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Construction and Expression of sTRAIL-Melittin CombiningEnhanced Anticancer Activity with Antibacterial Activity in escherichia coli
Appl Microbiol Biotechnol, 2012 Nov 13Tumor necrosis factor-related apoptosis-inducing ligand(TRAIL), as an anticancer protein with tumor-selective apoptotic activity, hasbeen examined for use in clinical application. Melittin, an antibacterialpeptide isolated from the bee Apis mellifera, has shown strong cytotoxicity toboth tumor and normal cells. To ameliorate the cytotoxicity of melittin oncells and enhance the activity of TRAIL on cancer cells, we constructed a novelfusion protein, sTRAIL-melittin, containing a small ubiquitin-related modifier(SUMO) tag and expressed this fusion protein in Escherichia coli. Data showedthat expression of the soluble fusion protein with the SUMO tag wasapproximately 85 % of total target protein which was much higher than thatwithout the SUMO tag (approximately 10 %); sTRAIL-melittin was easily purifiedusing Ni-NTA affinity chromatography and the tag was removed easily using SUMO-specificprotease. To assay anticancer activity and side effects, methyl thiazolyltetrazolium, hemolytic, and apoptosis assays were employed. Resultsdemonstrated that sTRAIL-melittin had cytotoxic and apoptotic activity in K562leukemia cells and HepG2 liver carcinoma cells, while it had only a minimaleffect on erythrocytes and normal HEK293 cells. This indicates that thecytotoxicity of sTRAIL-melittin in normal cells was low and the anticanceractivity of the fusion protein in tumor cells was significantly enhancedcompared with sTRAIL (P < 0.01). Furthermore, we found that sTRAIL-melittinalso showed antibacterial activity to Staphylococcus aureus due to the presenceof the melittin domain. Therefore, TRAIL fused with an antibacterial peptide maybe a promising novel TRAIL-based anticancer treatment strategy.
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