ISPG_ECOLI
ID ISPG_ECOLI Reviewed; 372 AA.
AC P62620; P27433; P76984; P76985;
DT 19-JUL-2004, integrated into UniProtKB/Swiss-Prot.
DT 19-JUL-2004, sequence version 1.
DT 03-AUG-2022, entry version 144.
DE RecName: Full=4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (flavodoxin) {ECO:0000255|HAMAP-Rule:MF_00159};
DE EC=1.17.7.3 {ECO:0000255|HAMAP-Rule:MF_00159, ECO:0000269|PubMed:12434382, ECO:0000269|PubMed:12571359, ECO:0000269|PubMed:16268586};
DE AltName: Full=1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase {ECO:0000255|HAMAP-Rule:MF_00159};
DE AltName: Full=Protein GcpE;
DE Short=Protein E;
GN Name=ispG {ECO:0000255|HAMAP-Rule:MF_00159}; Synonyms=gcpE;
GN OrderedLocusNames=b2515, JW2499;
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=1521767; DOI=10.1016/0378-1097(92)90604-m;
RA Baker J., Franklin D.B., Parker J.;
RT "Sequence and characterization of the gcpE gene of Escherichia coli.";
RL FEMS Microbiol. Lett. 73:175-180(1992).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND CHARACTERIZATION.
RX PubMed=11752431; DOI=10.1073/pnas.201399298;
RA Hecht S., Eisenreich W., Adam P., Amslinger S., Kis K., Bacher A.,
RA Arigoni D., Rohdich F.;
RT "Studies on the nonmevalonate pathway to terpenes: the role of the GcpE
RT (IspG) protein.";
RL Proc. Natl. Acad. Sci. U.S.A. 98:14837-14842(2001).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=9205837; DOI=10.1093/dnares/4.2.91;
RA Yamamoto Y., Aiba H., Baba T., Hayashi K., Inada T., Isono K., Itoh T.,
RA Kimura S., Kitagawa M., Makino K., Miki T., Mitsuhashi N., Mizobuchi K.,
RA Mori H., Nakade S., Nakamura Y., Nashimoto H., Oshima T., Oyama S.,
RA Saito N., Sampei G., Satoh Y., Sivasundaram S., Tagami H., Takahashi H.,
RA Takeda J., Takemoto K., Uehara K., Wada C., Yamagata S., Horiuchi T.;
RT "Construction of a contiguous 874-kb sequence of the Escherichia coli-K12
RT genome corresponding to 50.0-68.8 min on the linkage map and analysis of
RT its sequence features.";
RL DNA Res. 4:91-113(1997).
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 PRELIMINARY NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 343-372.
RX PubMed=2991272; DOI=10.1016/s0021-9258(17)39212-8;
RA Freedman R., Gibson B., Donovan D., Biemann K., Eisenbeis S.J., Parker J.,
RA Schimmel P.;
RT "Primary structure of histidine-tRNA synthetase and characterization of
RT hisS transcripts.";
RL J. Biol. Chem. 260:10063-10068(1985).
RN [7]
RP PATHWAY.
RC STRAIN=K12 / MC4100 / ATCC 35695 / DSM 6574;
RX PubMed=11163766; DOI=10.1016/s0014-5793(00)02420-0;
RA Campos N., Rodriguez-Concepcion M., Seemann M., Rohmer M., Boronat A.;
RT "Identification of gcpE as a novel gene of the 2-C-methyl-D-erythritol 4-
RT phosphate pathway for isoprenoid biosynthesis in Escherichia coli.";
RL FEBS Lett. 488:170-173(2001).
RN [8]
RP PATHWAY.
RX PubMed=11274098; DOI=10.1128/jb.183.8.2411-2416.2001;
RA Altincicek B., Kollas A.-K., Sanderbrand S., Wiesner J., Hintz M., Beck E.,
RA Jomaa H.;
RT "GcpE is involved in the 2-C-methyl-D-erythritol 4-phosphate pathway of
RT isoprenoid biosynthesis in Escherichia coli.";
RL J. Bacteriol. 183:2411-2416(2001).
RN [9]
RP CATALYTIC ACTIVITY, COFACTOR, AND REACTION MECHANISM.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=12434382;
RX DOI=10.1002/1521-3773(20021115)41:22<4337::aid-anie4337>3.0.co;2-k;
RA Seemann M., Tse Sum Bui B., Wolff M., Tritsch D., Campos N., Boronat A.,
RA Marquet A., Rohmer M.;
RT "Isoprenoid biosynthesis through the methylerythritol phosphate pathway:
RT the (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase (GcpE) is a [4Fe-
RT 4S] protein.";
RL Angew. Chem. Int. Ed. 41:4337-4339(2002).
