Purification of proteins fused to maltose-binding protein.
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.
Lebendiker
Mario
M
Protein Expression and Purification Facility, The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem, Jerusalem, Israel.
Danieli
Tsafi
T
D000688:Amylose / Q000737:chemistry; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D017743:Equipment Reuse; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D058619:Maltose-Binding Proteins / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000302:isolation & purification*; D012685:Sepharose / Q000737:chemistry
D058619:Maltose-Binding Proteins; D011993:Recombinant Fusion Proteins; D000688:Amylose; D012685:Sepharose