Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity
Biochemical and biophysical characterization of kinases requires large quantities of purified protein. Here we report the bacterial expression and purification of active Itk kinase domain (a Tec family kinase) using ArcticExpress cells that co-express the chaperonin system Cpn60/10 from Oleispira an...
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Online Access: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581883 http://www.ncbi.nlm.nih.gov/pubmed/18495488 https://doi.org/10.1016/j.pep.2008.04.001 |
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ftpubmed:oai:pubmedcentral.nih.gov:2581883 2023-05-15T13:35:32+02:00 Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity Joseph, Raji E. Andreotti, Amy H. 2008-04-11 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581883 http://www.ncbi.nlm.nih.gov/pubmed/18495488 https://doi.org/10.1016/j.pep.2008.04.001 en eng http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581883 http://www.ncbi.nlm.nih.gov/pubmed/18495488 http://dx.doi.org/10.1016/j.pep.2008.04.001 Article Text 2008 ftpubmed https://doi.org/10.1016/j.pep.2008.04.001 2013-09-02T07:44:23Z Biochemical and biophysical characterization of kinases requires large quantities of purified protein. Here we report the bacterial expression and purification of active Itk kinase domain (a Tec family kinase) using ArcticExpress cells that co-express the chaperonin system Cpn60/10 from Oleispira antarctica. We describe a simple one step MgCl2/ATP/KCl incubation procedure to remove the co-purifying chaperonin impurity. Chaperonin co-purification is a common problem encountered during protein purification and the simple incubation step described here completely overcomes this problem. The approach targets the chaperonin system rather than the protein of interest and is therefore widely applicable to other protein targets. Text Antarc* Antarctica PubMed Central (PMC) Protein Expression and Purification 60 2 194 197 |
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Article Joseph, Raji E. Andreotti, Amy H. Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
topic_facet |
Article |
description |
Biochemical and biophysical characterization of kinases requires large quantities of purified protein. Here we report the bacterial expression and purification of active Itk kinase domain (a Tec family kinase) using ArcticExpress cells that co-express the chaperonin system Cpn60/10 from Oleispira antarctica. We describe a simple one step MgCl2/ATP/KCl incubation procedure to remove the co-purifying chaperonin impurity. Chaperonin co-purification is a common problem encountered during protein purification and the simple incubation step described here completely overcomes this problem. The approach targets the chaperonin system rather than the protein of interest and is therefore widely applicable to other protein targets. |
format |
Text |
author |
Joseph, Raji E. Andreotti, Amy H. |
author_facet |
Joseph, Raji E. Andreotti, Amy H. |
author_sort |
Joseph, Raji E. |
title |
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
title_short |
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
title_full |
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
title_fullStr |
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
title_full_unstemmed |
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity |
title_sort |
bacterial expression and purification of interleukin-2 tyrosine kinase: single step separation of the chaperonin impurity |
publishDate |
2008 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581883 http://www.ncbi.nlm.nih.gov/pubmed/18495488 https://doi.org/10.1016/j.pep.2008.04.001 |
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Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2581883 http://www.ncbi.nlm.nih.gov/pubmed/18495488 http://dx.doi.org/10.1016/j.pep.2008.04.001 |
op_doi |
https://doi.org/10.1016/j.pep.2008.04.001 |
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Protein Expression and Purification |
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60 |
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2 |
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194 |
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197 |
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1766066795793350656 |