NHAA_ECOLI
ID NHAA_ECOLI Reviewed; 388 AA.
AC P13738;
DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1996, sequence version 3.
DT 03-AUG-2022, entry version 180.
DE RecName: Full=Na(+)/H(+) antiporter NhaA {ECO:0000303|PubMed:2839489};
DE AltName: Full=Na(+)/H(+) exchanger {ECO:0000303|PubMed:19396973};
DE AltName: Full=Na(+)/Li(+)/H(+) antiporter {ECO:0000303|PubMed:8019504};
DE AltName: Full=Sodium/proton antiporter NhaA {ECO:0000305};
GN Name=nhaA {ECO:0000303|PubMed:1645730};
GN Synonyms=ant {ECO:0000303|PubMed:2839489}; OrderedLocusNames=b0019, JW0018;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RX PubMed=2839489; DOI=10.1016/s0021-9258(19)81531-4;
RA Karpel R., Olami Y., Taglicht D., Schuldiner S., Padan E.;
RT "Sequencing of the gene ant which affects the Na+/H+ antiporter activity in
RT Escherichia coli.";
RL J. Biol. Chem. 263:10408-10414(1988).
RN [2]
RP SEQUENCE REVISION.
RA Padan E.;
RL Submitted (FEB-2000) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=1630901; DOI=10.1093/nar/20.13.3305;
RA Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K.,
RA Mizobuchi K., Nakata A.;
RT "Systematic sequencing of the Escherichia coli genome: analysis of the 0-
RT 2.4 min region.";
RL Nucleic Acids Res. 20:3305-3308(1992).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-28, AND INDUCTION.
RC STRAIN=K12;
RX PubMed=1657980; DOI=10.1016/s0021-9258(18)54700-1;
RA Karpel R., Alon T., Glaser G., Schuldiner S., Padan E.;
RT "Expression of a sodium proton antiporter (NhaA) in Escherichia coli is
RT induced by Na+ and Li+ ions.";
RL J. Biol. Chem. 266:21753-21759(1991).
RN [7]
RP PROTEIN SEQUENCE OF 1-10, FUNCTION, ACTIVITY REGULATION, AND SUBCELLULAR
RP LOCATION.
RC STRAIN=K12;
RX PubMed=1645730; DOI=10.1016/s0021-9258(18)99161-1;
RA Taglicht D., Padan E., Schuldiner S.;
RT "Overproduction and purification of a functional Na+/H+ antiporter coded by
RT nhaA (ant) from Escherichia coli.";
RL J. Biol. Chem. 266:11289-11294(1991).
RN [8]
RP FUNCTION, AND CATALYTIC ACTIVITY.
RX PubMed=8383669; DOI=10.1016/s0021-9258(18)53333-0;
RA Taglicht D., Padan E., Schuldiner S.;
RT "Proton-sodium stoichiometry of NhaA, an electrogenic antiporter from
RT Escherichia coli.";
RL J. Biol. Chem. 268:5382-5387(1993).
RN [9]
RP MUTAGENESIS OF HISTIDINE RESIDUES.
RX PubMed=8381959; DOI=10.1073/pnas.90.4.1212;
RA Gerchman Y., Olami Y., Rimon A., Taglicht D., Schuldiner S., Padan E.;
RT "Histidine-226 is part of the pH sensor of NhaA, a Na+/H+ antiporter in
RT Escherichia coli.";
RL Proc. Natl. Acad. Sci. U.S.A. 90:1212-1216(1993).
RN [10]
RP FUNCTION IN SODIUM AND LITHIUM TRANSPORT, BIOPHYSICOCHEMICAL PROPERTIES,
RP AND DISRUPTION PHENOTYPE.
RC STRAIN=K12;
RX PubMed=8019504; DOI=10.1248/bpb.17.395;
RA Inaba K., Kuroda T., Shimamoto T., Kayahara T., Tsuda M., Tsuchiya T.;
RT "Lithium toxicity and Na+(Li+)/H+ antiporter in Escherichia coli.";
RL Biol. Pharm. Bull. 17:395-398(1994).
RN [11]
RP FUNCTION, AND MUTAGENESIS OF ASP-65; ASP-133; ASP-163; ASP-164 AND ASP-282.
RX PubMed=7737413; DOI=10.1016/0014-5793(95)00331-3;
RA Inoue H., Noumi T., Tsuchiya T., Kanazawa H.;
RT "Essential aspartic acid residues, Asp-133, Asp-163 and Asp-164, in the
RT transmembrane helices of a Na+/H+ antiporter (NhaA) from Escherichia
RT coli.";
RL FEBS Lett. 363:264-268(1995).
RN [12]
RP MUTAGENESIS OF HIS-225.
