Abstract
Maltose-binding protein (MBP) is one of the most popular fusion partners being used for producing recombinant proteins in bacterial cells. MBP allows one to use a simple capture affinity step on amylose–agarose columns, resulting in a protein that is often 70–90% pure. In addition to protein-isolation applications, MBP provides a high degree of translation and facilitates the proper folding and solubility of the target protein. This chapter describes efficient procedures for isolating highly purified MBP-target proteins. Special attention is given to considerations for downstream applications such as structural determination studies, protein activity assays, and assessing the chemical characteristics of the target protein.
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Acknowledgments
We thank Dr. Erez Podoly from the Biological Chemistry Department of The Hebrew University of Jerusalem, now at the Structural Biology Department of Stanford University School of Medicine, for expressing the fusion protein, for his assistance with purification and his helpful suggestions. We thank Dr. Daria Mochly-Rosen from the Department of Chemical and Systems Biology, School of Medicine, Stanford University for kindly providing us with pMAL-c2MBP-Rack1.
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Lebendiker, M., Danieli, T. (2011). Purification of Proteins Fused to Maltose-Binding Protein. In: Walls, D., Loughran, S. (eds) Protein Chromatography. Methods in Molecular Biology, vol 681. Humana Press. https://doi.org/10.1007/978-1-60761-913-0_15
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DOI: https://doi.org/10.1007/978-1-60761-913-0_15
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