Glutamate transport in Escherichia coli K-12: nonidentity of carriers mediating entry and exit.
The exit of glutamate from Escherichia coli K-12 cells preloaded with the radioactive amino acid and its relation to the reaction of entry were studied. Experiments with cells preloaded to different intracellular concentrations of radioactive glutamate confirmed our earlier conclusion that glutamate exit was a first-order reaction. l-Glutamate, competitive inhibitors of glutamate uptake (d-glutamate and l-glutamate-gamma-methyl ester), noncompetitive inhibitors of glutamate uptake (l-serine and l-alanine), and the energy poison NaN(3) all accelerated glutamate exit 2.8-fold. No additive effect was observed in the presence of NaN(3) together with l-glutamate. Preloading with cold l-glutamate did not increase the rate of uptake of radioactive glutamate. It is concluded that the acceleration of glutamate exit in the presence of l-glutamate in the medium is not due to exchange diffusion and that l-glutamate and azide affect exit indirectly by preventing recapture. Sucrose, 25%, slowed down glutamate exit by a factor of about 4.7 and increased the steady-state level of glutamate accumulation to about the same extent. Increasing the intracellular K(+) concentration enhanced glutamate uptake but did not affect the half-time of exit. It is concluded that separate carriers are most probably involved in mediating the entry and exit reactions.
D000409:Alanine / Q000494:pharmacology; D001386:Azides / Q000494:pharmacology; D001693:Biological Transport, Active; D002247:Carbon Isotopes; D003470:Culture Media; D004058:Diffusion; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D004952:Esters / Q000494:pharmacology; D005971:Glutamates / Q000378:metabolism* / Q000494:pharmacology; D007930:Leucine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D012694:Serine / Q000494:pharmacology; D013237:Stereoisomerism; D013395:Sucrose / Q000494:pharmacology
D001386:Azides; D002247:Carbon Isotopes; D003470:Culture Media; D004952:Esters; D005971:Glutamates; D012694:Serine; D013395:Sucrose; D008715:Methionine; D007930:Leucine; D000409:Alanine