RX PubMed=7592922; DOI=10.1074/jbc.270.45.26813;
RA Rimon A., Gerchman Y., Olami Y., Schuldiner S., Padan E.;
RT "Replacements of histidine 226 of NhaA-Na+/H+ antiporter of Escherichia
RT coli. Cysteine (H226C) or serine (H226S) retain both normal activity and pH
RT sensitivity, aspartate (H226D) shifts the pH profile toward basic pH, and
RT alanine (H226A) inactivates the carrier at all pH values.";
RL J. Biol. Chem. 270:26813-26817(1995).
RN [13]
RP ACTIVITY REGULATION, AND MUTAGENESIS OF GLU-241; 242-LYS--HIS-253; LYS-249
RP AND VAL-254.
RX PubMed=10455127; DOI=10.1074/jbc.274.35.24617;
RA Gerchman Y., Rimon A., Padan E.;
RT "A pH-dependent conformational change of NhaA Na(+)/H(+) antiporter of
RT Escherichia coli involves loop VIII-IX, plays a role in the pH response of
RT the protein, and is maintained by the pure protein in dodecyl maltoside.";
RL J. Biol. Chem. 274:24617-24624(1999).
RN [14]
RP SUBUNIT, AND MUTAGENESIS OF ASP-163; ASP-164; HIS-225 AND GLY-338.
RX PubMed=11258962; DOI=10.1021/bi002669o;
RA Gerchman Y., Rimon A., Venturi M., Padan E.;
RT "Oligomerization of NhaA, the Na+/H+ antiporter of Escherichia coli in the
RT membrane and its functional and structural consequences.";
RL Biochemistry 40:3403-3412(2001).
RN [15]
RP ACTIVITY REGULATION, AND MUTAGENESIS OF GLU-252.
RX PubMed=14604993; DOI=10.1074/jbc.m309021200;
RA Tzubery T., Rimon A., Padan E.;
RT "Mutation E252C increases drastically the Km value for Na+ and causes an
RT alkaline shift of the pH dependence of NhaA Na+/H+ antiporter of
RT Escherichia coli.";
RL J. Biol. Chem. 279:3265-3272(2004).
RN [16]
RP SUBCELLULAR LOCATION, AND TOPOLOGY [LARGE SCALE ANALYSIS].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=15919996; DOI=10.1126/science.1109730;
RA Daley D.O., Rapp M., Granseth E., Melen K., Drew D., von Heijne G.;
RT "Global topology analysis of the Escherichia coli inner membrane
RT proteome.";
RL Science 308:1321-1323(2005).
RN [17]
RP SUBUNIT, AND MUTAGENESIS OF 45-PRO--ASN-58.
RX PubMed=17635927; DOI=10.1074/jbc.m704469200;
RA Rimon A., Tzubery T., Padan E.;
RT "Monomers of the NhaA Na+/H+ antiporter of Escherichia coli are fully
RT functional yet dimers are beneficial under extreme stress conditions at
RT alkaline pH in the presence of Na+ or Li+.";
RL J. Biol. Chem. 282:26810-26821(2007).
RN [18]
RP MUTAGENESIS OF ASP-163 AND ASP-164.
RX PubMed=19274728; DOI=10.1002/prot.22368;
RA Olkhova E., Kozachkov L., Padan E., Michel H.;
RT "Combined computational and biochemical study reveals the importance of
RT electrostatic interactions between the 'pH sensor' and the cation binding
RT site of the sodium/proton antiporter NhaA of Escherichia coli.";
RL Proteins 76:548-559(2009).
RN [19]
RP FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, AND MUTAGENESIS OF THR-132;
RP ASP-133; ASP-163; ASP-164 AND LYS-300.
RX PubMed=22915592; DOI=10.1074/jbc.m112.391128;
RA Maes M., Rimon A., Kozachkov-Magrisso L., Friedler A., Padan E.;
RT "Revealing the ligand binding site of NhaA Na+/H+ antiporter and its pH
RT dependence.";
RL J. Biol. Chem. 287:38150-38157(2012).
RN [20]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, SUBUNIT, AND MUTAGENESIS
RP OF 45-PRO--ASN-58; ALA-167; HIS-225 AND VAL-254.
RX PubMed=23836890; DOI=10.1074/jbc.m113.484071;
RA Mager T., Braner M., Kubsch B., Hatahet L., Alkoby D., Rimon A., Padan E.,
RA Fendler K.;
RT "Differential effects of mutations on the transport properties of the
RT Na+/H+ antiporter NhaA from Escherichia coli.";
RL J. Biol. Chem. 288:24666-24675(2013).
RN [21]
RP DOMAIN.
RX PubMed=22694117; DOI=10.3109/09687688.2012.693209;
RA Kozachkov L., Padan E.;
RT "Conformational changes in NhaA Na+/H+ antiporter.";
RL Mol. Membr. Biol. 30:90-100(2013).
RN [22]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF ASP-133 AND ALA-167.
