文献类型：期刊文献

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

作者：Lv, Qiang[1];Yang, Xu-Zhong[1,2];Fu, Long-Yun[2];Lu, Yv-Ting[2];Lu, Yan-Hua[1];Zhao, Jian[1];Wang, Fu-Jun[2,3]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Zhejiang Reachall Pharmaceut Co Ltd, Dongyang 322100, Zhejiang, Peoples R China;[3]Shanghai Univ Chinese Tradit Med, Shanghai 201203, Peoples R China

年份：2015

卷号：111

起止页码：9

外文期刊名：PROTEIN EXPRESSION AND PURIFICATION

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000354148800002)】；

基金：This work was sponsored by the National Natural Science Foundation of China (Grant No. 81171449) and was partially supported by the Fundamental Research Funds for the Central Universities (WF1113010).

语种：英文

外文关键词：Recombinant Momordica Antiviral Protein 30 Kd (rMAP30); Ribosome inactivating protein; Cell Penetrating Peptide; Ni-NTA purification; Anti-tumor activity

摘要：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 HOD 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 0.5 mM IPTG for 14 h at 15 degrees 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. (C) 2015 Elsevier Inc. All rights reserved.