Abstract
Efficient translocation of pure precursor of PhoE protein (prePhoE) could be accomplished in an in vitro system consisting of only inverted Escherichia coli inner membrane vesicles, ATP, and SecA and SecB protein. In this in vitro system SecB and not trigger factor could stabilize a translocation-competent state of prePhoE. In contrast, translocation competency of proOmpA could be induced by both trigger factor and SecB protein, suggesting specificity in interactions between cytosolic factors and precursors in outer membrane protein translocation.
Highlights
Ron KustersS, Truus de VrijeSg, Eefjan BreukinkS, in uitro-synthesized precursor of the E . coli outer membrane and Ben de KruijffSP pore protein PhoEcross inner membranevesicles
Translocation competency of proOmpA could be induced by both trigger factor and
The precursor could be stabilized for translocation monoQ column (Pharmacia, Sweden) in 8 M urea, 10 mM Tris/HCl, by stoichiometricinteractionwith trigger factor (8).By means of the asoluble protein called same reconstituted system pH 9.0
Summary
Efficient translocation of pure precursor of PhoE purification of prePhoE which can be effectively translocated protein (prePhoE) could be accomplished in an in vitro systemconsisting of only inverted Escherichiacoli inner membrane vesicles, ATP, and SecA and SecB protein. In this in vitro system SecB and not trigger factor could stabilize a translocation-competent state into inverted membranevesicles in the presence of only ATP and SecA and SecB protein. The precursor could be stabilized for translocation monoQ column (Pharmacia, Sweden) in 8 M urea, 10 mM Tris/HCl, by stoichiometricinteractionwith trigger factor (8).By means of the asoluble protein called same reconstituted system pH 9.0.
Sign-in/Register to view highlights & summary 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 call
