New substrates for the dicarboxylate transport system of Sinorhizobium meliloti.
The dicarboxylate transport (Dct) system of Sinorhizobium meliloti, which is essential for a functional nitrogen-fixing symbiosis, has been thought to transport only dicarboxylic acids. We show here that the permease component of the Dct system, DctA, can transport orotate, a monocarboxylic acid, with an apparent K(m) of 1.7 mM and a V(max) of 163 nmol min(-1) per mg of protein in induced cells. DctA was not induced by the presence of orotate. The transport of orotate was inhibited by several compounds, including succinamic acid and succinamide, which are not dicarboxylic acids. The dicarboxylic acid maleate (cis-butenedioic acid) was not an inhibitor of orotate transport, which suggests that it was not recognized by DctA. However, maleate was an excellent inducer of DctA expression. Our evaluation of 17 compounds as inducers and inhibitors of transport suggests that substrates recognized by S. meliloti DctA must have appropriately spaced carbonyl groups and an extended conformation, while good inducers are more likely to have a curved conformation.
Yurgel
S
S
Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.
Mortimer
M W
MW
Rogers
K N
KN
Kahn
M L
ML
D001426:Bacterial Proteins / Q000378:metabolism*; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000378:metabolism*; D029382:Dicarboxylic Acid Transporters; D003998:Dicarboxylic Acids / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D009963:Orotic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D016962:Sinorhizobium meliloti / Q000378:metabolism*; D019802:Succinic Acid / Q000378:metabolism
D001426:Bacterial Proteins; D002352:Carrier Proteins; D029382:Dicarboxylic Acid Transporters; D003998:Dicarboxylic Acids; C073786:dctA protein, Sinorhizobium meliloti; D009963:Orotic Acid; C001242:5-fluoroorotic acid; D019802:Succinic Acid