Biochemical Characterization of the C4-dicarboxylate Transporter DctA from Bacillus Subtilis

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2010 Apr 6

PMID 20363944

Citations 22

Authors

Maarten Groeneveld

Ruud G J Detert Oude Weme

Ria H Duurkens

Dirk Jan Slotboom

Affiliations

Soon will be listed here.

Abstract

Bacterial secondary transporters of the DctA family mediate ion-coupled uptake of C(4)-dicarboxylates. Here, we have expressed the DctA homologue from Bacillus subtilis in the Gram-positive bacterium Lactococcus lactis. Transport of dicarboxylates in vitro in isolated membrane vesicles was assayed. We determined the substrate specificity, the type of cotransported ions, the electrogenic nature of transport, and the pH and temperature dependence patterns. DctA was found to catalyze proton-coupled symport of the four C(4)-dicarboxylates from the Krebs cycle (succinate, fumurate, malate, and oxaloacetate) but not of other mono- and dicarboxylates. Because (i) succinate-proton symport was electrogenic (stimulated by an internal negative membrane potential) and (ii) the divalent anionic form of succinate was recognized by DctA, at least three protons must be cotransported with succinate. The results were interpreted in the light of the crystal structure of the homologous aspartate transporter Glt(Ph) from Pyrococcus horikoshii.

Citing Articles

Deorphanizing solute carriers in for secondary uptake of xenobiotic compounds.

Moller-Hansen I, Saez-Saez J, van der Hoek S, Dyekjaer J, Christensen H, Muelas M Front Microbiol. 2024; 15:1376653.

PMID: 38680917 PMC: 11045925. DOI: 10.3389/fmicb.2024.1376653.

Evolutionary analysis reveals the origin of sodium coupling in glutamate transporters.

Reddy K, Rasool B, Akher F, Kutlesic N, Pant S, Boudker O bioRxiv. 2023; .

PMID: 38106174 PMC: 10723334. DOI: 10.1101/2023.12.03.569786.

K-12 Transcriptomics for Assessing the Mechanism of Action of High-Power Ultrasound.

Spiteri D, Griffin S, Karatzas K, Scerri C, Valdramidis V Microorganisms. 2023; 11(11).

PMID: 38004779 PMC: 10673019. DOI: 10.3390/microorganisms11112768.

A comparison of transporter gene expression in three species of Peronospora plant pathogens during host infection.

Johnson E, Lyon R, Zaitlin D, Khan A, Jairajpuri M PLoS One. 2023; 18(6):e0285685.

PMID: 37262030 PMC: 10234565. DOI: 10.1371/journal.pone.0285685.

Unchanged nitrate and nitrite isotope fractionation during heterotrophic and Fe(II)-mixotrophic denitrification suggest a non-enzymatic link between denitrification and Fe(II) oxidation.

Visser A, Wankel S, Frey C, Kappler A, Lehmann M Front Microbiol. 2022; 13:927475.

PMID: 36118224 PMC: 9478938. DOI: 10.3389/fmicb.2022.927475.

References

Grewer C, Balani P, Weidenfeller C, Bartusel T, Tao Z, Rauen T . Individual subunits of the glutamate transporter EAAC1 homotrimer function independently of each other. Biochemistry. 2005; 44(35):11913-23. PMC: 2459315. DOI: 10.1021/bi050987n. View

Kunji E, Slotboom D, Poolman B . Lactococcus lactis as host for overproduction of functional membrane proteins. Biochim Biophys Acta. 2003; 1610(1):97-108. DOI: 10.1016/s0005-2736(02)00712-5. View

Yurgel S, Mortimer M, Rogers K, Kahn M . New substrates for the dicarboxylate transport system of Sinorhizobium meliloti. J Bacteriol. 2000; 182(15):4216-21. PMC: 101915. DOI: 10.1128/JB.182.15.4216-4221.2000. View

Yarosh O, Charles T, Finan T . Analysis of C4-dicarboxylate transport genes in Rhizobium meliloti. Mol Microbiol. 1989; 3(6):813-23. DOI: 10.1111/j.1365-2958.1989.tb00230.x. View

Reyes N, Ginter C, Boudker O . Transport mechanism of a bacterial homologue of glutamate transporters. Nature. 2009; 462(7275):880-5. PMC: 2934767. DOI: 10.1038/nature08616. View

