title: The requirement for energy during export of beta-lactamase in Escherichia coli is fulfilled by the total protonmotive force.
abstract: The energy requirement for the maturation and export of the plasmid-encoded TEM beta-lactamase in Escherichia coli K12 was shown to be fulfilled by the total protonmotive force. This was demonstrated by assessing the inhibition of proteolytic processing of the precursor form of beta-lactamase caused by perturbation of the energized state of the membrane in cells treated with valinomycin. The magnitude of the membrane potential was manipulated by varying the concentration of KCl in the medium and the pH gradient was manipulated by varying the external pH. Both components were simultaneously affected by addition of the protonophore carbonylcyanide-p- trifluoromethoxy phenylhydrazone (FCCP). Inhibition of processing was demonstrated in a mutant strain having a defective ATP synthase where protonmotive force could be dissipated without altering the intracellular level of ATP, indicating that the observed inhibition was not the result of decreased ATP concentration. Half-maximal accumulation of precursor of beta-lactamase was observed in all cases when the level of protonmotive force was decreased to approximately 150 mV. Under those conditions the membrane potential varied from 65 to 140 mV (internally negative) and the pH gradient from 95 to 25 mV (internally alkaline). Thus, the energy requirement is satisfied by the total protonmotive force, with no specificity for either the membrane potential or the pH gradient.
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001693:Biological Transport, Active / Q000187:drug effects; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D004563:Electrochemistry; D004734:Energy Metabolism; D004792:Enzyme Precursors / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials; D011189:Potassium Chloride / Q000494:pharmacology; D011522:Protons; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D004792:Enzyme Precursors; D011522:Protons; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D001618:beta-Lactamases