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.

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 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Copyright Owner

Elsevier Inc.

Language

en

File Format

application/pdf

Published Version

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