PUTP_ECOLI
ID PUTP_ECOLI Reviewed; 502 AA.
AC P07117;
DT 01-APR-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-1988, sequence version 1.
DT 03-AUG-2022, entry version 177.
DE RecName: Full=Sodium/proline symporter {ECO:0000305};
DE AltName: Full=Proline carrier {ECO:0000303|PubMed:3902503};
DE AltName: Full=Proline permease;
DE AltName: Full=Propionate transporter {ECO:0000303|PubMed:17088549};
GN Name=putP; OrderedLocusNames=b1015, JW1001;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=3302614; DOI=10.1007/bf00330424;
RA Nakao T., Yamato I., Anraku Y.;
RT "Nucleotide sequence of putP, the proline carrier gene of Escherichia coli
RT K12.";
RL Mol. Gen. Genet. 208:70-75(1987).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=8905232; DOI=10.1093/dnares/3.3.137;
RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K.,
RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S.,
RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H.,
RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G.,
RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M.,
RA Horiuchi T.;
RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 12.7-28.0 min region on the linkage map.";
RL DNA Res. 3:137-155(1996).
RN [3]
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 [4]
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 [5]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-489.
RC STRAIN=EC14;
RX PubMed=1400239; DOI=10.1128/jb.174.21.6886-6895.1992;
RA Nelson K., Selander R.K.;
RT "Evolutionary genetics of the proline permease gene (putP) and the control
RT region of the proline utilization operon in populations of Salmonella and
RT Escherichia coli.";
RL J. Bacteriol. 174:6886-6895(1992).
RN [6]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-43.
RC STRAIN=K12;
RX PubMed=3325781; DOI=10.1007/bf00325707;
RA Nakao T., Yamato I., Anraku Y.;
RT "Nucleotide sequence of putC, the regulatory region for the put regulon of
RT Escherichia coli K12.";
RL Mol. Gen. Genet. 210:364-368(1987).
RN [7]
RP PROTEIN SEQUENCE OF 1-6, FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP SODIUM/PROLINE BINDING SITE, AND MUTAGENESIS OF CYS-281; CYS-344 AND
RP CYS-349.
RX PubMed=1567896; DOI=10.1016/0005-2736(92)90162-f;
RA Hanada K., Yoshida T., Yamato I., Anraku Y.;
RT "Sodium ion and proline binding sites in the Na+/proline symport carrier of
RT Escherichia coli.";
RL Biochim. Biophys. Acta 1105:61-66(1992).
RN [8]
RP SUBCELLULAR LOCATION.
RC STRAIN=K12;
RX PubMed=3902503; DOI=10.1016/0014-5793(85)80024-7;
RA Hanada K., Yamato I., Anraku Y.;
RT "Identification of proline carrier in Escherichia coli K-12.";
RL FEBS Lett. 191:278-282(1985).
RN [9]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND SUBCELLULAR LOCATION.
RC STRAIN=K12;
RX PubMed=3512540; DOI=10.1016/s0021-9258(17)35829-5;
RA Chen C.C., Wilson T.H.;
RT "Solubilization and functional reconstitution of the proline transport
RT system of Escherichia coli.";
RL J. Biol. Chem. 261:2599-2604(1986).
RN [10]
RP SUBCELLULAR LOCATION, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12;
RX PubMed=3007935; DOI=10.1007/bf00330513;
RA Mogi T., Yamamoto H., Nakao T., Yamato I., Anraku Y.;
RT "Genetic and physical characterization of putP, the proline carrier gene of
RT Escherichia coli K12.";
RL Mol. Gen. Genet. 202:35-41(1986).
RN [11]
RP ACTIVITY REGULATION, AND MUTAGENESIS OF CYS-281; CYS-344 AND CYS-349.
RX PubMed=3053687; DOI=10.1016/s0021-9258(18)37556-2;
RA Yamato I., Anraku Y.;
RT "Site-specific alteration of cysteine 281, cysteine 344, and cysteine 349
RT in the proline carrier of Escherichia coli.";
RL J. Biol. Chem. 263:16055-16057(1988).
RN [12]
RP MUTAGENESIS OF ARG-257.
RX PubMed=3053649; DOI=10.1128/jb.170.11.5185-5191.1988;
RA Ohsawa M., Mogi T., Yamamoto H., Yamato I., Anraku Y.;
RT "Proline carrier mutant of Escherichia coli K-12 with altered cation
RT sensitivity of substrate-binding activity: cloning, biochemical
RT characterization, and identification of the mutation.";
RL J. Bacteriol. 170:5185-5191(1988).
