The state of energization of the membrane of Escherichia coli as affected by physiological conditions and colicin K.
The bacterial protein colicin K, when added to sensitive Escherichia coli in the presence of 3,3'-dihexyloxacarbocyanine, cuases a doubling in fluorescence of the probe. Glucose and oxygen cause a decreased fluorescence while anoxia and cyanide cause a rise in fluorescence. These results in conjunction with the work of other laboratories suggest that colicin K causes a depolarization of the transmembrane electrical potential. Fluorescence in the absence of colicin K was relatively independent of KCl, NaCl, and MgCl2 concentrations below 0.1 M. Although colicin K caused rapid efflux of the K+ analogue 86Rb+, the fluorescence rise was only partially blocked by 0.13 M KCl. The level of fluorescence caused by the action of colcin K was inversely proportional to the logarithm of the concentration of MgCl2 over the range of 2 muM to 4 mM. This suggests that a Nernst electrochemical potential for an anion can counteract a membrane depolarization caused by colcin. After colcin K action, the fluorescence of the carbocyanine could be further increased by anoxia or cyanide. The distribution of the weak base dimethyloxazolidinedione indicated that the pH in the interior of aerobic E. coli supplied with lactate was alkaline by 0.1 unit and unaffected by colicin. These results suggest that colicin K does not completely depolarize the membrane potential and does not interfere with the component of membrane energization generated by electron transport. Colicin K does not act as a cationophore. The partial depolarization of the membrane may account for the inhibition of active solute transport caused by colicin K.
D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D003087:Colicins / Q000494:pharmacology*; D003486:Cyanides / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D010710:Phosphates / Q000378:metabolism; D011189:Potassium Chloride / Q000494:pharmacology; D011199:Potentiometry; D012413:Rubidium / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D013050:Spectrometry, Fluorescence
D003087:Colicins; D003486:Cyanides; D010710:Phosphates; D012965:Sodium Chloride; D011189:Potassium Chloride; D008274:Magnesium; D005947:Glucose; D012413:Rubidium