PMID:8954986
| Citation |
Van Gelder, P, Steiert, M, El Khattabi, M, Rosenbusch, JP and Tommassen, J (1996) Structural and functional characterization of a His-tagged PhoE pore protein of Escherichia coli. Biochem. Biophys. Res. Commun. 229:869-75 |
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| Abstract |
The recent elucidation of the 3-D structure of the outer membrane protein PhoE of Escherichia coli provides an excellent tool for a detailed analysis of the structure-function relationship of this pore-forming protein. For this purpose, a fast and efficient method for the purification of mutant porins is needed. A histidine-tag was engineered between the signal sequence and the N terminus of mature PhoE. The recombinant PhoE protein was normally assembled into the outer membrane and could be purified by immobilized metal affinity chromatography. Part of the total amount of the trimers dissociated into folded monomers during purification. The histidine-tag did not change the electrophysical characteristics of the protein in lipid bilayers. Hence, the method is useful for the fast purification of mutant porins for functional and structural characterization. |
| Links | |
| Keywords |
Escherichia coli/metabolism; Escherichia coli Proteins; Histidine; Porins/chemistry; Porins/isolation & purification; Porins/metabolism; Structure-Activity Relationship |
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