The proton motive force lowers the level of ATP required for the in vitro translocation of a secretory protein in Escherichia coli.
The role of the electrochemical potential difference of proton (delta mu H+) in protein translocation across the membrane of Escherichia coli was examined in detail using an efficient in vitro assay system (Yamada, H., Tokuda, H., and Mizushima, S. (1989) J. Biol. Chem. 264, 1723-1728). Delta mu H+ reduced the level of ATP necessary for the efficient translocation of OmpF-Lpp, a chimeric model secretory protein. The apparent Km value of the translocation reaction for ATP was lower by 2 orders of magnitude in the presence of delta mu H+ than in its absence. The membrane potential and delta pH, both of which are components of delta mu H+, independently lowered the apparent Km value of the translocation reaction for ATP. An ATP-generating system also lowered the level of ATP required for translocation in the absence of delta mu H+ but not in its presence. It is proposed that ADP formed during protein translocation lowers the affinity of the putative translocation machinery for ATP and that the removal of ADP from the secretory machinery, a possible critical step in the translocation reaction, is stimulated in the presence of either delta mu H+, an ATP-generating system, or a higher concentration of ATP.
Shiozuka
K
K
Institute of Applied Microbiology, University of Tokyo, Japan.
Tani
K
K
Mizushima
S
S
Tokuda
H
H
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology; D002678:Chimera; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008564:Membrane Potentials; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins; D014158:Transcription, Genetic
D001425:Bacterial Outer Membrane Proteins; D008074:Lipoproteins; C066293:OmpF-Lpp chimeric protein; D011993:Recombinant Fusion Proteins; D000255:Adenosine Triphosphate