RX PubMed=27021484; DOI=10.1038/srep23339;
RA Dwivedi M., Sukenik S., Friedler A., Padan E.;
RT "The Ec-NhaA antiporter switches from antagonistic to synergistic antiport
RT upon a single point mutation.";
RL Sci. Rep. 6:23339-23339(2016).
RN [23]
RP DOMAIN, AND PROPOSED MECHANISM.
RX PubMed=27708266; DOI=10.1038/ncomms12940;
RA Huang Y., Chen W., Dotson D.L., Beckstein O., Shen J.;
RT "Mechanism of pH-dependent activation of the sodium-proton antiporter
RT NhaA.";
RL Nat. Commun. 7:12940-12940(2016).
RN [24]
RP DOMAIN, AND MUTAGENESIS OF LYS-300.
RX PubMed=28330875; DOI=10.1074/jbc.m117.778175;
RA Calinescu O., Dwivedi M., Patino-Ruiz M., Padan E., Fendler K.;
RT "Lysine 300 is essential for stability but not for electrogenic transport
RT of the Escherichia coli NhaA Na+/H+ antiporter.";
RL J. Biol. Chem. 292:7932-7941(2017).
RN [25]
RP MUTAGENESIS OF ASP-133.
RX PubMed=29410365; DOI=10.1016/j.jmb.2018.01.014;
RA Rimon A., Dwivedi M., Friedler A., Padan E.;
RT "Asp133 residue in NhaA Na+/H+ antiporter is required for stability cation
RT binding and transport.";
RL J. Mol. Biol. 430:867-880(2018).
RN [26]
RP MUTAGENESIS OF ASP-163 AND LYS-300, AND ALTERNATIVE TRANSPORT MECHANISMS.
RX PubMed=30409911; DOI=10.1074/jbc.ra118.004903;
RA Patino-Ruiz M., Dwivedi M., Calinescu O., Karabel M., Padan E., Fendler K.;
RT "Replacement of Lys-300 with a glutamine in the NhaA Na+/H+ antiporter of
RT Escherichia coli yields a functional electrogenic transporter.";
RL J. Biol. Chem. 294:246-256(2019).
RN [27]
RP REVIEW, AND COMPARATIVE ANALYSIS OF VARIOUS MUTATIONS.
RX PubMed=33129932; DOI=10.1016/j.biochi.2020.10.017;
RA Dwivedi M.;
RT "Site-directed mutations reflecting functional and structural properties of
RT Ec-NhaA.";
RL Biochimie 180:79-89(2021).
RN [28] {ECO:0007744|PDB:1ZCD}
RP X-RAY CRYSTALLOGRAPHY (3.45 ANGSTROMS), SUBUNIT, SUBCELLULAR LOCATION,
RP TOPOLOGY, AND DOMAIN.
RX PubMed=15988517; DOI=10.1038/nature03692;
RA Hunte C., Screpanti E., Venturi M., Rimon A., Padan E., Michel H.;
RT "Structure of a Na+/H+ antiporter and insights into mechanism of action and
RT regulation by pH.";
RL Nature 435:1197-1202(2005).
RN [29] {ECO:0007744|PDB:3FI1}
RP STRUCTURE BY ELECTRON MICROSCOPY (7.00 ANGSTROMS) OF 9-384, SUBUNIT,
RP SUBCELLULAR LOCATION, TOPOLOGY, AND DOMAIN.
RX PubMed=19396973; DOI=10.1016/j.jmb.2009.03.010;
RA Appel M., Hizlan D., Vinothkumar K.R., Ziegler C., Kuehlbrandt W.;
RT "Conformations of NhaA, the Na+/H+ exchanger from Escherichia coli, in the
RT pH-activated and ion-translocating states.";
RL J. Mol. Biol. 388:659-672(2009).
RN [30] {ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5}
RP X-RAY CRYSTALLOGRAPHY (3.50 ANGSTROMS) OF WILD-TYPE AND MUTANT
RP THR-109/GLY-277/MET-296, SUBUNIT, SUBCELLULAR LOCATION, TOPOLOGY, DOMAIN,
RP AND PROPOSED MECHANISM.
RX PubMed=25422503; DOI=10.1085/jgp.201411219;
RA Lee C., Yashiro S., Dotson D.L., Uzdavinys P., Iwata S., Sansom M.S.,
RA von Ballmoos C., Beckstein O., Drew D., Cameron A.D.;
RT "Crystal structure of the sodium-proton antiporter NhaA dimer and new
RT mechanistic insights.";
RL J. Gen. Physiol. 144:529-544(2014).
