Abstract
Neisseria meningitidis is a pathogenic bacterium responsible for meningitis. The mechanisms underlying the control of Na+ transmembrane movement, presumably important to pathogenicity, have been barely addressed. To elucidate the function of the components of the Na+ transport system in N. meningitidis, an open reading frame from the genome of this bacterium displaying similarity with the NhaE type of Na+/H+ antiporters was expressed in Escherichia coli and characterized for sodium transport ability. The N. meningitidis antiporter (NmNhaE) was able to complement an E. coli strain devoid of Na+/H+ antiporters (KNabc) respecting the ability to grow in the presence of NaCl and LiCl. Ion transport assays in everted vesicles prepared from KNabc expressing NmNhaE from a plasmid confirmed its ability to translocate Na+ and Li+. Here is presented the characterization of the first NhaE from a pathogen, an important contribution to the comprehension of sodium ion metabolism in this kind of microorganisms.
This is a preview of subscription content, log in via an institution to check access.
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Ausubel FM et al (1995) Current protocols in molecular biology. Greene Publishing Associates/Wiley Interscience, New York
Bernsel A, Viklund H, Hennerdal A, Elofsson A (2009) TOPCONS: consensus prediction of membrane protein topology. Nucleic Acids Res 37:W465–W468
Dzioba J, Ostroumov E, Winogrodzki A, Dibrov P (2002) Cloning, functional expression in Escherichia coli and primary characterization of a new Na+/H+ antiporter, NhaD, of Vibrio cholerae. Mol Cell Biochem 229:119–124
Dzioba-Winogrodzki J, Winogrodzki O, Krulwich TA, Boin MA, Häse CC, Dibrov P (2009) The Vibrio cholerae Mrp system: cation/proton antiport properties and enhancement of bile salt resistance in a heterologous host. J Mol Microbiol Biotechnol 16:176–186
Goldberg EB et al (1987) Characterization of a Na+/H+ antiporter gene of Escherichia coli. Proc Natl Acad Sci USA 84:2615–2619
Harrison LH, Trotter CL, Ramsay ME (2009) Global epidemiology of meningococcal disease. Vaccine 27(Suppl 2):B51–B63
Häse CC, Fedorova ND, Galperin MY, Dibrov PA (2001) Sodium ion cycle in bacterial pathogens: evidence from cross-genome comparisons. Microbiol Mol Biol Rev 65:353–370
Herz K, Vimont S, Padan E, Berche P (2003) Roles of NhaA, NhaB, and NhaD Na+/H+ antiporters in survival of Vibrio cholerae in a saline environment. J Bacteriol 185:1236–1244
Hirokawa T, Boon-Chieng S, Mitaku S (1998) SOSUI: classification and secondary structure prediction system for membrane proteins. Bioinformatics 14:378–379
Hunte C, Screpanti E, Venturi M, Rimon A, Padan E, Michel H (2005) Structure of a Na+/H+ antiporter and insights into mechanism of action and regulation by pH. Nature 435:1197–1202
Inoue H, Sakurai T, Ujike S, Tsuchiya T, Murakami H, Kanazawa H (1999) Expression of functional Na+/H+ antiporters of Helicobacter pylori in antiporter-deficient Escherichia coli mutants. FEBS Lett 443:11–16
Ito M, Guffanti AA, Zemsky J, Ivey DM, Krulwich TA (1997) Role of the nhaC-encoded Na+/H+ antiporter of alkaliphilic Bacillus firmus OF4. J Bacteriol 179:3851–3857
Kanazawa H, Miki T, Tamura F, Yura T, Futai M (1979) Specialized transducing phage lambda carrying the genes for coupling factor of oxidative phosphorylation of Escherichia coli: increased synthesis of coupling factor on induction of prophage lambda asn. Proc Natl Acad Sci USA 76:1126–1130