RN [13]
RP MUTAGENESIS OF ARG-376.
RX PubMed=8119910; DOI=10.1016/s0021-9258(17)37520-8;
RA Yamato I., Kotani M., Oka Y., Anraku Y.;
RT "Site-specific alteration of arginine 376, the unique positively charged
RT amino acid residue in the mid-membrane-spanning regions of the proline
RT carrier of Escherichia coli.";
RL J. Biol. Chem. 269:5720-5724(1994).
RN [14]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=9693004; DOI=10.1021/bi980684b;
RA Jung H., Tebbe S., Schmid R., Jung K.;
RT "Unidirectional reconstitution and characterization of purified Na+/proline
RT transporter of Escherichia coli.";
RL Biochemistry 37:11083-11088(1998).
RN [15]
RP SUBCELLULAR LOCATION, AND TOPOLOGY.
RX PubMed=9756872; DOI=10.1074/jbc.273.41.26400;
RA Jung H., Ruebenhagen R., Tebbe S., Leifker K., Tholema N., Quick M.,
RA Schmid R.;
RT "Topology of the Na+/proline transporter of Escherichia coli.";
RL J. Biol. Chem. 273:26400-26407(1998).
RN [16]
RP SUBUNIT, AND SUBCELLULAR LOCATION.
RC STRAIN=BL21-DE3;
RX PubMed=16079137; DOI=10.1074/jbc.m506479200;
RA Stenberg F., Chovanec P., Maslen S.L., Robinson C.V., Ilag L.,
RA von Heijne G., Daley D.O.;
RT "Protein complexes of the Escherichia coli cell envelope.";
RL J. Biol. Chem. 280:34409-34419(2005).
RN [17]
RP 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 [18]
RP FUNCTION AS A PROPIONATE TRANSPORTER, ROLE IN PROPIONATE UTILIZATION,
RP DISRUPTION PHENOTYPE, AND INDUCTION.
RX PubMed=17088549; DOI=10.1073/pnas.0603364103;
RA Reed J.L., Patel T.R., Chen K.H., Joyce A.R., Applebee M.K., Herring C.D.,
RA Bui O.T., Knight E.M., Fong S.S., Palsson B.O.;
RT "Systems approach to refining genome annotation.";
RL Proc. Natl. Acad. Sci. U.S.A. 103:17480-17484(2006).
CC -!- FUNCTION: Catalyzes the sodium-dependent uptake of extracellular L-
CC proline (PubMed:3512540, PubMed:1567896, PubMed:9693004). This protein
CC is also capable of using lithium as the transport cation
CC (PubMed:1567896, PubMed:9693004). Also catalyzes the uptake of
CC propionate (PubMed:17088549). {ECO:0000269|PubMed:1567896,
CC ECO:0000269|PubMed:17088549, ECO:0000269|PubMed:3512540,
CC ECO:0000269|PubMed:9693004}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-proline(in) + Na(+)(in) = L-proline(out) + Na(+)(out);
CC Xref=Rhea:RHEA:28967, ChEBI:CHEBI:29101, ChEBI:CHEBI:60039;
CC Evidence={ECO:0000269|PubMed:1567896, ECO:0000269|PubMed:3512540,
CC ECO:0000269|PubMed:9693004};
CC -!- ACTIVITY REGULATION: Activity is stimulated by phosphatidylethanolamine
CC and phosphatidylglycerol, but not by phosphatidylcholine and
CC cardiolipin (PubMed:3512540). Proline uptake is inhibited by the
CC sulfhydryl reagent N-ethylmaleimide (NEM) (PubMed:3053687,
CC PubMed:1567896). Proline, in the presence of Na(+) or Li(+), protects
CC the carrier functions from NEM-inactivation (PubMed:1567896).
