The energy-coupling controlled efflux of 2-keto-3-deoxy-D-gluconate in Escherichia coli K 12. 1975

A E Lagarde, and F R Stoeber

Experiments were devised to test the plausibility and the predictions of a efflux rate equation which was previously derived [10]9 1. 2-Keto-3-deoxy-D-gluconate transport system conforms with universal laws relating zero-trans influx, influx at steady-state, steady-state levels of accumulation to external and internal substrate concentrations. 2. Full-time-course uptake kinetics fit the linearized graphical representation that can be inferred from the integrated rate equation. 3. Influx does not depend upon internal substrate concentration nor upon energy-coupling. 4. Zero-trans outflux (leak inot empty medium) is a first-order process (rate constant: 0.02 min-1) and not mediated by the carrier. Absence of cis-competition with D-glucuronate is in agreement with a simple diffusion mechanism. 5. Outflux increases when external substrate concentration is raised (counterflow). Outflux at steady-state equilibrates influx, and is a first-order process (rate constant: 0.15 min-1). 6. Uncoupling with azide leads to accelerate zero-trans outflux by a factor of 2-3. No further acceleration is obtained when other classical uncouplers are used. The process remains first-order, independent of the amount of carrier, and is accelerated by the presence of internal D-glucuronate as a result from trans-inhibition of the recapture. 7. Exchange outflux is all the more accelerated by azide as the carrier is less saturated. The process is clearly carrier-mediated and the outflux rate obeys a Michaelis law with respect to internal concentration. V is equal to V for influx. 8. Homo and hetero-overshoot experiments are in agreement with the participation of the carrier for mediating influx as well as outflux. 9. The kinetics of D-glucuronate outflux in a strain lacking the specific hexuronate permease but carrying the 2-keto-3-deoxy-D-glucuronate permease are similar to those obtained with 2-keto-3-deoxy-D-gluconate. We draw the conclusion that energy-coupling promotes the adjustment of outflux without interfering with influx rate. It apparently acts by reducing, in a continuous range, the affinity of the carrier facing inwards. The discussion is focused on the comparison with previously published models and on possible molecular mechanisms.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008433 Mathematics The deductive study of shape, quantity, and dependence. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Mathematic
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005942 Gluconates Derivatives of gluconic acid (the structural formula HOCH2(CHOH)4COOH), including its salts and esters. Copper Gluconate,Gluconate, Copper
D005965 Glucuronates Derivatives of GLUCURONIC ACID. Included under this heading are a broad variety of acid forms, salts, esters, and amides that include the 6-carboxy glucose structure. Glucosiduronates,Glucuronic Acids,Acids, Glucuronic
D001386 Azides Organic or inorganic compounds that contain the -N3 group. Azide
D001692 Biological Transport The movement of materials (including biochemical substances and drugs) through a biological system at the cellular level. The transport can be across cell membranes and epithelial layers. It also can occur within intracellular compartments and extracellular compartments. Transport, Biological,Biologic Transport,Transport, Biologic
D001693 Biological Transport, Active The movement of materials across cell membranes and epithelial layers against an electrochemical gradient, requiring the expenditure of metabolic energy. Active Transport,Uphill Transport,Active Biological Transport,Biologic Transport, Active,Transport, Active Biological,Active Biologic Transport,Transport, Active,Transport, Active Biologic,Transport, Uphill

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