Lehmann WD, Bahr U, Schulten HR (1978) Determination of lithium in microlitre amounts of human body fluids at therapeutic and normal levels by stable isotope dilution and field desorption mass spectrometry. Biomed Mass Spectrom 5:536–539
Lineweaver H, Burk D (1934) The determination of enzyme dissociation constants. J Am Chem Soc 56:658–666
McGuffin LJ, Bryson K, Jones DT (2000) The PSIPRED protein structure prediction server. Bioinformatics 16:404–405
Melo AMP et al (2005) A nhaD Na+/H+ antiporter and a pcd homologues are among the Rhodothermus marinus complex I genes. Biochim Biophys Acta 1709:95–103
Melo AMP, Felix NA, Carita JN, Saraiva LM, Teixeira M (2006) The Na+/H+ antiporter of the thermohalophilic bacterium Rhodothermus marinus. Biochem Biophys Res Commun 348:1011–1017
Nozaki K, Inaba K, Kuroda T, Tsuda M, Tsuchiya T (1996) Cloning and sequencing of the gene for Na+/H+ antiporter of Vibrio parahaemolyticus. Biochem Biophys Res Commun 222:774–779
Padan E, Venturi M, Gerchman Y, Dover N (2001) Na(+)/H(+) antiporters. Biochim Biophys Acta 1505:144–157
Pinner E, Padan E, Schuldiner S (1992) Cloning, sequencing, and expression of the nhaB gene, encoding a Na+/H+ antiporter in Escherichia coli. J Biol Chem 267:11064–11068
Pinner E, Kotler Y, Padan E, Schuldiner S (1993) Physiological role of NhaB, a specific Na+/H+ antiporter in Escherichia coli. J Biol Chem 268:1729–1734
Pinner E, Padan E, Schuldiner S (1994) Kinetic properties of NhaB, a Na+/H+ antiporter from Escherichia coli. J Biol Chem 269:26274–26279
Rosen BP (1986) Ion extrusion systems in Escherichia coli. Methods Enzymol 125:328–336
Sutherland TC, Quattroni P, Exley RM, Tang CM (2010) Transcellular passage of Neisseria meningitidis across a polarized respiratory epithelium. Infect Immun 78:3832–3847
Swartz TH, Ikewada S, Ishikawa O, Ito M, Krulwich TA (2005) The Mrp system: a giant among monovalent cation/proton antiporters? Extremophiles 9:345–354
Taglicht D, Padan E, Schuldiner S (1991) Overproduction and purification of a functional Na+/H+ antiporter coded by nhaA (ant) from Escherichia coli. J Biol Chem 266:11289–11294
Tettelin H et al (2000) Complete genome sequence of Neisseria meningitidis serogroup B strain MC58. Science 287:1809–1815
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Vorburger T, Stein A, Ziegler U, Kaim G, Steuber J (2009) Functional role of a conserved aspartic acid residue in the motor of the Na(+)-driven flagellum from Vibrio cholerae. Biochim Biophys Acta 1787:1198–1204
White AG, Entmacher PS, Rubin G, Leiter L (1955) Physiological and pharmacological regulation of human salivary electrolyte concentrations; with a discussion of electrolyte concentrations of some other exocrine secretions. J Clin Invest 34:246–255
Acknowledgments
We are grateful to Drs Christoph Tang and Hiroshi Kanazawa for the generous gift of N. meningitidis genomic DNA and the E. coli strain KNabc, respectively. Filipe Santos and Andreia Rodrigues are acknowledged for excellent technical assistance. AMPM was awarded a Deutcher Akademischer Austausch Dienst research visit grant and PMFS received a grant from Fundação para a Ciência e a Tecnologia (SFRH/BD/46553/2008).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by Krämer Reinhard.
Electronic supplementary material
Below is the link to the electronic supplementary material.
