Campus Units
Biochemistry, Biophysics and Molecular Biology, Roy J. Carver Department of
Document Type
Article
Publication Version
Accepted Manuscript
Publication Date
8-2008
Journal or Book Title
Protein Expression and Purification
Volume
60
Issue
2
First Page
194
Last Page
197
DOI
10.1016/j.pep.2008.04.001
Abstract
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 copurifying 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.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Copyright Owner
Elsevier Inc.
Copyright Date
2008
Language
en
File Format
application/pdf
Recommended Citation
Joseph, Raji E. and Andreotti, Amy H., "Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity" (2008). Biochemistry, Biophysics and Molecular Biology Publications. 190.
https://lib.dr.iastate.edu/bbmb_ag_pubs/190
Included in
Biochemistry Commons, Biophysics Commons, Molecular Biology Commons, Structural Biology Commons
Comments
This is a manuscript of an article published as Joseph, Raji E., and Amy H. Andreotti. "Bacterial expression and purification of interleukin-2 tyrosine kinase: single step separation of the chaperonin impurity." Protein expression and purification 60, no. 2 (2008): 194-197. doi: 10.1016/j.pep.2008.04.001. Posted with permission.