Energy coupling to active transport in anaerobically grown mutants of Escherichia Coli K12

Abstract

1. Anaerobic uptake of proline requires either the presence of a coupled Mg2+-stimulated adenosine triphosphatase or anaerobic electron transport. 2. Anaerobic uptake of glutamine does not require anaerobic electron transport even in the absence of a coupled Mg+2-stimulated adenosine triphosphatase. 3. These results support previous suggestions [Berger (1973) Proc. Natl. Acad. Sci. U.S.A. 70, 1514––1518; Berger & Heppel (1974) J. Biol. Chem. 249, 7747-7755; Kobayashi, Kin & Anraku (1974) J. Biochem. (Tokyo) 76, 251-261] that two distinct mechanisms of energy coupling to active transport exist in Escherichia coli in that energization of anaerobic proline uptake requires the ‘high-energy membrane state’, whereas the energization of anaerobic glutamine uptake does not.

Uptake of ferrienterochelin by Escherichia coli: energy dependent stage of uptake

“…The effects of DNP and cyanide on glutamine and proline uptake were clearly dependent on the dose of inhibitor used ( Table 2), but we were unable to demonstrate preferential inhibition of proline uptake by the uncoupler CCP ( Table 2). This effect has been noted previously (14) and is thought to be due to the ability of CCP to form thiol groups (22). The uptake of proline was also strongly inhibited by 10 mM sodium azide, which inhibits oxidative phosphorylation and cytochrome oxidase, whereas glutamine uptake was largely unaffected (Fig.…”

“…Effect of anaerobiosis on uptake. Growth of E. coli under anaerobic conditions in the absence of an electron acceptor eliminates electron transport and inhibits respiration-dependent uptake of transport substrates (14). Control experiments demonstrated that glutamine uptake in strain AN248 (unc+) was unaffected by anaerobic growth under iron stress, whereas proline uptake was severely inhibited (20 to.…”

“…The group translocation system is the best understood of these three mechanisms but is applicable only to the uptake of a limited number of sugars (15,38). The remaining two mechanisms can be distinguished by such parameters as sensitivity to uncouplers of oxidative phosphorylation and inhibitors of electron transport or glycolysis, by the effects of loss, through mutation, of cytochrome or adenosine triphosphatase (ATPase) activities, by the direct utilization of fermentable or oxidizable carbon sources to drive active uptake, and by ability to accumulate the transport substrate after anaerobic growth (4,14,23,33). On the basis of results obtained in experiments of this type, several workers postulated that the two energy coupling processes are (i) utilization of an energized membrane state (a proton or charge gradient across the cytoplasmic membrane) generated either by electron transport during respiration or by hydrolysis of adenosine 5'-triphosphate (ATP) by the ATPase under anaerobic conditions, and (ii) direct utilization of a high-energy phosphate bond (probably ATP) (4,14,23).…”

Proline porters effect the utilization of proline as nutrient or osmoprotectant for bacteria

“…The relative cytoplasmic concentrations of proline, glycine betaine, or other solutes maintained by those systems under various conditions have not yet been assessed. Gutowski and Rosenberg (1976) showed that the relative activities of ion symporters and of transporters not energized via the protonmotive force differed under aerobic and anaerobic conditions. Thus the relative contributions of proline porter II, which is energized via respiration, and proline porter III, which is not likely to be energized in that manner, to the accumulation of osmoprotective solutes may be different under aerobic and anaerobic conditions.…”

Pathways of Energy Metabolism Required for Phenotypic Expression of Nif+Kp Genes in Escherichia Coli

“…This requirement can be satisfied either by a functional proton-translocating respiratory chain (with O 2 , fumarate or nitrate as terminal acceptor) (Konings and Kaback 1973;Boonstra et al 1978) or by ATP hydrolysis via the proton-translocating ATPase (Gutowski and Rosenberg 1976;Haddock 1977) (Fig. 1).…”