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Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector

At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chi...

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Detalles Bibliográficos
Autores principales: Li, Jianghui, Han, Qinxia, Zhang, Tao, Du, Jing, Sun, Qianqian, Pang, Yilin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042314/
https://www.ncbi.nlm.nih.gov/pubmed/30009138
http://dx.doi.org/10.1016/j.btre.2018.e00261
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author Li, Jianghui
Han, Qinxia
Zhang, Tao
Du, Jing
Sun, Qianqian
Pang, Yilin
author_facet Li, Jianghui
Han, Qinxia
Zhang, Tao
Du, Jing
Sun, Qianqian
Pang, Yilin
author_sort Li, Jianghui
collection PubMed
description At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chimeric cysteine desulfurase namely, EH-IscS were successfully expressed in E. coli. The proteins were particularly difficult to be produced functionally, due to their readily sequestered nature. The recombinant cysteine desulfurases that were generated by pCold-SUMOa exhibited higher activity, solubility and stability compared with the well-known plasmid pCold I. In contrast to the pCold TF plasmid, the SUMO tag conferred no biological activity with regard to the conformation of the cysteine desulfurases. Furthermore, the SUMO protease 1 can efficiently recognize the tertiary structure of SUMO and cleave it. The data indicate that the pCold-SUMOa vector is a promising tool for native eukaryotic protein production.
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spelling pubmed-60423142018-07-13 Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector Li, Jianghui Han, Qinxia Zhang, Tao Du, Jing Sun, Qianqian Pang, Yilin Biotechnol Rep (Amst) Article At present, approximately 30% of eukaryotic proteins can be expressed in a soluble form in Escherichia coli. In this study, a pCold-SUMOa plasmid was constructed in order to express heterologous proteins fused with SUMO by a cold-shock expression vector. The human cysteine desulfurase NFS1 and a chimeric cysteine desulfurase namely, EH-IscS were successfully expressed in E. coli. The proteins were particularly difficult to be produced functionally, due to their readily sequestered nature. The recombinant cysteine desulfurases that were generated by pCold-SUMOa exhibited higher activity, solubility and stability compared with the well-known plasmid pCold I. In contrast to the pCold TF plasmid, the SUMO tag conferred no biological activity with regard to the conformation of the cysteine desulfurases. Furthermore, the SUMO protease 1 can efficiently recognize the tertiary structure of SUMO and cleave it. The data indicate that the pCold-SUMOa vector is a promising tool for native eukaryotic protein production. Elsevier 2018-05-30 /pmc/articles/PMC6042314/ /pubmed/30009138 http://dx.doi.org/10.1016/j.btre.2018.e00261 Text en © 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Li, Jianghui
Han, Qinxia
Zhang, Tao
Du, Jing
Sun, Qianqian
Pang, Yilin
Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title_full Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title_fullStr Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title_full_unstemmed Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title_short Expression of soluble native protein in Escherichia coli using a cold-shock SUMO tag-fused expression vector
title_sort expression of soluble native protein in escherichia coli using a cold-shock sumo tag-fused expression vector
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6042314/
https://www.ncbi.nlm.nih.gov/pubmed/30009138
http://dx.doi.org/10.1016/j.btre.2018.e00261
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