CC -!- FUNCTION: Na(+)/H(+) antiporter that extrudes sodium in exchange for
CC external protons (PubMed:2839489, PubMed:1645730, PubMed:8383669,
CC PubMed:8019504, PubMed:7737413, PubMed:23836890). Plays an important
CC role in the regulation of intracellular pH, cellular Na(+) content and
CC cell volume (PubMed:33129932). Catalyzes the exchange of 2 H(+) per
CC Na(+) (PubMed:8383669, PubMed:23836890). This stoichiometry applies at
CC both neutral and alkaline pH values (PubMed:8383669). In addition, can
CC also transport lithium and is involved in lithium detoxification
CC (PubMed:8019504, PubMed:7737413, PubMed:22915592, PubMed:27021484).
CC Binding of the Li(+) and H(+) ligands to NhaA is coupled and
CC antagonistic (PubMed:27021484). {ECO:0000269|PubMed:1645730,
CC ECO:0000269|PubMed:22915592, ECO:0000269|PubMed:23836890,
CC ECO:0000269|PubMed:27021484, ECO:0000269|PubMed:2839489,
CC ECO:0000269|PubMed:7737413, ECO:0000269|PubMed:8019504,
CC ECO:0000269|PubMed:8383669, ECO:0000303|PubMed:33129932}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 H(+)(out) + Na(+)(in) = 2 H(+)(in) + Na(+)(out);
CC Xref=Rhea:RHEA:29251, ChEBI:CHEBI:15378, ChEBI:CHEBI:29101;
CC Evidence={ECO:0000269|PubMed:23836890, ECO:0000269|PubMed:8383669};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:29252;
CC Evidence={ECO:0000269|PubMed:23836890, ECO:0000269|PubMed:8383669};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 H(+)(out) + Li(+)(in) = 2 H(+)(in) + Li(+)(out);
CC Xref=Rhea:RHEA:70431, ChEBI:CHEBI:15378, ChEBI:CHEBI:49713;
CC Evidence={ECO:0000269|PubMed:27021484, ECO:0000305|PubMed:8019504};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70432;
CC Evidence={ECO:0000269|PubMed:27021484, ECO:0000305|PubMed:8019504};
CC -!- ACTIVITY REGULATION: Activity is regulated by pH (PubMed:1645730,
CC PubMed:23836890, PubMed:10455127, PubMed:14604993). Active at alkaline
CC pH (PubMed:1645730, PubMed:23836890, PubMed:10455127, PubMed:14604993).
CC Activity is strongly down-regulated below pH 6.5 and a dramatic
CC increase in activity is observed upon increase of the pH from 6.5 to
CC 8.5 (PubMed:1645730). {ECO:0000269|PubMed:10455127,
CC ECO:0000269|PubMed:14604993, ECO:0000269|PubMed:1645730,
CC ECO:0000269|PubMed:23836890}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.98 mM for Na(+) {ECO:0000269|PubMed:8019504};
CC KM=0.2 mM for Na(+) {ECO:0000269|PubMed:22915592};
CC KM=0.91 mM for Li(+) {ECO:0000269|PubMed:8019504};
CC KM=0.02 mM for Li(+) {ECO:0000269|PubMed:22915592};
CC -!- SUBUNIT: Monomer (PubMed:17635927). Homodimer (PubMed:11258962,
CC PubMed:17635927, PubMed:15988517, PubMed:19396973, PubMed:25422503).
CC Under routine stress conditions, the monomeric form is fully functional
CC (PubMed:17635927, PubMed:23836890). However, the dimeric form is much
CC more efficient in conferring growth resistance under extreme stress
CC conditions (PubMed:17635927). {ECO:0000269|PubMed:11258962,
CC ECO:0000269|PubMed:15988517, ECO:0000269|PubMed:17635927,
CC ECO:0000269|PubMed:19396973, ECO:0000269|PubMed:23836890,
CC ECO:0000269|PubMed:25422503}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane {ECO:0000269|PubMed:15919996,
CC ECO:0000269|PubMed:15988517, ECO:0000269|PubMed:1645730,
CC ECO:0000269|PubMed:19396973, ECO:0000269|PubMed:25422503}; Multi-pass
CC membrane protein {ECO:0000269|PubMed:15988517,
CC ECO:0000269|PubMed:19396973, ECO:0000269|PubMed:25422503}.
CC -!- INDUCTION: Expression is induced by Na(+) and Li(+). The Na(+)-
CC dependent enhancement of expression is augmented by alkalinization.
CC {ECO:0000269|PubMed:1657980}.
CC -!- DOMAIN: Displays a non-canonical transmembrane assembly, which is
CC termed the NhaA fold. A unique assembly of two pairs of short helices
CC connected by crossed, extended chains creates a balanced electrostatic
CC environment for the binding of the substrate ions (PubMed:15988517). Is
CC organized into two functional regions: the pH sensor region, a cluster
CC of amino-acyl side chains involved in pH regulation, and a catalytic
CC region containing the ion-binding site located approximately 9
CC Angstroms apart from the pH sensor region (PubMed:33129932).
CC {ECO:0000269|PubMed:15988517, ECO:0000303|PubMed:33129932}.
