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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Bibliographic Details
Main Authors: Joseph, Raji E., Andreotti, Amy H.
Format: Text
Language:English
Published: Iowa State University Digital Repository 2008
Subjects:
kinase
bactieral expression
purification
chaperonin
Itk
Lck
Src Introduction
Biochemistry
Biophysics
Molecular Biology
Structural Biology
Antarc*
Antarctica
Online Access:https://lib.dr.iastate.edu/bbmb_ag_pubs/190
https://lib.dr.iastate.edu/cgi/viewcontent.cgi?article=1196&context=bbmb_ag_pubs
Description
Summary: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.

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