CC {ECO:0000269|PubMed:1567896, ECO:0000269|PubMed:3053687,
CC ECO:0000269|PubMed:3512540}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.5 uM for L-proline {ECO:0000269|PubMed:3512540};
CC KM=2 uM for L-proline (for Na(+)-driven transport)
CC {ECO:0000269|PubMed:9693004};
CC KM=31 uM for Na(+) (in intact cells) {ECO:0000269|PubMed:9693004};
CC KM=730 uM for Na(+) (for reconstituted PutP in proteoliposomes)
CC {ECO:0000269|PubMed:9693004};
CC Vmax=16 nmol/min/mg enzyme {ECO:0000269|PubMed:3512540};
CC Vmax=1130 nmol/min/mg enzyme (for Na(+)-driven transport)
CC {ECO:0000269|PubMed:9693004};
CC pH dependence:
CC Optimum pH is 7.0. {ECO:0000269|PubMed:3512540};
CC -!- SUBUNIT: Has been isolated from inner membrane preparations as a
CC homodimer. {ECO:0000269|PubMed:16079137}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane {ECO:0000269|PubMed:16079137,
CC ECO:0000269|PubMed:3007935, ECO:0000269|PubMed:3512540,
CC ECO:0000269|PubMed:3902503, ECO:0000269|PubMed:9756872}; Multi-pass
CC membrane protein {ECO:0000269|PubMed:16079137,
CC ECO:0000269|PubMed:9756872}.
CC -!- INDUCTION: Up-regulated by propionate. {ECO:0000269|PubMed:17088549}.
CC -!- DISRUPTION PHENOTYPE: Mutant is defective in proline transport
CC (PubMed:3007935). Shows reduced growth rate when grown on propionate as
CC sole carbon source (PubMed:17088549). {ECO:0000269|PubMed:17088549,
CC ECO:0000269|PubMed:3007935}.
CC -!- SIMILARITY: Belongs to the sodium:solute symporter (SSF) (TC 2.A.21)
CC family. {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; X05653; CAA29143.1; -; Genomic_DNA.
DR EMBL; U00096; AAC74100.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35793.1; -; Genomic_DNA.
DR EMBL; L01150; AAA24463.1; -; Genomic_DNA.
DR EMBL; L01151; AAA24464.1; -; Genomic_DNA.
DR EMBL; M35174; AAA24458.1; -; Genomic_DNA.
DR PIR; A30258; JGECPP.
DR RefSeq; NP_415535.1; NC_000913.3.
DR RefSeq; WP_001018479.1; NZ_SSZK01000002.1.
DR AlphaFoldDB; P07117; -.
DR SMR; P07117; -.
DR BioGRID; 4260051; 14.
DR BioGRID; 849976; 3.
DR IntAct; P07117; 4.
DR STRING; 511145.b1015; -.
DR TCDB; 2.A.21.2.1; the solute:sodium symporter (sss) family.
DR PaxDb; P07117; -.
DR PRIDE; P07117; -.
DR EnsemblBacteria; AAC74100; AAC74100; b1015.
DR EnsemblBacteria; BAA35793; BAA35793; BAA35793.
DR GeneID; 945602; -.
DR KEGG; ecj:JW1001; -.
DR KEGG; eco:b1015; -.
DR PATRIC; fig|1411691.4.peg.1254; -.
DR EchoBASE; EB0795; -.
DR eggNOG; COG0591; Bacteria.
DR HOGENOM; CLU_018808_15_2_6; -.
DR InParanoid; P07117; -.
DR OMA; CWTDLIQ; -.
DR PhylomeDB; P07117; -.
DR BioCyc; EcoCyc:PUTP-MON; -.
DR BioCyc; MetaCyc:PUTP-MON; -.
DR PRO; PR:P07117; -.
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:0005298; F:proline:sodium symporter activity; IDA:EcoCyc.
DR GO; GO:0031402; F:sodium ion binding; IEA:InterPro.
DR GO; GO:0015824; P:proline transport; IDA:EcoCyc.
DR GO; GO:0015912; P:short-chain fatty acid transport; IMP:EcoCyc.
DR CDD; cd11475; SLC5sbd_PutP; 1.
DR Gene3D; 1.20.1730.10; -; 1.
DR InterPro; IPR038377; Na/Glc_symporter_sf.
DR InterPro; IPR011851; Na/Pro_symporter.
DR InterPro; IPR001734; Na/solute_symporter.
DR InterPro; IPR018212; Na/solute_symporter_CS.
DR Pfam; PF00474; SSF; 1.
DR TIGRFAMs; TIGR02121; Na_Pro_sym; 1.
DR TIGRFAMs; TIGR00813; sss; 1.
DR PROSITE; PS00456; NA_SOLUT_SYMP_1; 1.
DR PROSITE; PS00457; NA_SOLUT_SYMP_2; 1.
DR PROSITE; PS50283; NA_SOLUT_SYMP_3; 1.
PE 1: Evidence at protein level;
KW Amino-acid transport; Cell inner membrane; Cell membrane;
KW Direct protein sequencing; Ion transport; Membrane; Reference proteome;
KW Sodium; Sodium transport; Symport; Transmembrane; Transmembrane helix;
KW Transport.