CC -!- DOMAIN: Shows two different conformational changes in response to pH
CC and substrate ions (PubMed:19396973, PubMed:22694117). The first change
CC is induced by a rise in pH from 6 to 7 and marks the transition to the
CC pH-activated state (PubMed:19396973). The second conformational change
CC is induced by the substrate ions Na(+) and Li(+) at pH above 7
CC (PubMed:19396973). This movement would cause a charge imbalance at the
CC ion-binding site that may trigger the release of the substrate ion and
CC open a periplasmic exit channel (PubMed:19396973). The transport
CC mechanism may involve Asp-163 switching between forming a salt bridge
CC with Lys-300 and interacting with the proton and/or sodium ion
CC (PubMed:25422503, PubMed:27708266, PubMed:28330875). The salt bridge
CC may stabilize the conformation (PubMed:28330875).
CC {ECO:0000269|PubMed:19396973, ECO:0000269|PubMed:22694117,
CC ECO:0000269|PubMed:25422503, ECO:0000269|PubMed:27708266,
CC ECO:0000269|PubMed:28330875}.
CC -!- DISRUPTION PHENOTYPE: Mutant lacking this gene can grow in the presence
CC of 0.1 M LiCl, but not in the presence of 0.6 M LiCl.
CC {ECO:0000269|PubMed:8019504}.
CC -!- SIMILARITY: Belongs to the NhaA Na(+)/H(+) (TC 2.A.33) antiporter
CC family. {ECO:0000255|HAMAP-Rule:MF_01844, ECO:0000305}.
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DR EMBL; J03879; AAA23448.2; -; Genomic_DNA.
DR EMBL; U00096; AAC73130.1; -; Genomic_DNA.
DR EMBL; AP009048; BAB96592.1; -; Genomic_DNA.
DR EMBL; S67239; AAB20348.1; -; Genomic_DNA.
DR PIR; C64722; C64722.
DR RefSeq; NP_414560.1; NC_000913.3.
DR RefSeq; WP_000681354.1; NZ_LN832404.1.
DR PDB; 1ZCD; X-ray; 3.45 A; A/B=1-388.
DR PDB; 3FI1; EM; 7.00 A; A=9-384.
DR PDB; 4ATV; X-ray; 3.50 A; A/B/C/D=1-388.
DR PDB; 4AU5; X-ray; 3.70 A; A/B/C/D=1-388.
DR PDBsum; 1ZCD; -.
DR PDBsum; 3FI1; -.
DR PDBsum; 4ATV; -.
DR PDBsum; 4AU5; -.
DR AlphaFoldDB; P13738; -.
DR SMR; P13738; -.
DR BioGRID; 4259720; 41.
DR DIP; DIP-10335N; -.
DR STRING; 511145.b0019; -.
DR BindingDB; P13738; -.
DR ChEMBL; CHEMBL5478; -.
DR TCDB; 2.A.33.1.1; the nhaa na(+):h(+) antiporter (nhaa) family.
DR jPOST; P13738; -.
DR PaxDb; P13738; -.
DR PRIDE; P13738; -.
DR EnsemblBacteria; AAC73130; AAC73130; b0019.
DR EnsemblBacteria; BAB96592; BAB96592; BAB96592.
DR GeneID; 944758; -.
DR KEGG; ecj:JW0018; -.
DR KEGG; eco:b0019; -.
DR PATRIC; fig|1411691.4.peg.2265; -.
DR EchoBASE; EB0646; -.
DR eggNOG; COG3004; Bacteria.
DR HOGENOM; CLU_015803_1_0_6; -.
DR InParanoid; P13738; -.
DR OMA; MGLMLRC; -.
DR PhylomeDB; P13738; -.
DR BioCyc; EcoCyc:NHAA-MON; -.
DR BioCyc; MetaCyc:NHAA-MON; -.
DR SABIO-RK; P13738; -.
DR EvolutionaryTrace; P13738; -.
DR PHI-base; PHI:3266; -.
DR PRO; PR:P13738; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005887; C:integral component of plasma membrane; IDA:EcoCyc.
DR GO; GO:0005886; C:plasma membrane; IDA:EcoCyc.
DR GO; GO:1901612; F:cardiolipin binding; IDA:EcoCyc.
DR GO; GO:0015385; F:sodium:proton antiporter activity; IDA:EcoCyc.
DR GO; GO:0051453; P:regulation of intracellular pH; TAS:EcoCyc.
DR GO; GO:0010446; P:response to alkaline pH; IDA:EcoCyc.
DR GO; GO:0009651; P:response to salt stress; IMP:EcoCyc.
DR Gene3D; 1.20.1530.10; -; 1.
DR HAMAP; MF_01844; NhaA; 1.
DR InterPro; IPR023171; Na/H_antiporter_dom_sf.
DR InterPro; IPR004670; NhaA.
