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Updated: Oct 12, 2026
Engineering Cell-permeable Protein
Published on: December 28, 2009
Recombinant expression and purification of a MAP30-cell penetrating peptide fusion protein with higher anti-tumor
Qiang Lv1, Xu-Zhong Yang2, Long-Yun Fu3
1State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Abstract:
MAP30 (Momordica Antiviral Protein 30 Kd), a single-stranded type-I ribosome inactivating protein, possesses versatile biological activities including anti-tumor abilities. However, the low efficiency penetrating into tumor cells hampers the tumoricidal effect of MAP30. This paper describes MAP30 fused with a human-derived cell penetrating peptide HBD which overcome the low uptake efficiency by tumor cells and exhibits higher anti-tumor bioactivity. MAP30 gene was cloned from the genomic DNA of Momordica charantia and the recombinant plasmid pET28b-MAP30-HBD was established and transferred into Escherichia coli BL21 (DE3). The recombinant MAP30-HBD protein (rMAP30-HBD) was expressed in a soluble form after being induced by 0.5mM IPTG for 14h at 15°C. The recombinant protein was purified to greater than 95% purity with Ni-NTA affinity chromatography. The rMAP30-HBD protein not only has topological inactivation and protein translation inhibition activity but also showed significant improvements in cytotoxic activity compared to that of the rMAP30 protein without HBD in the tested tumor cell lines, and induced higher apoptosis rates in HeLa cells analyzed by Annexin V-FITC with FACS. This paper demonstrated a new method for improving MAP30 protein anti-tumor activity and might have potential applications in cancer therapy area.
Insights
Momordica Antiviral Protein 30 Kd (MAP30) fused with a cell-penetrating peptide (HBD) enhances anti-tumor efficacy. This novel fusion protein (rMAP30-HBD) improves tumor cell uptake and boosts anti-cancer activity, showing promise for cancer therapy.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Momordica Antiviral Protein 30 Kd (MAP30) is a type-I ribosome inactivating protein with demonstrated anti-tumor properties.
- Limited tumor cell penetration restricts the therapeutic potential of MAP30.
- Cell-penetrating peptides offer a strategy to enhance protein delivery into target cells.
Purpose of the Study:
- To develop a fusion protein combining MAP30 with a human-derived cell-penetrating peptide (HBD).
- To evaluate the enhanced anti-tumor activity and cellular uptake of the recombinant MAP30-HBD (rMAP30-HBD) protein.
- To assess the potential of rMAP30-HBD as a therapeutic agent for cancer treatment.
Main Methods:
- Cloning of the MAP30 gene from Momordica charantia and construction of the pET28b-MAP30-HBD plasmid.
- Expression and purification of the recombinant rMAP30-HBD protein in E. coli using Ni-NTA affinity chromatography.
- Assessment of cytotoxic activity and apoptosis induction in tumor cell lines, including HeLa cells, using Annexin V-FITC and FACS analysis.
Main Results:
- The recombinant rMAP30-HBD protein was successfully expressed and purified to >95% purity.
- rMAP30-HBD exhibited enhanced cytotoxic activity against tested tumor cell lines compared to MAP30 alone.
- rMAP30-HBD induced significantly higher apoptosis rates in HeLa cells than the non-fused MAP30 protein.
Conclusions:
- Fusion of MAP30 with the HBD peptide effectively overcomes the low tumor cell uptake limitation.
- The rMAP30-HBD fusion protein demonstrates improved anti-tumor bioactivity and apoptosis-inducing potential.
- This strategy presents a promising approach for enhancing MAP30-based cancer therapy.
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