Abstract
The recent elucidation of the 3-D structure of the outer membrane protein PhoE ofEscherichia coliprovides 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.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Biochemical and Biophysical Research Communications
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
