GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli

Abstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been...

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Auteurs principaux:	Zhanqing Wang, Min Zhang, Xin Lv, Jiying Fan, Jian Zhang, Jing Sun, Yaling Shen
Format:	article
Langue:	EN
Publié:	Nature Portfolio 2018
Sujets:	Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) Soluble TRAIL rhTRAIL GroE Chaperonin Coli Strain C600 Medicine R Science Q
Accès en ligne:	https://doaj.org/article/9c001b387a9d4afc83e57e681db6a0bc
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spelling	oai:doaj.org-article:9c001b387a9d4afc83e57e681db6a0bc2021-12-02T15:08:49ZGroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli10.1038/s41598-018-34090-72045-2322https://doaj.org/article/9c001b387a9d4afc83e57e681db6a0bc2018-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-34090-7https://doaj.org/toc/2045-2322Abstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been thoroughly studied. Here, a GroEL-GroES co-expressing strain and a deficient strain were constructed to study the in vivo recovery of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The interaction between GroEL/ES and TRAIL was simulated by molecular docking and identified by co-immunoprecipitation. The in vitro cytotoxicity of TRAIL IBs before and after in vivo recovery was subsequently determined by MTT assay. Additionally, IB structures were measured by Fourier transform infrared (FT-IR) spectroscopy and fluorescence spectroscopy. The results showed that after in vivo refolding, IBs retained lower levels of anti-tumor activity and fewer native-like β-sheet structures. Fewer recoverable polypeptides were trapped in IBs after GroEL/ES co-expression and refolding in vivo. Therefore, GroEL/ES mediated the in vivo recovery of TRAIL IBs in Escherichia coli. These results may identify potential uses for IBs and provide additional insight into the detailed mechanisms of in vivo protein recovery.Zhanqing WangMin ZhangXin LvJiying FanJian ZhangJing SunYaling ShenNature PortfolioarticleTumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL)Soluble TRAILrhTRAILGroE ChaperoninColi Strain C600MedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-10 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) Soluble TRAIL rhTRAIL GroE Chaperonin Coli Strain C600 Medicine R Science Q
spellingShingle	Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) Soluble TRAIL rhTRAIL GroE Chaperonin Coli Strain C600 Medicine R Science Q Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
description	Abstract Inclusion body (IB) formation generates substantial bio-waste in the pharmaceutical industry and remains a major challenge for heterologous protein expression. Although chaperones can be co-expressed to improve soluble protein yield, their contribution to IB processing in vivo has not been thoroughly studied. Here, a GroEL-GroES co-expressing strain and a deficient strain were constructed to study the in vivo recovery of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). The interaction between GroEL/ES and TRAIL was simulated by molecular docking and identified by co-immunoprecipitation. The in vitro cytotoxicity of TRAIL IBs before and after in vivo recovery was subsequently determined by MTT assay. Additionally, IB structures were measured by Fourier transform infrared (FT-IR) spectroscopy and fluorescence spectroscopy. The results showed that after in vivo refolding, IBs retained lower levels of anti-tumor activity and fewer native-like β-sheet structures. Fewer recoverable polypeptides were trapped in IBs after GroEL/ES co-expression and refolding in vivo. Therefore, GroEL/ES mediated the in vivo recovery of TRAIL IBs in Escherichia coli. These results may identify potential uses for IBs and provide additional insight into the detailed mechanisms of in vivo protein recovery.
format	article
author	Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen
author_facet	Zhanqing Wang Min Zhang Xin Lv Jiying Fan Jian Zhang Jing Sun Yaling Shen
author_sort	Zhanqing Wang
title	GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
title_short	GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
title_full	GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
title_fullStr	GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
title_full_unstemmed	GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli
title_sort	groel/es mediated the in vivo recovery of trail inclusion bodies in escherichia coli
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/9c001b387a9d4afc83e57e681db6a0bc
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_version_	1718388015886237696

GroEL/ES mediated the in vivo recovery of TRAIL inclusion bodies in Escherichia coli

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