Models to determine the kinetic mechanisms of ion-coupled transporters.

With high-resolution structures available for many ion-coupled (secondary active) transporters, a major challenge for the field is to determine how coupling is accomplished. Knowledge of the kinetic mechanism of the transport reaction, which defines the binding order of substrate and co-ions, together with the sequence with which all relevant states are visited by the transporter, will help to reveal this coupling mechanism. Here, we derived general mathematical models that can be used to analyze data from steady-state transport measurements and show how kinetic mechanisms can be derived.

Arginine and Citrulline Catabolic Pathways Encoded by the Gene Cluster of Lactobacillus brevis ATCC 367.

High concentrations of l-arginine or l-citrulline in the growth medium provided the wine bacterium with a significant growth advantage. The arginine deiminase pathway (ADI) gene cluster of contains three genes-, , and -encoding putative l-arginine/l-ornithine exchangers. Uptake experiments with cells expressing the genes showed that all three transported l-ornithine with affinities in the micromolar range.

Convergent evolution of the arginine deiminase pathway: the ArcD and ArcE arginine/ornithine exchangers.

The arginine deiminase (ADI) pathway converts L-arginine into L-ornithine and yields 1 mol of ATP per mol of L-arginine consumed. The L-arginine/L-ornithine exchanger in the pathway takes up L-arginine and excretes L-ornithine from the cytoplasm. Analysis of the genomes of 1281 bacterial species revealed the presence of 124 arc gene clusters encoding the pathway.

Structure and elevator mechanism of the Na-citrate transporter CitS.

The recently determined crystal structure of the bacterial Na-citrate symporter CitS provides unexpected structural and mechanistic insights. The protein has a fold that has not been seen in other proteins, but the oligomeric state, domain organization and proposed transport mechanism strongly resemble those of the sodium-dicarboxylate symporter vcINDY, and the putative exporters YdaH and MtrF, thus hinting at convergence in structure and function. CitS and the related proteins are predicted to translocate their substrates by an elevator-like mechanism, in which a compact transport domain slides up and down through the membrane while the dimerization domain is stably anchored.

ArcD1 and ArcD2 Arginine/Ornithine Exchangers Encoded in the Arginine Deiminase Pathway Gene Cluster of Lactococcus lactis.

Unlabelled: The arginine deiminase (ADI) pathway gene cluster in Lactococcus lactis contains two copies of a gene encoding an l-arginine/l-ornithine exchanger, the arcD1 and arcD2 genes. The physiological function of ArcD1 and ArcD2 was studied by deleting the two genes. Deletion of arcD1 resulted in loss of the growth advantage observed in the presence of high l-arginine in different growth media.