FT CHAIN 1..502
FT /note="Sodium/proline symporter"
FT /id="PRO_0000105398"
FT TOPO_DOM 1..5
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 6..26
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 27..41
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 42..62
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 63..67
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 68..88
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 89..126
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 127..147
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 148..162
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 163..183
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 184..192
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 193..213
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 214..234
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 235..255
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 256..275
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 276..296
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 297..319
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 320..340
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 341..370
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 371..391
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 392..397
FT /note="Periplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 398..418
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 419..427
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:9756872"
FT TRANSMEM 428..448
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TRANSMEM 449..469
FT /note="Helical"
FT /evidence="ECO:0000255"
FT TOPO_DOM 470..502
FT /note="Cytoplasmic"
FT /evidence="ECO:0000269|PubMed:15919996,
FT ECO:0000269|PubMed:9756872"
FT REGION 27..66
FT /note="Hydrophilic"
FT REGION 88..124
FT /note="Hydrophilic"
FT REGION 151..162
FT /note="Hydrophilic"
FT REGION 185..189
FT /note="Hydrophilic"
FT REGION 214..231
FT /note="Hydrophilic"
FT REGION 249..274
FT /note="Hydrophilic"
FT REGION 296..319
FT /note="Hydrophilic"
FT REGION 341..370
FT /note="Hydrophilic"
FT REGION 392..397
FT /note="Hydrophilic"
FT REGION 424..430
FT /note="Hydrophilic"
FT REGION 446..448
FT /note="Hydrophilic"
FT REGION 476..502
FT /note="Hydrophilic"
FT SITE 344
FT /note="Involved in high-affinity binding for sodium and
FT proline"
FT /evidence="ECO:0000305|PubMed:1567896"
FT MUTAGEN 257
FT /note="R->C: Sodium-independent binding affinity for
FT proline."
FT /evidence="ECO:0000269|PubMed:3053649"
FT MUTAGEN 281
FT /note="C->S: Does not affect proline uptake activity.
FT Confers resistance to N-ethylmaleimide. Na(+)-dependent
FT proline binding activity is similar to wild-type carrier."
FT /evidence="ECO:0000269|PubMed:1567896,
FT ECO:0000269|PubMed:3053687"
FT MUTAGEN 344
FT /note="C->S: Small decrease in proline uptake activity.
FT Confers resistance to N-ethylmaleimide. Exhibits low Na(+)-
FT dependent proline binding."
FT /evidence="ECO:0000269|PubMed:1567896,
FT ECO:0000269|PubMed:3053687"
FT MUTAGEN 349
FT /note="C->S: Does not affect proline uptake activity.
FT Sensitive to N-ethylmaleimide. Na(+)-dependent proline
FT binding activity is similar to wild-type carrier."
FT /evidence="ECO:0000269|PubMed:1567896,
FT ECO:0000269|PubMed:3053687"
FT MUTAGEN 376
FT /note="R->E,Q: No change in activity."
FT /evidence="ECO:0000269|PubMed:8119910"
FT MUTAGEN 376
FT /note="R->K: Loss of activity."
FT /evidence="ECO:0000269|PubMed:8119910"
SQ SEQUENCE 502 AA; 54344 MW; 37AE923C33186356 CRC64;
MAISTPMLVT FCVYIFGMIL IGFIAWRSTK NFDDYILGGR SLGPFVTALS AGASDMSGWL
LMGLPGAVFL SGISESWIAI GLTLGAWINW KLVAGRLRVH TEYNNNALTL PDYFTGRFED
KSRILRIISA LVILLFFTIY CASGIVAGAR LFESTFGMSY ETALWAGAAA TILYTFIGGF
LAVSWTDTVQ ASLMIFALIL TPVIVIISVG GFGDSLEVIK QKSIENVDML KGLNFVAIIS
LMGWGLGYFG QPHILARFMA ADSHHSIVHA RRISMTWMIL CLAGAVAVGF FGIAYFNDHP
ALAGAVNQNA ERVFIELAQI LFNPWIAGIL LSAILAAVMS TLSCQLLVCS SAITEDLYKA
FLRKHASQKE LVWVGRVMVL VVALVAIALA ANPENRVLGL VSYAWAGFGA AFGPVVLFSV
MWSRMTRNGA LAGMIIGALT VIVWKQFGWL GLYEIIPGFI FGSIGIVVFS LLGKAPSAAM
QKRFAEADAH YHSAPPSRLQ ES
