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...
| Published in: | E3S Web of Conferences |
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| Main Authors: | , |
| Format: | Article in Journal/Newspaper |
| Language: | English French |
| Published: |
EDP Sciences
2020
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| Subjects: | |
| Online Access: | https://doi.org/10.1051/e3sconf/202018504050 https://doaj.org/article/976d931c2d424142bce8a47f2dd672e0 |
| Summary: | 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. |
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