Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli
Cytosolic carboxypeptidase 6 (CCP6) is a member of cytosolic carboxypeptidase (CCP) family that catalyze the removal of polyglutamate side chains from protein substrates. Biochemical and biophysical characterization of CCPs requires large quantities of purified proteins. However, no method describin...
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ftdoajarticles:oai:doaj.org/article:976d931c2d424142bce8a47f2dd672e0 2023-05-15T13:57:29+02:00 Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli Wang Ruixue Wu Hui-Yuan 2020-01-01T00:00:00Z https://doi.org/10.1051/e3sconf/202018504050 https://doaj.org/article/976d931c2d424142bce8a47f2dd672e0 EN FR eng fre EDP Sciences https://www.e3s-conferences.org/articles/e3sconf/pdf/2020/45/e3sconf_iceeb2020_04050.pdf https://doaj.org/toc/2267-1242 2267-1242 doi:10.1051/e3sconf/202018504050 https://doaj.org/article/976d931c2d424142bce8a47f2dd672e0 E3S Web of Conferences, Vol 185, p 04050 (2020) Environmental sciences GE1-350 article 2020 ftdoajarticles https://doi.org/10.1051/e3sconf/202018504050 2022-12-31T12:18:24Z Cytosolic carboxypeptidase 6 (CCP6) is a member of cytosolic carboxypeptidase (CCP) family that catalyze the removal of polyglutamate side chains from protein substrates. Biochemical and biophysical characterization of CCPs requires large quantities of purified proteins. However, no method describing the expression of any mammalian CCP family member from bacteria has been published to our best knowledge. After considerable efforts to improve the solubility of mammalian CCPs expressed in bacteria, including the optimization of induction temperature and by using different receptive cells, we were able to get less expression of mouse CCP6 in soluble fraction of bacterial lysates. We report in this article, the bacterial expression of CCP6 using Arctic Express (DE3) competent cells that co-express the chaperonin system GroEL and GroES from Oleispira antarctica. However, to achieve a large number of soluble target proteins, the expression conditions still need to be further optimized. Article in Journal/Newspaper Antarc* Antarctica Arctic Directory of Open Access Journals: DOAJ Articles Arctic E3S Web of Conferences 185 04050 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English French |
topic |
Environmental sciences GE1-350 |
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Environmental sciences GE1-350 Wang Ruixue Wu Hui-Yuan Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
topic_facet |
Environmental sciences GE1-350 |
description |
Cytosolic carboxypeptidase 6 (CCP6) is a member of cytosolic carboxypeptidase (CCP) family that catalyze the removal of polyglutamate side chains from protein substrates. Biochemical and biophysical characterization of CCPs requires large quantities of purified proteins. However, no method describing the expression of any mammalian CCP family member from bacteria has been published to our best knowledge. After considerable efforts to improve the solubility of mammalian CCPs expressed in bacteria, including the optimization of induction temperature and by using different receptive cells, we were able to get less expression of mouse CCP6 in soluble fraction of bacterial lysates. We report in this article, the bacterial expression of CCP6 using Arctic Express (DE3) competent cells that co-express the chaperonin system GroEL and GroES from Oleispira antarctica. However, to achieve a large number of soluble target proteins, the expression conditions still need to be further optimized. |
format |
Article in Journal/Newspaper |
author |
Wang Ruixue Wu Hui-Yuan |
author_facet |
Wang Ruixue Wu Hui-Yuan |
author_sort |
Wang Ruixue |
title |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
title_short |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
title_full |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
title_fullStr |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
title_full_unstemmed |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli |
title_sort |
expression of recombinant mouse cytosolic carboxypeptidase 6 in escherichia coli |
publisher |
EDP Sciences |
publishDate |
2020 |
url |
https://doi.org/10.1051/e3sconf/202018504050 https://doaj.org/article/976d931c2d424142bce8a47f2dd672e0 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Antarc* Antarctica Arctic |
genre_facet |
Antarc* Antarctica Arctic |
op_source |
E3S Web of Conferences, Vol 185, p 04050 (2020) |
op_relation |
https://www.e3s-conferences.org/articles/e3sconf/pdf/2020/45/e3sconf_iceeb2020_04050.pdf https://doaj.org/toc/2267-1242 2267-1242 doi:10.1051/e3sconf/202018504050 https://doaj.org/article/976d931c2d424142bce8a47f2dd672e0 |
op_doi |
https://doi.org/10.1051/e3sconf/202018504050 |
container_title |
E3S Web of Conferences |
container_volume |
185 |
container_start_page |
04050 |
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1766265155609427968 |