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Article
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity
Protein Expression and Purification
  • Raji E. Joseph, Iowa State University
  • Amy H. Andreotti, Iowa State University
Document Type
Article
Publication Version
Accepted Manuscript
Publication Date
8-1-2008
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.

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.

Creative Commons License
Creative Commons Attribution-Noncommercial-No Derivative Works 4.0
Copyright Owner
Elsevier Inc.
Language
en
File Format
application/pdf
Citation Information
Raji E. Joseph and Amy H. Andreotti. "Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity" Protein Expression and Purification Vol. 60 Iss. 2 (2008) p. 194 - 197
Available at: http://works.bepress.com/amy_andreotti/20/