Recombinant expression and purification of a MAP30-cell penetrating peptide fusion protein with higher anti-tumor bioactivity

Protein Expression and Purification
Qiang LvFu-Jun Wang

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 improv...Continue Reading

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Citations

Oct 16, 2016·Journal of Peptide Science : an Official Publication of the European Peptide Society·Zhao LuoFu-Jun Wang
Nov 4, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yun SunYao Meng
Jun 19, 2018·Anti-cancer Drugs·Zhen-Tao SongFu-Jun Wang
Jul 17, 2019·Journal of Peptide Science : an Official Publication of the European Peptide Society·Yuan-Yuan XuJian Zhao
Oct 7, 2017·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yuping WeiXia Xu
Jan 28, 2021·Toxins·Lucía CitoresJosé M Ferreras
Jul 13, 2020·International Journal of Biological Macromolecules·Xiaojiao FanTengchuan Jin
May 3, 2021·Protein Expression and Purification·Wei-Wei ChenZhen-Hong Zhu

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