the functional relationship between CEP65 and its binding partners in the context of cancer.
Main Methods:
- Yeast two-hybrid system was employed to screen a human placenta cDNA library for CEP65 interacting partners.
- Prokaryotic and mammalian cell expression systems were used to validate protein interactions.
- GST pull-down assays were performed to confirm binding activity between CEP65 and identified partners.
Main Results:
- The study successfully isolated low-density lipoprotein receptor-associated protein 1 (RAP) as a binding partner of CEP65.
- GST pull-down assays confirmed the direct interaction between CEP65 and RAP in both prokaryotic and mammalian expression systems.
Conclusions:
- Low-density lipoprotein receptor-associated protein 1 (RAP) directly interacts with cancer and embryo expression protein 65 (CEP65).
- RAP is identified as a potential mediator of CEP65's function, suggesting a novel pathway in cancer development.
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