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.
Protein Expression and Purification Facility, The Wolfson Centre for Applied Structural Biology, The Hebrew University of Jerusalem, Jerusalem, Israel.
Mario
M
Lebendiker
Tsafi
T
Danieli
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