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SAC-TRAIL, a novel anticancer fusion protein: expression, purification, and functional characterization
Jian Zhang1, Wanyuan Dong2, Yuhong Ren2
1State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, People's Republic of China. zhangjian0020@ecust.edu.cn.
A novel fusion protein, SAC-TRAIL, enhances cancer cell apoptosis and suppresses tumor growth more effectively than native TRAIL. This improved therapeutic potential stems from its specific cancer cell targeting and optimized structural properties.
Area of Science:
- Biotechnology
- Molecular Biology
- Cancer Research
Background:
- Recombinant proteins are vital in cancer therapy, but tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) efficacy is limited by drug resistance.
- Prostate apoptosis response-4 (Par-4) selectively induces apoptosis in cancer cells via GRP78 receptor binding.
Purpose of the Study:
- To develop a novel recombinant fusion protein combining TRAIL and Par-4 (SAC) to overcome TRAIL resistance in cancer treatment.
- To optimize the fusion protein's structure and assess its enhanced antitumor activity and specificity.
Main Methods:
- A fusion protein, SAC-TRAIL, was constructed with a small ubiquitin-related modifier (SUMO) tag for solubility.
- Cytotoxicity, immunofluorescence, and circular dichroism assays were employed to evaluate the fusion protein's efficacy, specificity, and structural characteristics.
Main Results:
- The purified SAC-TRAIL fusion protein demonstrated superior antitumor activity compared to native TRAIL.
- The fusion protein with a flexible (G4S)3 linker exhibited optimal inhibition of cancer cell proliferation and specific binding to cancer cells.
- Circular dichroism revealed that the flexible linker promoted a structure with lower alpha-helix and higher random coiling, facilitating receptor binding.
Conclusions:
- The novel recombinant fusion protein SAC-(G4S)3-TRAIL represents a promising therapeutic agent for cancer treatment.
- The fusion protein exhibits enhanced tumor growth suppression and apoptosis induction, with improved targeting selectivity in cancer cells.