DR PANTHER; PTHR30341; PTHR30341; 1.
DR Pfam; PF06965; Na_H_antiport_1; 1.
DR TIGRFAMs; TIGR00773; NhaA; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antiport; Cell inner membrane; Cell membrane;
KW Direct protein sequencing; Ion transport; Membrane; Reference proteome;
KW Sodium; Sodium transport; Transmembrane; Transmembrane helix; Transport.
FT CHAIN 1..388
FT /note="Na(+)/H(+) antiporter NhaA"
FT /id="PRO_0000052410"
FT TOPO_DOM 1..11
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 12..31
FT /note="Helical; Name=1"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 32..58
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 59..80
FT /note="Helical; Name=2"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 81..96
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 97..116
FT /note="Helical; Name=3"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 117..122
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 123..130
FT /note="Helical; Name=4"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 131..154
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 155..176
FT /note="Helical; Name=5"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 177..180
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 181..200
FT /note="Helical; Name=6"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 201..204
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 205..222
FT /note="Helical; Name=7"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 223
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 224..236
FT /note="Helical; Name=8"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 237..253
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 254..272
FT /note="Helical; Name=9"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 273..286
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 287..310
FT /note="Helical; Name=10"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 311..339
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 340..350
FT /note="Helical; Name=11"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 351..357
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TRANSMEM 358..380
FT /note="Helical; Name=12"
FT /evidence="ECO:0000269|PubMed:25422503,
FT ECO:0007744|PDB:4ATV, ECO:0007744|PDB:4AU5"
FT TOPO_DOM 381..388
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:15919996,
FT ECO:0000269|PubMed:25422503, ECO:0007744|PDB:4ATV,
FT ECO:0007744|PDB:4AU5"
FT REGION 45..58
FT /note="Important for dimerization"
FT /evidence="ECO:0000305|PubMed:17635927,
FT ECO:0000305|PubMed:23836890"
FT SITE 133
FT /note="Important for stability, cation binding and
FT translocation"
FT /evidence="ECO:0000305|PubMed:15988517,
FT ECO:0000305|PubMed:22915592, ECO:0000305|PubMed:27021484,
FT ECO:0000305|PubMed:29410365, ECO:0000305|PubMed:7737413"
FT SITE 163
FT /note="Essential for cation binding and translocation"
FT /evidence="ECO:0000305|PubMed:15988517,
FT ECO:0000305|PubMed:19274728, ECO:0000305|PubMed:22915592,
FT ECO:0000305|PubMed:25422503, ECO:0000305|PubMed:27021484,
FT ECO:0000305|PubMed:27708266, ECO:0000305|PubMed:30409911,
FT ECO:0000305|PubMed:7737413"
FT SITE 164
FT /note="Essential for cation binding and translocation"
FT /evidence="ECO:0000305|PubMed:15988517,
FT ECO:0000305|PubMed:19274728, ECO:0000305|PubMed:22915592,
FT ECO:0000305|PubMed:25422503, ECO:0000305|PubMed:27021484,
FT ECO:0000305|PubMed:27708266, ECO:0000305|PubMed:7737413"
FT SITE 241
FT /note="Important for pH response"
FT /evidence="ECO:0000305|PubMed:10455127"
FT SITE 252
FT /note="Important for pH response"
FT /evidence="ECO:0000305|PubMed:14604993"
FT SITE 254
FT /note="Important for pH response"
FT /evidence="ECO:0000305|PubMed:10455127"
FT SITE 300
FT /note="Important for stability and activity"
FT /evidence="ECO:0000305|PubMed:15988517,
FT ECO:0000305|PubMed:22915592, ECO:0000305|PubMed:25422503,
FT ECO:0000305|PubMed:27708266, ECO:0000305|PubMed:28330875,
FT ECO:0000305|PubMed:30409911"
FT MUTAGEN 45..58
FT /note="Missing: Exists exclusively in a monomeric form.
FT Shows antiporter activity under routine stress conditions,
FT but is much less efficient than wild-type dimeric NhaA in
FT conferring growth resistance under extreme stress
FT conditions. Does not affect pH dependence."
FT /evidence="ECO:0000269|PubMed:17635927,
FT ECO:0000269|PubMed:23836890"
FT MUTAGEN 65
FT /note="D->N: Does not impair antiporter activity. Mutant
FT can still survive in high NaC1 or LiC1 medium."
FT /evidence="ECO:0000269|PubMed:7737413"
FT MUTAGEN 132
FT /note="T->C: Decreases both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Increases Km for Na(+) and Li(+)."
FT /evidence="ECO:0000269|PubMed:22915592"
FT MUTAGEN 133
FT /note="D->A: Decreases both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Increases Km for Na(+) and Li(+).
FT Decreases thermal stability."