RN [10]
RP CATALYTIC ACTIVITY.
RX PubMed=12571359; DOI=10.1073/pnas.0337742100;
RA Rohdich F., Zepeck F., Adam P., Hecht S., Kaiser J., Laupitz R.,
RA Graewert T., Amslinger S., Eisenreich W., Bacher A., Arigoni D.;
RT "The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: studies on
RT the mechanisms of the reactions catalyzed by IspG and IspH protein.";
RL Proc. Natl. Acad. Sci. U.S.A. 100:1586-1591(2003).
RN [11]
RP MUTAGENESIS OF GLY-322; GLY-328 AND ARG-335.
RX PubMed=12859972; DOI=10.1016/s0006-291x(03)01211-7;
RA Sauret-Gueeto S., Ramos-Valdivia A., Ibanez E., Boronat A.,
RA Rodriguez-Concepcion M.;
RT "Identification of lethal mutations in Escherichia coli genes encoding
RT enzymes of the methylerythritol phosphate pathway.";
RL Biochem. Biophys. Res. Commun. 307:408-415(2003).
RN [12]
RP FUNCTION, AND REQUIREMENT FOR FLAVODOXIN.
RX PubMed=15978585; DOI=10.1016/j.febslet.2005.05.047;
RA Puan K.J., Wang H., Dairi T., Kuzuyama T., Morita C.T.;
RT "fldA is an essential gene required in the 2-C-methyl-D-erythritol 4-
RT phosphate pathway for isoprenoid biosynthesis.";
RL FEBS Lett. 579:3802-3806(2005).
RN [13]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND PH DEPENDENCE.
RX PubMed=16268586; DOI=10.1021/jo0510787;
RA Zepeck F., Grawert T., Kaiser J., Schramek N., Eisenreich W., Bacher A.,
RA Rohdich F.;
RT "Biosynthesis of isoprenoids. purification and properties of IspG protein
RT from Escherichia coli.";
RL J. Org. Chem. 70:9168-9174(2005).
CC -!- FUNCTION: Converts 2C-methyl-D-erythritol 2,4-cyclodiphosphate (ME-
CC 2,4cPP) into 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate, using
CC flavodoxin as the reducing agent. {ECO:0000255|HAMAP-Rule:MF_00159,
CC ECO:0000269|PubMed:15978585, ECO:0000269|PubMed:16268586}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(2E)-4-hydroxy-3-methylbut-2-enyl diphosphate + 2 H(+) + H2O +
CC oxidized [flavodoxin] = 2-C-methyl-D-erythritol 2,4-cyclic
CC diphosphate + reduced [flavodoxin]; Xref=Rhea:RHEA:43604, Rhea:RHEA-
CC COMP:10622, Rhea:RHEA-COMP:10623, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57618, ChEBI:CHEBI:58210,
CC ChEBI:CHEBI:58483, ChEBI:CHEBI:128753; EC=1.17.7.3;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00159,
CC ECO:0000269|PubMed:12434382, ECO:0000269|PubMed:12571359,
CC ECO:0000269|PubMed:16268586};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00159,
CC ECO:0000269|PubMed:12434382, ECO:0000269|PubMed:16268586};
CC Note=Binds 1 [4Fe-4S] cluster. {ECO:0000255|HAMAP-Rule:MF_00159,
CC ECO:0000269|PubMed:12434382, ECO:0000269|PubMed:16268586};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 9. {ECO:0000269|PubMed:16268586};
CC -!- PATHWAY: Isoprenoid biosynthesis; isopentenyl diphosphate biosynthesis
CC via DXP pathway; isopentenyl diphosphate from 1-deoxy-D-xylulose 5-
CC phosphate: step 5/6. {ECO:0000255|HAMAP-Rule:MF_00159,
CC ECO:0000269|PubMed:11163766, ECO:0000269|PubMed:11274098}.
CC -!- INTERACTION:
CC P62620; P77188: ecpB; NbExp=2; IntAct=EBI-550338, EBI-1113651;
CC -!- SIMILARITY: Belongs to the IspG family. {ECO:0000255|HAMAP-
CC Rule:MF_00159}.