Koch H, Brown R, Larsson H . The glutamate-activated anion conductance in excitatory amino acid transporters is gated independently by the individual subunits. J Neurosci. 2007; 27(11):2943-7. PMC: 2435202. DOI: 10.1523/JNEUROSCI.0118-07.2007. View

Janausch I, Zientz E, Tran Q, Kroger A, Unden G . C4-dicarboxylate carriers and sensors in bacteria. Biochim Biophys Acta. 2002; 1553(1-2):39-56. DOI: 10.1016/s0005-2728(01)00233-x. View

Kuipers O, Beerthuyzen M, Siezen R, de Vos W . Characterization of the nisin gene cluster nisABTCIPR of Lactococcus lactis. Requirement of expression of the nisA and nisI genes for development of immunity. Eur J Biochem. 1993; 216(1):281-91. DOI: 10.1111/j.1432-1033.1993.tb18143.x. View

Endou H, Kanai Y . Cloning and functional characterization of a system ASC-like Na+-dependent neutral amino acid transporter. J Biol Chem. 1996; 271(25):14883-90. DOI: 10.1074/jbc.271.25.14883. View

10.

Levy L, Warr O, Attwell D . Stoichiometry of the glial glutamate transporter GLT-1 expressed inducibly in a Chinese hamster ovary cell line selected for low endogenous Na+-dependent glutamate uptake. J Neurosci. 1998; 18(23):9620-8. PMC: 6793325. View

11.

Prakash S, Cooper G, Singhi S, Saier Jr M . The ion transporter superfamily. Biochim Biophys Acta. 2003; 1618(1):79-92. DOI: 10.1016/j.bbamem.2003.10.010. View

12.

Tolner B, Poolman B, Konings W . Characterization of the proton/glutamate symport protein of Bacillus subtilis and its functional expression in Escherichia coli. J Bacteriol. 1995; 177(10):2863-9. PMC: 176960. DOI: 10.1128/jb.177.10.2863-2869.1995. View

13.

Poolman B, Smid E, Konings W . Kinetic properties of a phosphate-bond-driven glutamate-glutamine transport system in Streptococcus lactis and Streptococcus cremoris. J Bacteriol. 1987; 169(6):2755-61. PMC: 212181. DOI: 10.1128/jb.169.6.2755-2761.1987. View

14.

Ronson C, Astwood P, Downie J . Molecular cloning and genetic organization of C4-dicarboxylate transport genes from Rhizobium leguminosarum. J Bacteriol. 1984; 160(3):903-9. PMC: 215795. DOI: 10.1128/jb.160.3.903-909.1984. View

15.

Leary G, Stone E, Holley D, Kavanaugh M . The glutamate and chloride permeation pathways are colocalized in individual neuronal glutamate transporter subunits. J Neurosci. 2007; 27(11):2938-42. PMC: 6672579. DOI: 10.1523/JNEUROSCI.4851-06.2007. View

16.

Koch H, Larsson H . Small-scale molecular motions accomplish glutamate uptake in human glutamate transporters. J Neurosci. 2005; 25(7):1730-6. PMC: 6725926. DOI: 10.1523/JNEUROSCI.4138-04.2005. View

17.

Groeneveld M, Slotboom D . Rigidity of the subunit interfaces of the trimeric glutamate transporter GltT during translocation. J Mol Biol. 2007; 372(3):565-70. DOI: 10.1016/j.jmb.2007.06.067. View

18.

van Slooten J, Bhuvanasvari T, Bardin S, Stanley J . Two C4-dicarboxylate transport systems in Rhizobium sp. NGR234: rhizobial dicarboxylate transport is essential for nitrogen fixation in tropical legume symbioses. Mol Plant Microbe Interact. 1992; 5(2):179-86. DOI: 10.1094/mpmi-5-179. View

19.

Eudes A, Erkens G, Slotboom D, Rodionov D, Naponelli V, Hanson A . Identification of genes encoding the folate- and thiamine-binding membrane proteins in Firmicutes. J Bacteriol. 2008; 190(22):7591-4. PMC: 2576666. DOI: 10.1128/JB.01070-08. View

20.

Davies S, Golby P, Omrani D, Broad S, Harrington V, Guest J . Inactivation and regulation of the aerobic C(4)-dicarboxylate transport (dctA) gene of Escherichia coli. J Bacteriol. 1999; 181(18):5624-35. PMC: 94081. DOI: 10.1128/JB.181.18.5624-5635.1999. View