Article ID Journal Published Year Pages File Type
2020269 Protein Expression and Purification 2016 8 Pages PDF
Abstract

•This is the first report of fusion protein TRAIL-CM4 expression in Escherichia coli.•TRAIL-CM4 exhibits stronger anti-proliferative activity against tumor cells than solo TRAIL.•TRAIL-CM4 also induces more tumor cells apoptosis than solo TRAIL.•TRAIL-CM4 exerts strong antibacterial activity like CM4.

A TRAIL-CM4 fusion protein in soluble form with tumor selective apoptosis and antibacterial functions was expressed in the Escherichia coli expression system and isolated through dialysis refolding and histidine-tag Nickel-affinity purification. Fresh Jurkat cells were treated with the TRAIL-CM4 fusion protein. Trypan blue staining and MTT analyses showed that, similar to a TRAIL positive control, Jurkat cell proliferation was significantly inhibited. Flow cytometry analyses using Annexin V-fluorescein revealed that Jurkat cells treated with the TRAIL-CM4 fusion protein exhibited increased apoptosis. Laser confocal microscopy showed that APB-CM4 and the fusion protein TRAIL-CM4 can bind to Jurkat cell membranes and initiate their destruction. ABP-CM4 enhances the antitumor activity of TRAIL by targeting and damaging the tumor cell membrane. In antibacterial experiments, agar well diffusion and bacterial growth inhibition curve assays revealed concentration-dependent TRAIL-CM4 antibacterial activity against Escherichia coli K12D31. The expressed TRAIL-CM4 fusion protein exhibited enhanced antitumor and antibacterial activities. Fusion protein expression allowed the two different proteins to function in combination.

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Life Sciences Biochemistry, Genetics and Molecular Biology Biochemistry
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