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DR EMBL; X64451; CAA45783.1; -; Genomic_DNA.
DR EMBL; AY033515; AAK53460.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75568.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA16402.1; -; Genomic_DNA.
DR EMBL; M11843; -; NOT_ANNOTATED_CDS; Genomic_DNA.
DR PIR; S23058; S23058.
DR RefSeq; NP_417010.1; NC_000913.3.
DR RefSeq; WP_000551807.1; NZ_STEB01000011.1.
DR AlphaFoldDB; P62620; -.
DR SMR; P62620; -.
DR BioGRID; 4260590; 345.
DR BioGRID; 851330; 1.
DR DIP; DIP-35832N; -.
DR IntAct; P62620; 10.
DR STRING; 511145.b2515; -.
DR jPOST; P62620; -.
DR PaxDb; P62620; -.
DR PRIDE; P62620; -.
DR EnsemblBacteria; AAC75568; AAC75568; b2515.
DR EnsemblBacteria; BAA16402; BAA16402; BAA16402.
DR GeneID; 60903826; -.
DR GeneID; 67416899; -.
DR GeneID; 946991; -.
DR KEGG; ecj:JW2499; -.
DR KEGG; eco:b2515; -.
DR PATRIC; fig|1411691.4.peg.4221; -.
DR EchoBASE; EB0365; -.
DR eggNOG; COG0821; Bacteria.
DR InParanoid; P62620; -.
DR OMA; PTCGRTQ; -.
DR PhylomeDB; P62620; -.
DR BioCyc; EcoCyc:EG10370-MON; -.
DR BioCyc; MetaCyc:EG10370-MON; -.
DR BRENDA; 1.17.7.3; 2026.
DR UniPathway; UPA00056; UER00096.
DR PRO; PR:P62620; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:EcoCyc.
DR GO; GO:0046429; F:4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase activity; IDA:EcoCyc.
DR GO; GO:0005506; F:iron ion binding; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IDA:EcoCyc.
DR GO; GO:0019288; P:isopentenyl diphosphate biosynthetic process, methylerythritol 4-phosphate pathway; IMP:EcoCyc.
DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniRule.
DR Gene3D; 3.20.20.20; -; 1.
DR Gene3D; 3.30.413.10; -; 1.
DR HAMAP; MF_00159; IspG; 1.
DR InterPro; IPR011005; Dihydropteroate_synth-like.
DR InterPro; IPR016425; IspG_bac.
DR InterPro; IPR004588; IspG_bac-typ.
DR InterPro; IPR045854; NO2/SO3_Rdtase_4Fe4S_sf.
DR PANTHER; PTHR30454; PTHR30454; 1.
DR Pfam; PF04551; GcpE; 1.
DR PIRSF; PIRSF004640; IspG; 1.
DR SUPFAM; SSF51717; SSF51717; 1.
DR SUPFAM; SSF56014; SSF56014; 1.
DR TIGRFAMs; TIGR00612; ispG_gcpE; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Iron; Iron-sulfur; Isoprene biosynthesis; Metal-binding;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..372
FT /note="4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase
FT (flavodoxin)"
FT /id="PRO_0000190572"
FT BINDING 270
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00159"
FT BINDING 273
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00159"
FT BINDING 305
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00159"
FT BINDING 312
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00159"
FT MUTAGEN 322
FT /note="G->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12859972"
FT MUTAGEN 328
FT /note="G->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12859972"
FT MUTAGEN 335
FT /note="R->C: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12859972"
SQ SEQUENCE 372 AA; 40684 MW; 3666FD83D5CD4F9F CRC64;
MHNQAPIQRR KSTRIYVGNV PIGDGAPIAV QSMTNTRTTD VEATVNQIKA LERVGADIVR
VSVPTMDAAE AFKLIKQQVN VPLVADIHFD YRIALKVAEY GVDCLRINPG NIGNEERIRM
VVDCARDKNI PIRIGVNAGS LEKDLQEKYG EPTPQALLES AMRHVDHLDR LNFDQFKVSV
KASDVFLAVE SYRLLAKQID QPLHLGITEA GGARSGAVKS AIGLGLLLSE GIGDTLRVSL
AADPVEEIKV GFDILKSLRI RSRGINFIAC PTCSRQEFDV IGTVNALEQR LEDIITPMDV
SIIGCVVNGP GEALVSTLGV TGGNKKSGLY EDGVRKDRLD NNDMIDQLEA RIRAKASQLD
EARRIDVQQV EK
