Secondary transporters for citrate and the Mg(2+)-citrate complex in Bacillus subtilis are homologous proteins.
Citrate uptake in Bacillus subtilis is mediated by a secondary transporter that transports the complex of citrate and divalent metal ions. The gene coding for the transporter termed CitM was cloned, sequenced, and functionally expressed in Escherichia coli. Translation of the base sequence to the primary sequence revealed a transporter that is not homologous to any known secondary transporter. However, CitM shares 60% sequence identity with the gene product of open reading frame N15CR that is on the genome of B. subtilis and for which no function is known. The hydropathy profiles of the primary sequences of CitM and the unknown gene product are very similar, and secondary structure prediction algorithms predict 12 transmembrane-spanning segments for both proteins. Open reading frame N15CR was cloned and expressed in E. coli and was shown to be a citrate transporter as well. The transporter is termed CitH. A remarkable difference between the two transporters is that citrate uptake by CitM is stimulated by the presence of Mg2+ ions, while citrate uptake by CitH is inhibited by Mg2+. It is concluded that the substrate of CitM is the Mg(2+)-citrate complex and that CitH transports the free citrate anion. Uptake experiments in right-side-out membrane vesicles derived from E. coli cells expressing either CitM or CitH showed that both transporters catalyze electrogenic proton/substrate symport.
Department of Microbiology, Groningen Biotechnology and Biomolecular Sciences Institute, University of Groningen, The Netherlands.
A
A
Boorsma
M E
ME
van der Rest
J S
JS
Lolkema
W N
WN
Konings
D001412:Bacillus subtilis / Q000378:metabolism*; D001426:Bacterial Proteins; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics*; D002951:Citrates / Q000378:metabolism*; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial; D007477:Ions; D008274:Magnesium / Q000378:metabolism*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D027981:Symporters
D001426:Bacterial Proteins; D002352:Carrier Proteins; C102671:CitM protein, Bacillus subtilis; D002951:Citrates; D017931:DNA Primers; D004269:DNA, Bacterial; D007477:Ions; D027981:Symporters; D008274:Magnesium