Sodium ion-dependent amino acid transport in membrane vesicles of Bacillus stearothermophilus.
Amino acid transport in membrane vesicles of Bacillus stearothermophilus was studied. A relatively high concentration of sodium ions is needed for uptake of L-alanine (Kt = 1.0 mM) and L-leucine (Kt = 0.4 mM). In contrast, the Na(+)-H(+)-L-glutamate transport system has a high affinity for sodium ions (Kt less than 5.5 microM). Lithium ions, but no other cations tested, can replace sodium ions in neutral amino acid transport. The stimulatory effect of monensin on the steady-state accumulation level of these amino acids and the absence of transport in the presence of nonactin indicate that these amino acids are translocated by a Na+ symport mechanism. This is confirmed by the observation that an artificial delta psi and delta mu Na+/F but not a delta pH can act as a driving force for uptake. The transport system for L-alanine is rather specific. L-Serine, but not L-glycine or other amino acids tested, was found to be a competitive inhibitor of L-alanine uptake. On the other hand, the transport carrier for L-leucine also translocates the amino acids L-isoleucine and L-valine. The initial rates of L-glutamate and L-alanine uptake are strongly dependent on the medium pH. The uptake rates of both amino acids are highest at low external pH (5.5 to 6.0) and decline with increasing pH. The pH allosterically affects the L-glutamate and L-alanine transport systems. The maximal rate of L-glutamate uptake (Vmax) is independent of the external pH between pH 5.5 and 8.5, whereas the affinity constant (Kt) increases with increasing pH. A specific transport system for the basic amino acids L-lysine and L-arginine in the membrane vesicles has also been observed. Transport of these amino acids occurs most likely by a uniport mechanism.
Heyne
R I
RI
Department of Microbiology, University of Groningen, Haren, The Netherlands.
de Vrij
W
W
Crielaard
W
W
Konings
W N
WN
D000409:Alanine / Q000378:metabolism; D000495:Allosteric Site; D000596:Amino Acids / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001667:Binding, Competitive; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D001411:Geobacillus stearothermophilus / Q000378:metabolism*; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D006863:Hydrogen-Ion Concentration; D007476:Ionophores / Q000494:pharmacology; D007700:Kinetics; D007930:Leucine / Q000494:pharmacology; D008239:Lysine / Q000378:metabolism; D018942:Macrolides; D008985:Monensin / Q000494:pharmacology; D012964:Sodium / Q000494:pharmacology*
D000596:Amino Acids; D000900:Anti-Bacterial Agents; D005971:Glutamates; D007476:Ionophores; D018942:Macrolides; D018698:Glutamic Acid; D008985:Monensin; D012964:Sodium; D007930:Leucine; D008239:Lysine; D000409:Alanine; C050476:nonactin