FT /evidence="ECO:0000269|PubMed:29410365"
FT MUTAGEN 133
FT /note="D->C: Decreases both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Increases Km for Na(+) and Li(+). 5-
FT fold decrease in Li(+) binding affinity. Changes the
FT H(+)/Li(+) stoichiometry to 4."
FT /evidence="ECO:0000269|PubMed:22915592,
FT ECO:0000269|PubMed:27021484, ECO:0000269|PubMed:29410365"
FT MUTAGEN 133
FT /note="D->K: Decreases both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Increases Km for Na(+) and Li(+).
FT Decreases thermal stability."
FT /evidence="ECO:0000269|PubMed:29410365"
FT MUTAGEN 133
FT /note="D->N: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities under all pH conditions examined.
FT Abolishes ability to grow on high salt medium."
FT /evidence="ECO:0000269|PubMed:7737413"
FT MUTAGEN 163
FT /note="D->C: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Abolishes ability to grow on high
FT salt medium. Cannot bind Li(+)."
FT /evidence="ECO:0000269|PubMed:11258962,
FT ECO:0000269|PubMed:22915592"
FT MUTAGEN 163
FT /note="D->E: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Abolishes ability to grow on high
FT salt medium."
FT /evidence="ECO:0000269|PubMed:19274728"
FT MUTAGEN 163
FT /note="D->N: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities under all pH conditions examined.
FT Abolishes ability to grow on high salt medium. Still active
FT and electrogenic; when associated with Q-300."
FT /evidence="ECO:0000269|PubMed:19274728,
FT ECO:0000269|PubMed:30409911, ECO:0000269|PubMed:7737413"
FT MUTAGEN 164
FT /note="D->C: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities. Abolishes ability to grow on high
FT salt medium. Cannot bind Li(+)."
FT /evidence="ECO:0000269|PubMed:11258962,
FT ECO:0000269|PubMed:22915592"
FT MUTAGEN 164
FT /note="D->E: Is active with both Na(+) and Li(+), however
FT with very high apparent Km values with both substrates. Can
FT grow on high Na(+) at neutral pH, albeit at a slower growth
FT rate than the wild type strain."
FT /evidence="ECO:0000269|PubMed:19274728"
FT MUTAGEN 164
FT /note="D->N: Loss of both Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities under all pH conditions examined.
FT Abolishes ability to grow on high salt medium."
FT /evidence="ECO:0000269|PubMed:19274728,
FT ECO:0000269|PubMed:7737413"
FT MUTAGEN 167
FT /note="A->P: Shows reduced Na(+)/H(+) and Li(+)/H(+)
FT antiporter activities, and a stronger down-regulation in
FT the alkaline range for Na(+) import. Converts the
FT antagonistic binding of the wild-type protein into
FT synergistic Li(+)/H(+) binding."
FT /evidence="ECO:0000269|PubMed:23836890,
FT ECO:0000269|PubMed:27021484"
FT MUTAGEN 225
FT /note="H->A: Loss of antiporter activity. Abolishes ability
FT to grow at alkaline pH."
FT /evidence="ECO:0000269|PubMed:7592922"
FT MUTAGEN 225
FT /note="H->C,S: Slightly reduced antiporter activity. No
FT effect on pH sensitivity."
FT /evidence="ECO:0000269|PubMed:7592922"
FT MUTAGEN 225
FT /note="H->D: Shifts threshold for pH-sensitive
FT inactivation."
FT /evidence="ECO:0000269|PubMed:7592922"
FT MUTAGEN 225
FT /note="H->R: Shifts threshold for pH-sensitive
FT inactivation. The pH dependence of Na(+) import is shifted
FT by 1 pH unit to the acidic range compared with the wild
FT type."
FT /evidence="ECO:0000269|PubMed:11258962,
FT ECO:0000269|PubMed:23836890, ECO:0000269|PubMed:7592922"
FT MUTAGEN 241
FT /note="E->C: Affects pH sensitivity. Shows acidic shift in
FT its pH profile. Causes a shift in the pH profile toward
FT basic pH; when associated with C-254."
FT /evidence="ECO:0000269|PubMed:10455127"
FT MUTAGEN 242..253
FT /note="Missing: Lack of Na(+)/H(+) antiporter activity at
FT pH 7, but shows weak activity at pH 8.5."
FT /evidence="ECO:0000269|PubMed:10455127"
FT MUTAGEN 249
FT /note="K->KIEG: Affects the pH sensitivity. The pH profile
FT of the activity is shifted by about half a pH unit toward
FT acidic pH."
FT /evidence="ECO:0000269|PubMed:10455127"
FT MUTAGEN 252
FT /note="E->C: Increases drastically the Km for Na(+). The pH
FT profile of the activity is shifted by one pH unit toward
FT the alkaline range."
