Escherichia coli lacking the AcrAB multidrug efflux pump also lacks nonproteinaceous, PHB-polyphosphate Ca2+ channels in the membrane.
PHB(polyP) complexes bind calcium and form calcium channels in the cytoplasmic membrane in Escherichia coli and are likely to be important in Ca(2+) homeostasis in this organism. E. coli N43, which lacks the AcrA component of a major multidrug resistance pump, was shown to be defective in calcium handling, with an inability to maintain submicromolar levels of free Ca(2+) in the cytoplasm. Therefore, using an N-phenyl-1-napthylamine (NPN)-dependent fluorescence assay, we measured temperature-dependent phase transitions in the membranes of intact cells. These transitions specifically depend on the presence of PHB(Ca(2+)polyP) complexes. PHB(Ca(2+)polyP) channel complexes, particularly in stationary phase cultures, were detected in wild-type strains; however, in contrast, isogenic acrA(-) strains had greatly reduced amounts of the complexes. This indicates that the AcrAB transporter may have a novel, hitherto undetected physiological role, either directly in the membrane assembly of the PHB complexes or the transport of a component of the membrane, which is essential for assembly of the complexes into the membrane. In other experiments, we showed that the particular defective calcium handling detected in N43 was not due to the absence of AcrA but to other unknown factors in this strain.
Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, UK.
Helen E
HE
Jones
I Barry
IB
Holland
Annicq
A
Jacq
Torun
T
Wall
Anthony K
AK
Campbell
D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000502:physiology*; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology*; D006885:Hydroxybutyrates / Q000032:analysis*; D008074:Lipoproteins / Q000235:genetics / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011091:Polyesters / Q000032:analysis*; D000094122:Polyhydroxybutyrates
D002118:Calcium; D015220:Calcium Channels; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D006885:Hydroxybutyrates; D008074:Lipoproteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D011091:Polyesters; C083497:AcrA protein, E coli; C083496:AcrB protein, E coli; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates