Engineering and refolding of a novel trimeric fusion protein TRAIL-collagen XVIII NC1
Engineering and refolding of a novel trimeric fusion protein TRAIL-collagen XVIII NC1
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新型三聚体融合蛋白 TRAIL-胶原 XVIII NC1 的工程设计和重折叠
DOI：
10.1007/s00253-012-4604-0
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发表时间：
2013-08
影响因子：
5
通讯作者：
Chen, Shu Qing
中科院分区：
文献类型：
--
作者：
Pan, Li Qiang;Xie, Zhang Ming;Tang, Xiao Jing;Wu, Min;Wang, Fu Rong;Naranm;ura, Hua;Chen, Shu Qing
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 inEscherichia colias 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.
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影响因子：
2.9
作者：
Vagenende, Vincent;Yap, Miranda G. S.;Trout, Bernhardt L.
通讯作者：
Trout, Bernhardt L.
影响因子：
5.6
作者：
Boudko SP;Sasaki T;Engel J;Lerch TF;Nix J;Chapman MS;Bächinger HP
通讯作者：
Bächinger HP
影响因子：
3.8
作者：
B. Batas;J. Chaudhuri
通讯作者：
B. Batas;J. Chaudhuri
影响因子：
46.9
作者：
Günter Stempfer;B. Höll-Neugebauer;R. Rudolph
通讯作者：
Günter Stempfer;B. Höll-Neugebauer;R. Rudolph
影响因子：
4.1
作者：
J. Ahn;Young Phil Lee;J. Rhee
通讯作者：
J. Ahn;Young Phil Lee;J. Rhee