The characterization of energized and partially de-energized (respiration-independent) beta-galactoside transport into Escherichia coli.
Unidirectional fluxes of [14C]lactose by whole cells of Escherichia coli under highly energized and partially de-energized (in the presence of CN-) conditions are analyzed kinetically. When the cells are energized, the value for V influx is 0.45 +/- 0.01 mM internal concentration increment/s and Kt is 0.26 +/- 0.03 mM. At an external concentration of 0.61 mM the steady-state internal concentration is 0.25 M, reached after about 1h. The maximum steady-state concentration ratio is 2-10(3). The efflux process under these conditions is non-saturable, being linearly dependent upon internal concentration over the range 25-250 mM with a first-order rate constant of 8.8 +/- 0.2-10(-4) S-1. The transport in the presence of CN- is active, with a maximum concentration ratio (internal concentration/external concentration) of 104, and the uptake is mimicked by anoxia (less than 70 ppm O2). The effects of CN- are to lower the V for influx and to change the efflux from a non-saturable to a saturable process with a value for Kt (60 mM) intermediate between that for energized efflux (greater than 250 mM) and influx (0.3-0.6 mM), the latter value not changing appreciably. Partial de-energization thus affects both the influx and efflux processes.
D000332:Aerobiosis; D000693:Anaerobiosis; D001693:Biological Transport, Active; D002701:Chloramphenicol / Q000494:pharmacology; D003486:Cyanides / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D007785:Lactose / Q000378:metabolism*; D008433:Mathematics; D010101:Oxygen Consumption / Q000187:drug effects
D003486:Cyanides; D002701:Chloramphenicol; D007785:Lactose