NDLI: Engineering and refolding of a novel trimeric fusion protein TRAIL-collagen XVIII NC1

Content Provider	Springer Nature Link
Author	Pan, Li Qiang Xie, Zhang Ming Tang, Xiao Jing Wu, Min Wang, Fu Rong Naranmandura, Hua Chen, Shu Qing
Copyright Year	2012
Abstract	Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered to be a promising anticancer agent because its active form TRAIL trimer is able to induce apoptosis in different tumor cell lines while sparing normal cells. However, TRAIL trimer possesses a short half-life and low stability, which turns out to be a major obstacle for the development of clinical trials. In our present study, we constructed a recombined TRAIL trimer by genetic fusion of non-collagenous domain (NC1) of human collagen XVIII or its trimerization domain (TD) to C-terminus of TRAIL via a flexible linker, and then refolded the fusion proteins using a two-step refolding approach, namely a combination of dilution and gel filtration chromatography. As a result, both recombinant proteins, TRAIL-NC1 and TRAIL-TD, were expressed in Escherichia coli as inclusion bodies, and they exhibited difficultly to refold efficiently by conventional methods. Thereby, we applied a modified two-step refolding approach to refold fusion proteins. More than 55 % of TRAIL-NC1 and 90 % of TRAIL-TD protein activity was recovered during the two-step refolding approach, and their stability was also increased significantly. Also, size exclusion chromatography showed refolded TRAIL-NC1 was a trimer while TRAIL-TD, hexamer. However, both of them exerted good apoptosis activity on NCI-H460 cells.
Starting Page	7253
Ending Page	7264
Page Count	12
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	97
Issue Number	16
e-ISSN	14320614
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2012-12-04
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	TRAIL Non-collagenous domain Protein refolding Apoptosis Two-step refolding approach Microbiology Microbial Genetics and Genomics Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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