Mechanism and energetics of dipeptide transport in membrane vesicles of Lactococcus lactis.
Alanyl-alpha-glutamate transport has been studied in Lactococcus lactis ML3 cells and in membrane vesicles fused with liposomes containing beefheart cytochrome c oxidase as a proton-motive-force-generating system. The uptake of Ala-Glu observed in de-energized cells can be stimulated 26-fold upon addition of lactose. No intracellular dipeptide pool could be detected in intact cells. In fused membranes, a 40-fold accumulation of Ala-Glu was observed in response to a proton motive force. Addition of ionophores and uncouplers resulted in a rapid efflux of the accumulated dipeptide, indicating that Ala-Glu accumulation is directly coupled to the proton motive force as a driving force. Ala-Glu uptake is an electrogenic process and the dipeptide is transported in symport with two protons. In both fused membranes and intact cells the same affinity constant (0.70 mM) for Ala-Glu uptake was found. Accumulated Ala-Glu is exchangeable with externally added alanyl-glutamate, glutamyl-glutamate, and leucyl-leucine, while no exchange occurred upon addition of the amino acid glutamate or alanine. These results indicate that the Ala-Glu transport system has a broad substrate specificity.
Smid
E J
EJ
Department of Microbiology, University of Groningen, Haren, The Netherlands.
Driessen
A J
AJ
Konings
W N
WN
D000596:Amino Acids / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology; D004151:Dipeptides / Q000378:metabolism*; D005971:Glutamates / Q000378:metabolism; D007700:Kinetics; D013294:Lactococcus lactis / Q000378:metabolism*; D008081:Liposomes; D008564:Membrane Potentials; D009550:Nigericin / Q000494:pharmacology; D014634:Valinomycin / Q000494:pharmacology
D000596:Amino Acids; D004151:Dipeptides; D005971:Glutamates; D008081:Liposomes; D014634:Valinomycin; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; C045560:alanylglutamic acid; D009550:Nigericin