EXPRESSION, PURIFICATION, AND CHARACTERIZATION OF RGD-mda-7, A HIS-TAGGED mda-7/IL-24 MUTANT PROTEIN

• Published in: Journal of immunoassay & immunochemistry 2012-10, Vol.33 (4), p.352-368
• Main Authors: Liu, Jun-Jie, Zhang, Bao-Fu, Yin, Xiao-Xing, Pei, Dong-Sheng, Yang, Zhi-Xia, Di, Jie-Hui, Chen, Fei-Fei, Li, Hui-Zhong, Xu, Wei, Wu, Yong-Ping, Zheng, Jun-Nian
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents - isolation & purification; Antineoplastic Agents - pharmacology; antitumor; Apoptosis - drug effects; Cell Adhesion - drug effects; Cell Line, Tumor; Cell Survival - drug effects; Cloning, Molecular; Drug Screening Assays, Antitumor; Humans; Immunization; Integrin alphaVbeta3 - immunology; integrin αVβ; Interleukins - genetics; Interleukins - immunology; Interleukins - isolation & purification; MCF-7 Cells; mda-7/il-24; mutant; Mutation; Oligopeptides - genetics; Oligopeptides - isolation & purification; Oligopeptides - pharmacology; Receptors, Vitronectin - immunology; RGD; Structure-Activity Relationship; αVβ
• ISSN: 1532-1819; 1532-4230
• DOI: 10.1080/15321819.2012.659782

RGD peptide (Arg-Gly-Asp tripeptide) binds to integrin αVβ 3 and αVβ 5 , which is selectively expressed in tumor neovasculature and on the surface of some tumor cells. Some studies showed that coupling the RGD peptides to anticancer drugs yielded compounds with increased efficiency against tumors and lowered toxicity to normal tissues. The melanoma differentiation-associated gene-7/interleukin-24 gene (mda-7/IL-24) is a novel tumor-suppressor/cytokine gene that exhibits potent tumor-suppressive activity without damaging normal cells. To enhance the antitumor effect, we inserted a glycine residue into the wild type (mda-7/IL-24) between 164 Arg and 165 Asp to form a RGD peptide, named RGD-mda-7, then expressed RGD-mda-7 in Escherichia coli. Herein, we describe the expression and purification of RGD-mda-7. We detected the characterizations of immunostimulatory activity, tumor targeting, potent cytopathic effect, and apoptosis inducing exploited by RGD-mda-7 in tumor cells, and also compared these characterizations with wtmda-7/IL-24. The data showed that RGD-mda-7 had more potent tumor targeting and apoptosis-inducing effects than wtmda-7/IL-24.