FT /evidence="ECO:0000269|PubMed:14604993"
FT MUTAGEN 254
FT /note="V->C: Affects the pH sensitivity. The pH profile of
FT the activity is shifted by about half a pH unit toward
FT acidic pH. Causes a shift in the pH profile toward basic
FT pH; when associated with C-241. Another study shows no
FT acidic shift of the pH profile but rather a moderate
FT alkaline shift in the reverse transport direction."
FT /evidence="ECO:0000269|PubMed:10455127,
FT ECO:0000269|PubMed:23836890"
FT MUTAGEN 282
FT /note="D->N: Does not impair antiporter activity. Mutant
FT can still survive in high NaC1 or LiC1 medium."
FT /evidence="ECO:0000269|PubMed:7737413"
FT MUTAGEN 300
FT /note="K->A: Loss of Na(+)/H(+) antiporter activity, but
FT retains 30% of Li(+)/H(+) antiporter activity. Shows a wild
FT type-like pH dependence and a similar apparent Na(+)
FT affinity. Shows higher affinity for Li(+)."
FT /evidence="ECO:0000269|PubMed:28330875"
FT MUTAGEN 300
FT /note="K->C: Increases Km for Na(+)."
FT /evidence="ECO:0000269|PubMed:28330875"
FT MUTAGEN 300
FT /note="K->H: Retains 52% of Na(+)/H(+) antiporter activity
FT and 88% of Li(+)/H(+) antiporter activity. Increases Km for
FT Na(+) and Li(+). Does not affect pH-sensitivity. Decreases
FT thermal stability."
FT /evidence="ECO:0000269|PubMed:22915592,
FT ECO:0000269|PubMed:28330875"
FT MUTAGEN 300
FT /note="K->L: Loss of Na(+)/H(+) antiporter activity."
FT /evidence="ECO:0000269|PubMed:28330875"
FT MUTAGEN 300
FT /note="K->Q: Does not affect growth with Na(+) at pH 7, but
FT shows a moderate reduction in growth with Li(+). Growth is
FT decreased by 3 orders of magnitude under high-Na(+)
FT conditions at pH 8.2. Does not affect Na(+)/H(+) and
FT Li(+)/H(+) antiporter activities, but shows an increased Km
FT for Li(+). Decreases thermal stability. Still active and
FT electrogenic; when associated with N-163."
FT /evidence="ECO:0000269|PubMed:30409911"
FT MUTAGEN 300
FT /note="K->R: Retains 36% of Na(+)/H(+) antiporter activity
FT and 93% of Li(+)/H(+) antiporter activity. Cannot grow on
FT high Na(+) at pH 8.3. Increases Km for Na(+) and Li(+). The
FT pH profile is shifted to the alkaline side by one pH unit.
FT Decreases thermal stability."
FT /evidence="ECO:0000269|PubMed:22915592,
FT ECO:0000269|PubMed:28330875"
FT MUTAGEN 338
FT /note="G->S: Loss of pH sensitivity."
FT /evidence="ECO:0000269|PubMed:11258962"
FT HELIX 13..30
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 32..34
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 35..41
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 45..50
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 53..58
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 59..84
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 86..88
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 91..93
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 95..104
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 105..108
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 109..112
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 122..125
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 127..130
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 134..142
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 146..148
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 150..152
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 153..174
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 181..199
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 206..218
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 219..221
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 223..236
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 241..243
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 247..261
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 263..271
FT /evidence="ECO:0007829|PDB:1ZCD"
FT STRAND 277..279
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 284..287
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 290..293
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 294..298
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 301..303
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 305..311
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 315..317
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 325..328
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 331..334
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 339..349
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 351..353
FT /evidence="ECO:0007829|PDB:4ATV"
FT HELIX 358..370
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 375..377
FT /evidence="ECO:0007829|PDB:1ZCD"
FT HELIX 378..380
FT /evidence="ECO:0007829|PDB:1ZCD"
FT TURN 381..383
FT /evidence="ECO:0007829|PDB:1ZCD"
SQ SEQUENCE 388 AA; 41356 MW; B508B1D2E5EE9130 CRC64;
MKHLHRFFSS DASGGIILII AAILAMIMAN SGATSGWYHD FLETPVQLRV GSLEINKNML
LWINDALMAV FFLLVGLEVK RELMQGSLAS LRQAAFPVIA AIGGMIVPAL LYLAFNYADP
ITREGWAIPA ATDIAFALGV LALLGSRVPL ALKIFLMALA IIDDLGAIII IALFYTNDLS
MASLGVAAVA IAVLAVLNLC GARRTGVYIL VGVVLWTAVL KSGVHATLAG VIVGFFIPLK
EKHGRSPAKR LEHVLHPWVA YLILPLFAFA NAGVSLQGVT LDGLTSILPL GIIAGLLIGK
PLGISLFCWL ALRLKLAHLP EGTTYQQIMV VGILCGIGFT MSIFIASLAF GSVDPELINW
AKLGILVGSI SSAVIGYSWL RVRLRPSV
