Generic placeholder image

Protein & Peptide Letters

Editor-in-Chief

ISSN (Print): 0929-8665
ISSN (Online): 1875-5305

Fused hydrophobic elastin-like-peptides (ELP) enhance biological activity of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)

Author(s): Kaizong Huang, Ningjun Duan, Wenyan Zou, Chunmei Zhang, Yueyang Lai, Pingping Shen and Zichun Hua

Volume 22, Issue 11, 2015

Page: [1000 - 1006] Pages: 7

DOI: 10.2174/0929866522666150824162015

Price: $65

Abstract

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent with tumor-selective apoptotic activity. Formation of aggregates as trimer is the prerequisite for TRAIL’s function as an apoptosis inducer. However the polymerization property of TRAIL has also brought difficulties for its production. RGD-TRAIL is an integrin-targeting TRAIL mutant with enhanced apoptosisinducing activity towards tumor cells both in vitro and in vivo. When expressed in E. coli, TRAIL or its mutant RGDTRAIL usually formed inclusion bodies. Their extreme aggregation propensity for aggregation destabilizes the protein, leading to poor recovery and therefore low yield from the purification process. The low purification efficiency of TRAIL retards its industrial application and large-scale production. To avoid the above problems during RGD-TRAIL production, we employed elastin-like polypeptides (ELPs) for the fusion-expression of recombinant RGD-TRAIL. Recombinant RGD-TRAIL-ELP was expressed in a soluble form and efficiently purified from the clarified cell extracts by three rounds of inverse transition cycling (ITC). SDS–PAGE and Western blotting analyses of purified RGD-TRAIL-ELP showed that RGD-TRAIL-ELP was successfully purified and the yield was up to 10 mg/L of bacterial culture. Apoptosis assay was performed in human colorectal carcinoma cells (COLO-205) and human breast cancer cell line (MDA-MB-231) to assess the potency of the fusion protein. Fusion with hydrophobic ELP effectively enhanced RGD-TRAIL’s biological activity. The higher activity and appropriate particle size of RGD-TRAIL-ELP could be used for RGD-TRAIL delivery in tumor therapy.

Keywords: Biological activity, elastin-like polypeptide, homomultimer, inverse transition cycling, transition temperature, tumor necrosis factor-related apoptosis-inducing ligand.

Next »

Rights & Permissions Print Cite
© 2024 Bentham Science Publishers | Privacy Policy