203_2012_856_MOESM1_ESM.docx
Alignment of representative amino acid sequences from the five families of Nha Na+/H+ antiporters obtained using ClustalW. Aa, Alkalimonas amylolytica, NhaD (AAX63482.1); Bf, Bacillus firmus, NhaC (AAC45432.1); Bsp., Bacillus sp., NhaC (ABH03020.1); Dv, Desulfovibrio vulgaris, NhaE (YP_009252.1); Ec, Escherichia coli, NhaA (NP_414560.1), NhaB (AAA24218.1|); Ef, Enterococcus faecalis, NhaC (ZP_05501989.1); He, Halomonas elongata, NhaD (YP_003897392.1); Hi, Haemophilus influenzae, NhaB (ZP_05848338.1); Hp, Helicobacter pylori, NhaA (O26076.1); Kp, Klebsiella pneumoniae, NhaB (YP_002237820.1); Mc, Methylococcus capsulatus, NhaD (YP_114201.1); Mt, Moorella thermoacetica, NhaE (YP_429442.1); Nm, Neisseria meningitidis, NhaC (NP_273581.1), NhaE (NP_273614.1); Psp., Pseudomonas sp., NhaA (AAV49149.1); Rm, Rhodothermus marinus, NhaE (AAY42129.1); Vc, Vibrio cholerae, NhaA (O85187.1), NhaB (Q9KQU7.1), NhaD (AAG48354.2). (DOCX 40 kb)
203_2012_856_MOESM2_ESM.docx
Primary sequence comparison of the 50 closest hits from a GenBank search using NmNhaE as query. Amoebophilus asiaticus, Amoebophilus asiaticus (YP_001958297.1); A sp1., Algoriphagus sp. (ZP_01718340.1); A sp2., Anaeromyxobacter sp. (YP_001380997.1); A vinelandii, Azotobacter vinelandii (YP_002798245.1); B Ellin514, bacterium Ellin 514 (ZP_03629791.1);B japonicum, Bradyrhizobium japonicum (NP_773762.1); B phymatum, Burkholderia phymatum (YP_001857045.1); B sp., Burkholderia sp.(YP_368456.1); C eutactus, Coprococcus eutactus (ZP_02207858.1); C flavus, Chthoniobacter flavus(ZP_03133069.1); C testosteroni, Comamonas testosteroni (ZP_03544348.1); C limicola, Chlorobium limicola (YP_001942426.1); C phaeobacte, Chlorobium phaeobacteroides (YP_001960386.1); C tepidum, Chlorobium tepidum (NP_661198.1); D acidovorans, Delftia acidovorans (YP_001565753.1); D baculatum, Desulfomicrobium baculatum (YP_003159209.1); D hafniense, Desulfitobacterium hafniense (YP_520285.1); D sp., Diaphorobacter sp.(YP_002552833.1); D vulgaris, Desulfovibrio vulgaris (YP_009252.1); E corrodens, Eikenella corrodens (ZP_03713713.1); E hallii, Eubacterium hallii (ZP_03715974.1); H arsenic, Herminiimonas arsenicoxydans (YP_001099123.1); K oralis, Kingella oralis (ZP_04602428.1); L cholodnii, Leptothrix cholodnii(YP_001791022.1); L hongkongens, Laribacter hongkongensis (YP_002795798.1); L intracell, Lawsonia intracellularis (YP_594890.1); L nitroferrum, Lutiella nitroferrum (ZP_03697702.1 and ZP_03699435.1); J sp., Janthinobacterium sp. (YP_001352410.1); Nm, Neisseria meningitidis (NP_273614.1); M loti, Mesorhizobium loti (NP_105654.1); M petroleiph, Methylibium petroleiphilum (YP_001020707.1); M silvestris, Methylocella silvestris (YP_002362098.1); M thermoacetica, Moorella thermoacetica (YP_429442.1); O carboxido, Oligotropha carboxidovorans (YP_002290536.1); P aestuarii, Prosthecochloris aestuarii (YP_002016448.1); P phaeoc, Pelodictyon phaeoclathratiforme (YP_002017089.1); P naphthalen, Polaromonas naphthalenivorans (YP_981856.1); T intermédia, Thiomonas intermédia (ZP_05500427.1); R centenum, Rhodospirillum centenum (YP_002297310.1); R eutropha, Ralstonia eutropha (YP_727038.1); R intestinalis, Roseburia intestinalis (ZP_04745272.1); R marinus, Rhodothermus marinus, (AAY42129.1); R palustris, Rhodopseudomonas palustris (ZP_06360325.1); R sp., Ruminococcus sp. (ZP_04857516.1); S fumaroxidan, Syntrophobacter fumaroxidans (YP_845285.1); V eiseniae, Verminephrobacter eiseniae (YP_996836.1); V paradoxus, Variovorax paradoxus (YP_002943689.1); V spinosum, Verrucomicrobium spinosum (ZP_02926838.1); X autotrophi, Xanthobacter autotrophicus (YP_001416350.1). (DOCX 97 kb)
Rights and permissions
About this article
Cite this article
Sousa, P.M.F., Videira, M.A.M., Vorburger, T. et al. The novel NhaE-type Na+/H+ antiporter of the pathogenic bacterium Neisseria meningitidis . Arch Microbiol 195, 211–217 (2013). https://doi.org/10.1007/s00203-012-0856-4
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00203-012-0856-4